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Sample records for swab samples electronic

  1. Swab sample transfer for point-of-care diagnostics: characterization of swab types and manual agitation methods.

    Directory of Open Access Journals (Sweden)

    Nuttada Panpradist

    Full Text Available BACKGROUND: The global need for disease detection and control has increased effort to engineer point-of-care (POC tests that are simple, robust, affordable, and non-instrumented. In many POC tests, sample collection involves swabbing the site (e.g., nose, skin, agitating the swab in a fluid to release the sample, and transferring the fluid to a device for analysis. Poor performance in sample transfer can reduce sensitivity and reproducibility. METHODS: In this study, we compared bacterial release efficiency of seven swab types using manual-agitation methods typical of POC devices. Transfer efficiency was measured using quantitative PCR (qPCR for Staphylococcus aureus under conditions representing a range of sampling scenarios: 1 spiking low-volume samples onto the swab, 2 submerging the swab in excess-volume samples, and 3 swabbing dried sample from a surface. RESULTS: Excess-volume samples gave the expected recovery for most swabs (based on tip fluid capacity; a polyurethane swab showed enhanced recovery, suggesting an ability to accumulate organisms during sampling. Dry samples led to recovery of ∼20-30% for all swabs tested, suggesting that swab structure and volume is less important when organisms are applied to the outer swab surface. Low-volume samples led to the widest range of transfer efficiencies between swab types. Rayon swabs (63 µL capacity performed well for excess-volume samples, but showed poor recovery for low-volume samples. Nylon (100 µL and polyester swabs (27 µL showed intermediate recovery for low-volume and excess-volume samples. Polyurethane swabs (16 µL showed excellent recovery for all sample types. This work demonstrates that swab transfer efficiency can be affected by swab material, structure, and fluid capacity and details of the sample. Results and quantitative analysis methods from this study will assist POC assay developers in selecting appropriate swab types and transfer methods.

  2. Comparison of air samples, nasal swabs, ear-skin swabs and environmental dust samples for detection of Methicillin Resistant Staphylococcus aureus (MRSA) in pig herds

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Vigre, Håkan; Cavaco, Lina

    2014-01-01

    To identify a cost-effective and practical method for detection of methicillin-resistant Staphylococcus aureus (MRSA) in pig herds, the relative sensitivity of four sample types: nasal swabs, ear-skin (skin behind the ears) swabs, environmental dust swabs and air was compared. Moreover, dependency......-herd prevalence ⩾25%]. The results indicate that taking swabs of skin behind the ears (ten pools of five) was even more sensitive than taking nasal swabs (ten pools of five) at the herd level and detected significantly more positive samples. spa types t011, t034 and t4208 were observed. In conclusion, MRSA...... detection by air sampling is easy to perform, reduces costs and analytical time compared to existing methods, and is recommended for initial testing of herds. Ear-skin swab sampling may be more sensitive for MRSA detection than air sampling or nasal swab sampling....

  3. PE-Swab Direct STR Amplification of Forensic Touch DNA Samples.

    Science.gov (United States)

    Liu, Jason Y

    2015-05-01

    The PE-Swab direct STR amplification workflow was developed to process low-level "touch DNA" samples. In this workflow, a forensic sample is first collected on a 4-mm PE-Swab (a novel sample collection device); two 2-mm punches containing collected samples are then generated from the PE-Swab and directly amplified for STR typing. Compared to the conventional STR workflow, which involves DNA extraction, purification, and elution volume reduction, the PE-Swab direct STR amplification workflow does not require sample preparation and takes DNA loss due to sample preparation, the PE-Swab workflow is more sensitive than the conventional STR workflow. The average peak height per sample obtained by the PE-swab workflow is 3 times higher than that from the conventional workflow with both low-level single source and two-contributor mixture samples tested in this study. © 2015 American Academy of Forensic Sciences.

  4. Experimental comparison of excision and swabbing microbiological sampling methods for carcasses.

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    Pepperell, Richard; Reid, Carol-Ann; Solano, Silvia Nicolau; Hutchison, Michael L; Walters, Lisa D; Johnston, Alexander M; Buncic, Sava

    2005-10-01

    Bovine sides, ovine carcasses, and porcine carcasses were individually inoculated by dipping in various suspensions of a marker organism (Escherichia coli K-12 or Pseudomonas fluorescens), alone or in combination with two meat-derived bacterial strains, and were sampled by two standard methods: cotton wet-dry swabbing and excision. The samples were examined for bacterial counts on plate count agar (PCA plate counts) and on violet red brilliant green agar (VRBGA plate counts) by standard International Organization for Standardization methods. Average bacterial recoveries by swabbing, expressed as a percentage of the appropriate recoveries achieved by excision, varied widely (2 to 100%). Several factors that potentially contributed to relatively low and highly variable bacterial recoveries obtained by swabbing were investigated in separate experiments. Neither the difference in size of the swabbed area (10, 50, or 100 cm2 on beef carcasses) nor the difference in time of swabbing (20 or 60 min after inoculation of pig carcasses) had a significant effect on the swabbing recoveries of the marker organism used. In an experiment with swabs preinoculated with the marker organism and then used for carcass swabbing, on average, 12% of total bacterial load was transferred inversely (i.e., from the swab to the carcass during the standard swabbing procedure). In another experiment, on average, 14% of total bacterial load was not released from the swab into the diluent during standard swab homogenization. Use of custom-made swabs with abrasive butts, around which metal pieces of pan scourers were wound, markedly increased PCA plate count recoveries from noninoculated lamb carcasses at commercial abattoirs compared with cotton swabs. In spite of the observed inferiority of the cotton wet-dry swabbing method compared with the excision method for bacterial recovery, the former is clearly preferred by the meat industry because it does not damage the carcass. Therefore, further large

  5. A simplified field protocol for genetic sampling of birds using buccal swabs

    Science.gov (United States)

    Vilstrup, Julia T.; Mullins, Thomas D.; Miller, Mark P.; McDearman, Will; Walters, Jeffrey R.; Haig, Susan M.

    2018-01-01

    DNA sampling is an essential prerequisite for conducting population genetic studies. For many years, blood sampling has been the preferred method for obtaining DNA in birds because of their nucleated red blood cells. Nonetheless, use of buccal swabs has been gaining favor because they are less invasive yet still yield adequate amounts of DNA for amplifying mitochondrial and nuclear markers; however, buccal swab protocols often include steps (e.g., extended air-drying and storage under frozen conditions) not easily adapted to field settings. Furthermore, commercial extraction kits and swabs for buccal sampling can be expensive for large population studies. We therefore developed an efficient, cost-effective, and field-friendly protocol for sampling wild birds after comparing DNA yield among 3 inexpensive buccal swab types (2 with foam tips and 1 with a cotton tip). Extraction and amplification success was high (100% and 97.2% respectively) using inexpensive generic swabs. We found foam-tipped swabs provided higher DNA yields than cotton-tipped swabs. We further determined that omitting a drying step and storing swabs in Longmire buffer increased efficiency in the field while still yielding sufficient amounts of DNA for detailed population genetic studies using mitochondrial and nuclear markers. This new field protocol allows time- and cost-effective DNA sampling of juveniles or small-bodied birds for which drawing blood may cause excessive stress to birds and technicians alike.

  6. Broth versus solid agar culture of swab samples of cadaveric allograft musculoskeletal tissue.

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    Varettas, Kerry

    2013-12-01

    As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006-2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO2, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.

  7. New technique to take samples from environmental surfaces using flocked nylon swabs.

    Science.gov (United States)

    Hedin, G; Rynbäck, J; Loré, B

    2010-08-01

    Environmental surfaces near infected and/or colonised patients in hospitals are commonly contaminated with potentially pathogenic micro-organisms. At present, however, there is no standardised method for taking samples from surfaces in order to perform quantitative cultures. Usually contact plates or swabs are used, but these methods may give different results. The recovery rate of traditional swabbing, e.g. cotton or rayon, is poor. With a new type of swab utilising flocked nylon, the recovery may be enhanced up to three times compared with a rayon swab. In this study, we inoculated reference strains of Staphylococcus aureus and Enterococcus hirae onto a bedside table and took samples 1h later when inocula were dry. Sequential samples were taken from the same surface. A new sampling technique using two sequential nylon swabs for each sample was validated. The efficiency of the sampling, percentage recovery of the inoculum and the variation of culture results obtained from repeated experiments are described. Enhanced efficiency and higher recovery of inoculum were demonstrated using two sequential flocked nylon swabs for sampling. Copyright 2010 The Hospital Infection Society. Published by Elsevier Ltd. All rights reserved.

  8. Use of swabs for sampling epithelial cells for molecular genetics analyses in Enteroctopus

    Science.gov (United States)

    Hollenback, Nathan; Scheel, David; Gravley, Meg C.; Sage, George K.; Toussaint, Rebecca K.; Talbot, Sandra

    2017-01-01

    We evaluated the efficacy of using swabs to collect cells from the epidermis of octopus as a non-invasive DNA source for classical genetic studies, and demonstrated value of the technique by incorporating it into an effort to determine, within a day, the lineage of captured, live Enteroctopus (E. dofleini or a cryptic lineage). The cryptic lineage was targeted for captive behavioral and morphological studies, while once genetically identified, the non-target lineage could be more rapidly released back to the wild. We used commercially available sterile foamtipped swabs and a high-salt preservation buffer to collect and store paired swab and muscle (arm tip) tissue sampled from live Enteroctopus collected from Prince William Sound, Alaska. We performed a one-day extraction of DNA from epithelial swab samples and amplification of two diagnostic microsatellite loci to determine the lineage of each of the 21 individuals. Following this rapid lineage assessment, which allowed us to release non-target individuals within a day of laboratory work, we compared paired swab and muscle tissue samples from each individual to assess quantity of DNA yields and consistency of genotyping results, followed by assessment of locus-by-locus reliability of DNA extracts from swabs. Epithelial swabs yielded, on average, lower quantities of DNA (170.32 ± 74.72 (SD) ng/μL) relative to DNA obtained from tissues collected using invasive or destructive techniques (310.95 ± 147.37 (SD) ng/μL. We observed some decrease in yields of DNA from extractions of swab samples conducted 19 and 31 months after initial extractions when samples were stored at room temperature in lysis buffer. All extractions yielded quantities of DNA sufficient to amplify and score all loci, which included fragment data from 10 microsatellite loci (nine polymorphic loci and monomorphic locus EdoμA106), and nucleotide sequence data from a 528 base pair portion of the nuclear octopine dehydrogenase gene. All results

  9. Surface-sampling and analysis of TATP by swabbing and gas chromatography/mass spectrometry.

    Science.gov (United States)

    Romolo, Francesco Saverio; Cassioli, Luigi; Grossi, Silvana; Cinelli, Giuseppe; Russo, Mario Vincenzo

    2013-01-10

    The method of sample recovery for trace detection and identification of explosives plays a critical role in several criminal investigations. After bombing, there can be difficulties in sending big objects to a laboratory for analysis. Traces can also be searched for on large surfaces, on hands of suspects or on surfaces where the explosive was placed during preparatory phases (e.g. places where an IED was assembled, vehicles used for transportation, etc.). In this work, triacetone triperoxide (TATP) was synthesized from commercial precursors following reported methods. Several portions of about 6mg of TATP were then spread on different surfaces (e.g. floors, tables, etc.) or used in handling tests. Three different swabbing systems were used: a commercial swab, pre-wetted with propan-2-ol (isopropanol) and water (7:3), dry paper swabs, and cotton swabs wetted with propan-2-ol. Paper and commercial swabs were also used to sample a metal plate, where a small charge of about 4g of TATP was detonated. Swabs were sealed in small glass jars with screw caps and Parafilm(®) M and sent to the laboratory for analysis. Swabs were extracted and analysed several weeks later by gas chromatography/mass spectrometry. All the three systems gave positive results, but wetted swabs collected higher amounts of TATP. The developed procedure showed its suitability for use in real cases, allowing TATP detection in several simulations, including a situation in which people wash their hands after handling the explosive. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  10. Post-coital vaginal sampling with nylon flocked swabs improves DNA typing.

    Science.gov (United States)

    Benschop, Corina C G; Wiebosch, Danielle C; Kloosterman, Ate D; Sijen, Titia

    2010-02-01

    In the examination of sexual assault cases, DNA typing of vaginal samples mostly occurs after differential DNA extraction. Notwithstanding the differential extraction method, the DNA profiles from the seminal fraction often show the male alleles at low-level in combination with female alleles. This unfavorable ratio male to female DNA is due to a limited amount of sperm cells and an overwhelming quantity of female cells. In this study, we compared standard cotton and nylon flocked swabs for post-coital vaginal sampling. Twelve couples donated 88 vaginal swabs - 44 cotton, 44 nylon flocked - which were taken with a time since intercourse (TSI) up to 84 h. These vaginal swabs were sorted into categories on the basis of the TSI and submitted to (1) microscopic examination for the presence of male cells, (2) presumptive tests for the detection of seminal fluid and (3) DNA typing. Cellular elution was found to be 6-fold more efficient from the nylon flocked swabs. This makes microscopic analysis less time consuming as the higher cell yield and better cell morphology simplify detection of male cells. Both swab types reveal similar results regarding presumptive tests and male DNA typing. Positive presumptive tests (RSID-semen and PSA) were obtained up to 60 h TSI and male autosomal profiles up to 72 h TSI. Interestingly, over 50% of the samples negative for both presumptive tests resulted in informative male STR profiles. After differential extraction, less DNA was left on the nylon flocked swabs and more male DNA was isolated. Our results imply that the use of nylon flocked swabs for vaginal sampling will improve microscopic analysis and DNA typing in the medical forensic investigation of sexual assault cases.

  11. High-throughput sequencing of forensic genetic samples using punches of FTA cards with buccal swabs

    DEFF Research Database (Denmark)

    Kampmann, Marie-Louise; Buchard, Anders; Børsting, Claus

    2016-01-01

    Here, we demonstrate that punches from buccal swab samples preserved on FTA cards can be used for high-throughput DNA sequencing, also known as massively parallel sequencing (MPS). We typed 44 reference samples with the HID-Ion AmpliSeq Identity Panel using washed 1.2 mm punches from FTA cards...... with buccal swabs and compared the results with those obtained with DNA extracted using the EZ1 DNA Investigator Kit. Concordant profiles were obtained for all samples. Our protocol includes simple punch, wash, and PCR steps, reducing cost and hands-on time in the laboratory. Furthermore, it facilitates...... automation of DNA sequencing....

  12. Sensitive diagnosis of cutaneous leishmaniasis by lesion swab sampling coupled to qPCR

    NARCIS (Netherlands)

    Adams, Emily R.; Gomez, Maria Adelaida; Scheske, Laura; Rios, Ruby; Marquez, Ricardo; Cossio, Alexandra; Albertini, Audrey; Schallig, Henk; Saravia, Nancy Gore

    2014-01-01

    Variation in clinical accuracy of molecular diagnostic methods for cutaneous leishmaniasis (CL) is commonly observed depending on the sample source, the method of DNA recovery and the molecular test. Few attempts have been made to compare these variables. Two swab and aspirate samples from lesions

  13. Comparison of urine samples and penile swabs for detection of human papillomavirus in HIV-negative Dutch men.

    Science.gov (United States)

    Koene, Fleur; Wolffs, Petra; Brink, Antoinette; Dukers-Muijrers, Nicole; Quint, Wim; Bruggeman, Cathrien; Hoebe, Christian

    2016-09-01

    Penile swab sampling is the method of choice when testing for human papillomavirus (HPV) in men. Urine sampling is already used in routine sexually transmitted infections (STI) diagnostics and could provide a less invasive sampling method in men to detect HPV. Therefore we compared detection of HPV types in urine samples and penile swabs by the highly sensitive SPF10-LiPA25 system. First void urine and self-obtained penile swab samples were collected from 120 men, with a mean age of 29.4 years, visiting a STI clinic in South Limburg, the Netherlands. In total 111 of 120 men were included in the analysis. Broad-spectrum HPV DNA amplification and mucosal HPV genotyping were performed using the SPF10 DEIA-LiPA25 system (SPF10 HPV LiPA, V.1). In total 75 (68%) men were positive for HPV in the combined analysis. Sixty-six (59%) paired samples were concordant in being positive or negative. In 39% of the men HPV DNA was detected only in the penile swab. In 2% of the men HPV DNA was detected only in the urine sample. Considering penile swabs as the gold standard, a sensitivity of 41% (95% CI 30% to 53%) and a specificity of 95% (95% CI 81% to 99%) was found. In 6 (5%) urines high risk types were repeatedly found that were not detected in the matching swab. Urine samples are not comparable to penile swabs in the detection of HPV in men. However, the addition of urine samples to penile swabs could be of use in epidemiological or clearance studies. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  14. Primary health clinic toilet/bathroom surface swab sampling can indicate community profile of sexually transmitted infections

    Science.gov (United States)

    Su, Jiunn-Yih; Andersson, Patiyan; Holt, Deborah C.

    2017-01-01

    relative prevalence of gonorrhoeae in central Australia than in northern Australia. Similarly, the regional clinics yielded p values from 0.0088–0.0022. In contrast, swab and notifications data from the sexual health clinic were not correlated. Discussion Strong correlations between swab and notifications were observed. However, there was evidence for limitations of this approach. Despite the correlation observed with the regional clinics data, one clinic yielded zero positive swabs for C. trachomatis, although this STI constituted 25.1% of the corresponding notifications. This could be ascribed to stochastic effects. The lack of correlation observed for sexual health clinic data was also likely due to stochastic effects. It was concluded that toilet/bathroom surface swab sampling has considerable potential for public health surveillance. The approach may be applicable in situations other than primary health clinics, and for targets other than STIs. PMID:28652937

  15. Primary health clinic toilet/bathroom surface swab sampling can indicate community profile of sexually transmitted infections.

    Science.gov (United States)

    Giffard, Philip M; Su, Jiunn-Yih; Andersson, Patiyan; Holt, Deborah C

    2017-01-01

    in central Australia than in northern Australia. Similarly, the regional clinics yielded p values from 0.0088-0.0022. In contrast, swab and notifications data from the sexual health clinic were not correlated. Strong correlations between swab and notifications were observed. However, there was evidence for limitations of this approach. Despite the correlation observed with the regional clinics data, one clinic yielded zero positive swabs for C. trachomatis, although this STI constituted 25.1% of the corresponding notifications. This could be ascribed to stochastic effects. The lack of correlation observed for sexual health clinic data was also likely due to stochastic effects. It was concluded that toilet/bathroom surface swab sampling has considerable potential for public health surveillance. The approach may be applicable in situations other than primary health clinics, and for targets other than STIs.

  16. Diagnosis of Trichomonas vaginalis infection by PCR using vaginal swab samples.

    Science.gov (United States)

    Madico, G; Quinn, T C; Rompalo, A; McKee, K T; Gaydos, C A

    1998-11-01

    Trichomonas vaginalis infection is the most prevalent nonviral sexually transmitted disease (STD) in the world. A PCR test using vaginal swab samples for the detection of T. vaginalis was developed to add T. vaginalis infection to the growing list of STDs that can be detected by DNA amplification techniques. A primer set, BTUB 9/2, was designed to target a well-conserved region in the beta-tubulin genes of T. vaginalis. All strains (15 of 15) of T. vaginalis tested were successfully detected by PCR giving a single predicted product of 112 bp in gel electrophoresis. No such targeted product was amplified with DNA from Trichomonas tenax, Trichomonas gallinae, Chlamydia trachomatis, Neisseria gonorrhoeae, Giardia lamblia, Chilomastix sulcatus, Dientamoeba fragilis, and Entamoeba histolytica. An optimal analytical sensitivity of one T. vaginalis organism per PCR was achieved. Culture, performed with the Inpouch TV culture system, was examined daily with a light microscope to identify T. vaginalis. Twenty-three of 350 (6.6%) vaginal swab samples from women attending an army medical clinic were culture positive for T. vaginalis. Of these culture positive specimens, PCR detected 22 of 23 (96%) with primer set BTUB 9/2, and wet preparation detected only 12 of 23 (52%). Seventeen specimens were BTUB 9/2-PCR positive and culture negative. Ten of these discordant specimens were determined to be as true positive by PCR using primer sets TVA 5-1/6 and/or AP65 A/B, which target different regions in the T. vaginalis genome, and seven were determined to be false positive. The sensitivity of BTUB 9/2-PCR was 97% and the specificity was 98%. The sensitivities of culture and wet preparation were 70 and 36%, respectively. The diagnosis of T. vaginalis infection by PCR is a sensitive and specific method that could be incorporated into a joint strategy for the screening of multiple STDs by using molecular amplification methods.

  17. Evaluation of Rectoanal Mucosal Swab Sampling for Molecular Detection of Enterohemorrhagic Escherichia coli in Beef Cattle.

    Science.gov (United States)

    Agga, Getahun E; Arthur, Terrance M; Hinkley, Susanne; Bosilevac, Joseph M

    2017-04-01

    Cattle are a primary reservoir of enterohemorrhagic Escherichia coli (EHEC), and contaminated beef products are a source of human infections. The U.S. Department of Agriculture Food Safety and Inspection Service declared seven EHEC serogroups (O26, O45, O103, O111, O121, O145, and O157) as adulterants in raw ground beef. Sampling a large number of animals for EHEC surveillance or evaluations of EHEC-focused preharvest interventions requires a convenient and robust sampling method. We evaluated the diagnostic performance of rectoanal mucosal swab (RAMS) for the detection of the top seven EHEC serogroups. Paired fecal grab (FG) and RAMS samples were collected from 176 beef cattle and tested using the NeoSEEK Shiga toxin-producing E. coli (STEC) confirmation method. The prevalence of virulence-associated genes (stx 1 , stx 2 , stx 2c , eae, and nleB) was higher in RAMS than in FG samples. The results of the two methods had poor agreement, as indicated by kappa statistics, for the detection of the seven serogroups. When FG and RAMS results were combined for comparison, RAMS was more sensitive than FG for the detection of serogroups O103 (82% versus 39%), O157 (75% versus 67%), and O45 (79% versus 73%) with similar sensitivity for the detection of serogroup O145 (67%). Serogroups O111 and O121 were detected from one and two samples, respectively, by FG and were not detected by RAMS. Serogroup O26 was not detected with either method. RAMS appears to be equivalent or superior to FG sampling for detection of the top seven EHEC serogroups in the feces of beef cattle with the NeoSEEK STEC confirmation test.

  18. Comparison of culture and a multiplex probe PCR for identifying Mycoplasma species in bovine milk, semen and swab samples.

    Directory of Open Access Journals (Sweden)

    Alysia M Parker

    Full Text Available Mycoplasma spp. are a major cause of mastitis, arthritis and pneumonia in cattle, and have been associated with reproductive disorders in cows. While culture is the traditional method of identification the use of PCR has become more common. Several investigators have developed PCR protocols to detect M. bovis in milk, yet few studies have evaluated other sample types or other important Mycoplasma species. Therefore the objective of this study was to develop a multiplex PCR assay to detect M. bovis, M. californicum and M. bovigenitalium, and evaluate its analytical performance against traditional culture of bovine milk, semen and swab samples. The PCR specificity was determined and the limit of detection evaluated in spiked milk, semen and swabs. The PCR was then compared to culture on 474 field samples from individual milk, bulk tank milk (BTM, semen and swab (vaginal, preputial, nose and eye samples. Specificity analysis produced appropriate amplification for all M. bovis, M. californicum and M. bovigenitalium isolates. Amplification was not seen for any of the other Mollicutes or eubacterial isolates. The limit of detection of the PCR was best in milk, followed by semen and swabs. When all three Mycoplasma species were present in a sample, the limit of detection increased. When comparing culture and PCR, overall there was no significant difference in the proportion of culture and PCR positive samples. Culture could detect significantly more positive swab samples. No significant differences were identified for semen, individual milk or BTM samples. PCR identified five samples with two species present. Culture followed by 16S-23S rRNA sequencing did not enable identification of more than one species. Therefore, the superior method for identification of M. bovis, M. californicum and M. bovigenitalium may be dependent on the sample type being analysed, and whether the identification of multiple target species is required.

  19. Direct PCR amplification of DNA from human bloodstains, saliva, and touch samples collected with microFLOQ®swabs.

    Science.gov (United States)

    Ambers, Angie; Wiley, Rachel; Novroski, Nicole; Budowle, Bruce

    2018-01-01

    Previous studies have shown that nylon flocked swabs outperform traditional fiber swabs in DNA recovery due to their innovative design and lack of internal absorbent core to entrap cellular materials. The microFLOQ ® Direct swab, a miniaturized version of the 4N6 FLOQSwab ® , has a small swab head that is treated with a lysing agent which allows for direct amplification and DNA profiling from sample collection to final result in less than two hours. Additionally, the microFLOQ ® system subsamples only a minute portion of a stain and preserves the vast majority of the sample for subsequent testing or re-analysis, if desired. The efficacy of direct amplification of DNA from dilute bloodstains, saliva stains, and touch samples was evaluated using microFLOQ ® Direct swabs and the GlobalFiler™ Express system. Comparisons were made to traditional methods to assess the robustness of this alternate workflow. Controlled studies with 1:19 and 1:99 dilutions of bloodstains and saliva stains consistently yielded higher STR peak heights than standard methods with 1ng input DNA from the same samples. Touch samples from common items yielded single source and mixed profiles that were consistent with primary users of the objects. With this novel methodology/workflow, no sample loss occurs and therefore more template DNA is available during amplification. This approach may have important implications for analysis of low quantity and/or degraded samples that plague forensic casework. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Detection of Campylobacter jejuni in rectal swab samples from Rousettus amplexicaudatus in the Philippines.

    Science.gov (United States)

    Hatta, Yuki; Omatsu, Tsutomu; Tsuchiaka, Shinobu; Katayama, Yukie; Taniguchi, Satoshi; Masangkay, Joseph S; Puentespina, Roberto; Eres, Eduardo; Cosico, Edison; Une, Yumi; Yoshikawa, Yasuhiro; Maeda, Ken; Kyuwa, Shigeru; Mizutani, Tetsuya

    2016-09-01

    Bats are the second diversity species of mammals and widely distributed in the world. They are thought to be reservoir and vectors of zoonotic pathogens. However, there is scarce report of the evidence of pathogenic bacteria kept in bats. The precise knowledge of the pathogenic bacteria in bat microbiota is important for zoonosis control. Thus, metagenomic analysis targeting the V3-V4 region of the 16S rRNA of the rectal microbiota in Rousettus amplexicaudatus was performed using high throughput sequencing. The results revealed that 103 genera of bacteria including Camplyobacter were detected. Campylobacter was second predominant genus, and Campylobacter coli and Campylobacter jejuni were identified in microbiome of R. amplexicaudatus. Campylobacteriosis is one of the serious bacterial diarrhea in human, and the most often implicated species as the causative agent of campylobacteriosis is C. jejuni. Therefore, we investigated the prevalence of C. jejuni in 91 wild bats with PCR. As a result of PCR assay targeted on 16S-23S intergenic spacer, partial genome of C. jejuni was detected only in five R. amplexicaudatus. This is the first report that C. jejuni was detected in bat rectal swab samples. C. jejuni is the most common cause of campylobacteriosis in humans, transmitted through water and contact with livestock animals. This result indicated that R. amplexicaudatus may be a carrier of C. jejuni.

  1. Primary health clinic toilet/bathroom surface swab sampling can indicate community profile of sexually transmitted infections

    Directory of Open Access Journals (Sweden)

    Philip M. Giffard

    2017-06-01

    known higher relative prevalence of gonorrhoeae in central Australia than in northern Australia. Similarly, the regional clinics yielded p values from 0.0088–0.0022. In contrast, swab and notifications data from the sexual health clinic were not correlated. Discussion Strong correlations between swab and notifications were observed. However, there was evidence for limitations of this approach. Despite the correlation observed with the regional clinics data, one clinic yielded zero positive swabs for C. trachomatis, although this STI constituted 25.1% of the corresponding notifications. This could be ascribed to stochastic effects. The lack of correlation observed for sexual health clinic data was also likely due to stochastic effects. It was concluded that toilet/bathroom surface swab sampling has considerable potential for public health surveillance. The approach may be applicable in situations other than primary health clinics, and for targets other than STIs.

  2. FilmArray Respiratory Panel Assay: Comparison of Nasopharyngeal Swabs and Bronchoalveolar Lavage Samples.

    Science.gov (United States)

    Azadeh, Natalya; Sakata, Kenneth K; Brighton, Anjuli M; Vikram, Holenarasipur R; Grys, Thomas E

    2015-12-01

    The FilmArray respiratory panel (FARP) reliably and rapidly identifies 17 viruses and 3 bacterial pathogens. A nasopharyngeal swab FARP (NP FARP) is performed for many patients with respiratory symptoms. For patients who are acutely ill or immunocompromised or fail to improve, a bronchoalveolar lavage sample FARP (BAL FARP) is performed in addition to the NP FARP. To date, no studies have compared the yield of a BAL FARP with that of an NP FARP. We retrospectively studied all patients who had a BAL FARP within 7 days after an NP FARP between June 2013 and May 2014. Demographic information, comorbidities, FARP results, and all microbiologic data from BAL fluid were collected. Eighty-six patients had a BAL FARP performed within 7 days (mean, 1.6; median, 1) after an NP FARP. Of these, 66 (77%) had concordant BAL and NP FARP results: 15 (23%) had the same pathogen identified from the NP and BAL FARPs, and 51 (77%) had concordant negative FARP results. In 18 of the 86 patients (21%), a pathogen was detected from the NP FARP; of these, 15 (83%) had a concordant match on a subsequent BAL FARP, and the remaining 3 had negative BAL FARPs. In 17 of the 86 patients (20%), pathogens were identified from the BAL FARPs that were not detected by the NP FARPs; of these, 16 (94%) had initial negative NP FARPs. The data suggest that once a pathogen is identified by an NP FARP, a subsequent BAL FARP is unlikely to add new microbiologic information. However, a BAL FARP may provide new, useful microbiologic information when performed within 7 days after a negative NP FARP. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. A simple and novel method for retrieval of Pasteurellaceae from swab samples collected in the field

    DEFF Research Database (Denmark)

    Hansen, Mie Johanne; Bertelsen, Mads Frost; Dietz, Rune

    2013-01-01

    and stored at -20°C. As a control study, 15 samples were collected from the oral cavity of a captive brown bear. One was immediately plated, while the remaining 12 swabs were stored at -20°C for 7 days and multiples of 30 days up to 330 days prior to plating. Two samples were stored without the medium for 7...

  4. Efficiency of noninvasive sampling methods (swab together with Polymerase Chain Reaction (PCR for diagnosing American Tegumentary Leishmaniasis

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    Sara Macente Boni

    Full Text Available ABSTRACT Traditional diagnostic methods used to detect American Tegumentary Leishmaniasis, such as histopathology using biopsy samples, culture techniques, and direct search for parasites, have low sensitivity and require invasive collection procedures. This study evaluates the efficiency of noninvasive sampling methods (swab along with Polymerase Chain Reaction (PCR for diagnosing American Tegumentary Leishmaniasis using skin and mucous samples from 25 patients who had tested positive for leishmaniasis. The outcome of the tests performance on swab samples was compatible with PCR results on biopsy samples. The findings have also shown that PCR-kDNA test is more efficient than PCR-HSP70 and qPCR tests (sensitivity of 92.3%, 40.7%, and 41%, respectively. Given the high sensitivity of the tests and the fact that the sampling method using swabs affords greater patient comfort and safety, it could be said that this method is a promising alternative to conventional biopsy-based methods for the molecular diagnosis of leishmaniasis.

  5. Randomized Comparison of Two Vaginal Self-Sampling Methods for Human Papillomavirus Detection: Dry Swab versus FTA Cartridge.

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    Rosa Catarino

    Full Text Available Human papillomavirus (HPV self-sampling (self-HPV is valuable in cervical cancer screening. HPV testing is usually performed on physician-collected cervical smears stored in liquid-based medium. Dry filters and swabs are an alternative. We evaluated the adequacy of self-HPV using two dry storage and transport devices, the FTA cartridge and swab.A total of 130 women performed two consecutive self-HPV samples. Randomization determined which of the two tests was performed first: self-HPV using dry swabs (s-DRY or vaginal specimen collection using a cytobrush applied to an FTA cartridge (s-FTA. After self-HPV, a physician collected a cervical sample using liquid-based medium (Dr-WET. HPV types were identified by real-time PCR. Agreement between collection methods was measured using the kappa statistic.HPV prevalence for high-risk types was 62.3% (95%CI: 53.7-70.2 detected by s-DRY, 56.2% (95%CI: 47.6-64.4 by Dr-WET, and 54.6% (95%CI: 46.1-62.9 by s-FTA. There was overall agreement of 70.8% between s-FTA and s-DRY samples (kappa = 0.34, and of 82.3% between self-HPV and Dr-WET samples (kappa = 0.56. Detection sensitivities for low-grade squamous intraepithelial lesion or worse (LSIL+ were: 64.0% (95%CI: 44.5-79.8 for s-FTA, 84.6% (95%CI: 66.5-93.9 for s-DRY, and 76.9% (95%CI: 58.0-89.0 for Dr-WET. The preferred self-collection method among patients was s-DRY (40.8% vs. 15.4%. Regarding costs, FTA card was five times more expensive than the swab (~5 US dollars (USD/per card vs. ~1 USD/per swab.Self-HPV using dry swabs is sensitive for detecting LSIL+ and less expensive than s-FTA.International Standard Randomized Controlled Trial Number (ISRCTN: 43310942.

  6. Comparison of human papillomavirus (HPV) detection in urine and cervical swab samples using the HPV GenoArray Diagnostic assay.

    Science.gov (United States)

    Nilyanimit, Pornjarim; Chansaenroj, Jira; Karalak, Anant; Laowahutanont, Piyawat; Junyangdikul, Pairoj; Poovorawan, Yong

    2017-01-01

    Human papillomavirus (HPV) is the leading cause of cervical cancer. Urine-based HPV testing offers a simple and non-invasive method because of its increasing acceptance. A total of 164 pairs of cervical swab and urine samples from Thai women who underwent cervical cancer screening were used for HPV testing with HPV GenoArray Diagnostic Kits. The overall concordance percentage for HPV detection in the cervical swab and urine samples was 65.2%. The HPV genotypes most commonly detected were HPV16 and HPV18. An analysis of the urine samples and a second analysis of the cervical swab samples showed that the differences in the overall HPV detection rate between women with normal and abnormal cytology were not significant (p > 0.05). Urine samples processed with the GenoArray assay is an alternative for women who decline to undergo Pap smear even though it is not ideal as the first-line screening option.

  7. APTIMA assay on SurePath liquid-based cervical samples compared to endocervical swab samples facilitated by a real time database

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    Khader Samer

    2010-01-01

    Full Text Available Background: Liquid-based cytology (LBC cervical samples are increasingly being used to test for pathogens, including: HPV, Chlamydia trachomatis (CT and Neisseria gonorrhoeae (GC using nucleic acid amplification tests. Several reports have shown the accuracy of such testing on ThinPrep (TP LBC samples. Fewer studies have evaluated SurePath (SP LBC samples, which utilize a different specimen preservative. This study was undertaken to assess the performance of the Aptima Combo 2 Assay (AC2 for CT and GC on SP versus endocervical swab samples in our laboratory. Materials and Methods: The live pathology database of Montefiore Medical Center was searched for patients with AC2 endocervical swab specimens and SP Paps taken the same day. SP samples from CT- and/or GC-positive endocervical swab patients and randomly selected negative patients were studied. In each case, 1.5 ml of the residual SP vial sample, which was in SP preservative and stored at room temperature, was transferred within seven days of collection to APTIMA specimen transfer tubes without any sample or patient identifiers. Blind testing with the AC2 assay was performed on the Tigris DTS System (Gen-probe, San Diego, CA. Finalized SP results were compared with the previously reported endocervical swab results for the entire group and separately for patients 25 years and younger and patients over 25 years. Results: SP specimens from 300 patients were tested. This included 181 swab CT-positive, 12 swab GC-positive, 7 CT and GC positive and 100 randomly selected swab CT and GC negative patients. Using the endocervical swab results as the patient′s infection status, AC2 assay of the SP samples showed: CT sensitivity 89.3%, CT specificity 100.0%; GC sensitivity and specificity 100.0%. CT sensitivity for patients 25 years or younger was 93.1%, versus 80.7% for patients over 25 years, a statistically significant difference (P = 0.02. Conclusions: Our results show that AC2 assay of 1.5 ml SP

  8. Noninvasive buccal swab antigen sample and molecular testing provides extended antigen typing for patients with hemoglobinopathies.

    Science.gov (United States)

    Rampersad, Angeli; Hampton, Kisha; Duncan, Natalie; Roberson, Chris; Slayten, Jayanna; Davisson, Suzanne; Aronowitz, Jessica; Shapiro, Amy

    2014-11-01

    To demonstrate the feasibility of performing a noninvasive, molecular-based red blood cell (RBC) antigen test on infants and very young children with sickle cell disease as part of a statewide newborn screening follow-up program. A prospective pilot project was conducted using a noninvasive buccal swab and test kit to perform DNA-based, extended RBC phenotyping in 92 children participating in a newborn hemoglobinopathy screening follow-up program. Reported data include the extended panel of antigens detected by molecular analysis compared with unaffected population estimates. Molecular-based RBC antigen testing was successful, with extended RBC typing generated for all subjects. Molecular testing detected several rare negative or rare positive phenotypes, demonstrating the utility of obtaining an extended antigen panel. This study demonstrates the feasibility of performing antigen testing on buccal swab specimens from children with sickle cell disease as part of a newborn screening follow-up program with the aim of allowing specific unit matching to prevent alloimmunization with RBC transfusions. The general applicability of testing may be limited by a lack of uniform insurance coverage for buccal swab testing, however. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. [Investigation of Chlamydia trachomatis with Cell Culture, DFA and PCR Methods in the Genital Swab Samples of Symptomatic Patients].

    Science.gov (United States)

    Ozüberk, Osman Özüberk; Gökahmetoğlu, Selma; Ozçelik, Bülent; Ekmekçioğlu, Oğuz

    2013-01-01

    Chlamydia trachomatis infection is considered the most prevalent bacterial sexually transmitted disease worldwide. C.trachomatis causes eye infections such as trachoma and newborn inclusion conjunctivitis, newborn pneumonia, genitourinary system infections and suppurative inguinal lymphadenitis namely lymphogranuloma venerum. The aim of this study was to investigate C.trachomatis by direct fluorescent antibody (DFA), polymerase chain reaction (PCR) and cell culture methods in the clinical samples sent to the microbiology laboratory with the prediagnosis of genital infections. A total of 50 swab samples obtained from adult patients (49 female, 1 male) who were admitted to Erciyes University Hospital, Kayseri, Turkey between February-March 2010, were included in the study. C.trachomatis antigens were investigated by a commercial DFA (PathoDx, Remel, USA) method. McCoy cell cultures prepared in microplate wells were used for the isolation of C.trachomatis. The growth of C.trachomatis in cell cultures was confirmed by DFA and iodine staining methods. C.trachomatis DNA was investigated by commercially available PCR (Chlamydia trachomatis 330/740 IC; Sacace, Italy) method. In our study, 4 (8%) of the 50 swab samples were found positive with DFA, 1 (2%) was positive with cell culture, and 1 (2%) was positive with PCR. The only sample that gave positive results with all of the three methods was an urethral swab. Three cervical swab samples that were found positive only with DFA method was evaluated as false positivity. When cell culture was considered as the reference method, the sensitivity and specificity of DFA method were estimated as 100% and 94%, respectively, while those rates for PCR were 100% and 100%, respectively. In conclusion, although cell culture is still the gold standard in the diagnosis of C.trachomatis. infections, since it is time consuming and difficult to apply, more rapid and reliable PCR methods may be applied in diagnosis. DFA method which is

  10. Microbial profiling of cpn60 universal target sequences in artificial mixtures of vaginal bacteria sampled by nylon swabs or self-sampling devices under different storage conditions.

    Science.gov (United States)

    Schellenberg, John J; Oh, Angela Yena; Hill, Janet E

    2017-05-01

    The vaginal microbiome is increasingly characterized by deep sequencing of universal genes. However, there are relatively few studies of how different specimen collection and sample storage and processing influence these molecular profiles. Here, we evaluate molecular microbial community profiles of samples collected using the HerSwab™ self-sampling device, compared to nylon swabs and under different storage conditions. In order to minimize technical variation, mixtures of 11 common vaginal bacteria in simulated vaginal fluid medium were sampled and DNA extracts prepared for massively parallel sequencing of the cpn60 universal target (UT). Three artificial mixtures imitating commonly observed vaginal microbiome profiles were easily distinguished and proportion of sequence reads correlated with the estimated proportion of the organism added to the artificial mixtures. Our results indicate that cpn60 UT amplicon sequencing quantifies the proportional abundance of member organisms in these artificial communities regardless of swab type or storage conditions, although some significant differences were observed between samples that were stored frozen and thawed prior to DNA extraction, compared to extractions from samples stored at room temperature for up to 7days. Our results indicate that an on-the-market device developed for infectious disease diagnostics may be appropriate for vaginal microbiome profiling, an approach that is increasingly facilitated by rapidly dropping deep sequencing costs. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. The comparative utility of oral swabs and probang samples for detection of foot-and-mouth disease virus infection in cattle and pigs.

    Science.gov (United States)

    Stenfeldt, Carolina; Lohse, Louise; Belsham, Graham J

    2013-03-23

    Foot-and-mouth disease virus (FMDV) RNA was measured using quantitative reverse transcription-PCR (qRT-PCR) assays in oral swab and probang samples collected from cattle and pigs during experimental infections with serotype O FMDV. During acute infection, FMDV RNA was measurable in oral swabs as well as in probang samples from both species. FMDV RNA could be detected in oral swabs and probang samples from a time point corresponding to the onset of viremia in directly inoculated animals, whereas animals which were infected through contact exposure had low levels of FMDV RNA in oral swabs before viral RNA could be measured in serum. Analysis of samples collected from cattle persistently infected with FMDV showed that it was not possible to detect FMDV RNA in oral swabs harvested beyond 10 days post infection (dpi), despite the presence of FMDV RNA in probang samples that had been collected as late as 35 dpi. An interesting feature of the persistent infection in the cattle was the apparent decline in the level of FMDV RNA in probang samples after the acute phase of infection, which was followed by a marked rise again (in all the carrier animals) by 28 dpi. Results from this study indicate that qRT-PCR analysis of oral swabs is a useful approach in order to achieve a time efficient and reliable initial diagnosis of acute FMD in cattle and pigs, whereas probang sampling is essential for the detection of cattle that are persistently infected "carriers" of FMDV. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. PCR for detection of Chlamydia trachomatis in endocervical, urethral, rectal, and pharyngeal swab samples obtained from patients attending an STD clinic.

    Science.gov (United States)

    Ostergaard, L; Agner, T; Krarup, E; Johansen, U B; Weismann, K; Gutschik, E

    1997-01-01

    OBJECTIVE: To investigate, by use of the Amplicor PCR in a routine setting, the recovery rate of Chlamydia trachomatis in ano-rectal and pharyngeal swab samples obtained from males and females attending an STD clinic in relation to sexual practices, symptoms, and signs. DESIGN: Data regarding sexual practices, and symptoms and signs related to the rectum and pharynx, were obtained from 196 females and 208 males, including 31 homosexuals and eight bisexuals. Swab samples were obtained from the urethra, rectum, and pharynx from all the patients. An additional endocervical swab sample was obtained from the females. METHODS: All samples were analysed by the Amplicor PCR (Roche). SETTING: Rudolph Bergh's Hospital, a clinic for sexually transmitted diseases situated in the centre of Copenhagen, Denmark. RESULTS: The overall prevalence of urogenital C trachomatis infection was 9.2% (37/404). The specificity of the Amplicor PCR was 100% for both ano-rectal and pharyngeal swab samples. In females three (13%) of the 23 infections were detected only by testing an ano-rectal or throat swab sample. In homosexual males two (67%) of three infections were detected only by the anorectal swab sample. Ano-rectal intercourse without use of condom was reported by 44% of females and by 52% of homosexual males. Fellatio without condom use was reported by 91% of females, and 80% of heterosexual males practised cunnilingus. Pharyngeal infection, however, occurred only in females, and the presence of pharyngeal symptoms or signs seemed predictive for pharyngeal C trachomatis infection, for which the time of incubation or colonisation exceeded 3 months. The presence of ano-rectal signs or symptoms was not predictive for an ano-rectal C trachomatis infection. CONCLUSION: The Amplicor PCR can be used on ano-rectal and pharyngeal swab samples. Ano-rectal swab samples should be obtained in females and homosexual males at high risk of being infected. Pharyngeal samples should be taken in females

  13. A comparison of tonsillar surface swabbing, fine-needle aspiration core sampling, and dissected tonsillar core biopsy culture in children with recurrent tonsillitis.

    Science.gov (United States)

    Sarkar, Saurav; Sil, Abheek; Sarkar, Soma; Sikder, Biswajit

    2017-06-01

    In recurrent tonsillitis, the pathogenic bacteria are harbored in the tonsil core, and therefore cultures of superficial swab samples are not particularly accurate in identifying specific types of core bacteria. On the other hand, the results of fine-needle aspiration (FNA) cultures of core samples have been closely correlated with the findings of core cultures in excised tonsils, and both methods are far superior to surface swabbing. We conducted a prospective study to compare the accuracy of culture findings from tonsillar tissue obtained by surface swabbing, FNA sampling of the tonsil core in situ, and core sampling of the excised tonsil in children with recurrent tonsillitis. Our patient population was made up of 54 children-22 boys and 32 girls, aged 4 to 14 years (mean: 10.7)-who were undergoing elective tonsillectomy during a 1-year period. On the day of surgery, a surface swab, core FNA sample, and dissected core sample were obtained from each patient and sent for culture. Culture showed that the three methods were in agreement in 34 cases (63.0%). In 9 cases (16.7%) the surface swab culture grew different pathogens from those of the two core cultures, and in 3 other cases (5.6%) the surface swab culture was negative while the two core cultures were positive for the same pathogens. In all, the results of core FNA culture and dissected core culture were in agreement in 46 cases (85.2%); in only 4 cases (7.4%) did the core FNA culture fail to accurately identify the causative pathogens. Overall, the sensitivity and specificity of core FNA sampling were 100 and 50% respectively, compared with 82.9 and 30.8% for the superficial tonsillar swab. We conclude that routine culture of surface swab specimens in patients with chronic or recurrent tonsillitis is neither reliable nor valid. We recommend that core FNA sampling be considered the diagnostic method of choice since it can be done on an outpatient basis, it would reliably allow for culture-directed antibiotic

  14. The use of conjunctival swab samples for PCR screening for visceral leishmaniasis in vaccinated dogs O uso de amostras de swab conjuntival para triagem por PCR da leishmaniose visceral em cães vacinados

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    Rodrigo Souza Leite

    2011-03-01

    Full Text Available The polymerase chain reaction (PCR has been shown to provide a rapid and sensitive technique for Leishmania detection. The aim of this study was to evaluate the technique of noninvasive conjunctival swabs (CS as a sampling method for molecular screening for visceral leishmaniasis (VL in a group of 42 police dogs, all of them vaccinated against VL, and to compare the results with those obtained by serological tests. The serological assays were performed independently by three laboratories. Laboratories 1 and 2 were private laboratories and laboratory 3 was the National Reference Laboratory. The first serological screening performed by laboratory 1 showed 15 reactive dogs and 4 indeterminate. Laboratory 2 confirmed only 3 reactive dogs and 2 indeterminate. Laboratory 3 confirmed 7 reactive dogs and 3 indeterminate. The PCR diagnosis using the CS procedure was performed on all 42 animals and was able to detect Leishmania DNA in 17 dogs. The PCR assay confirmed all the cases that were simultaneously reactive in the serological tests by two laboratories. The results showed that the CS technique was a sensitive and practical method for sample collection, thus allowing reliable diagnostic tests through PCR.A PCR (do inglês Polymerase Chain Reaction tem demonstrado ser uma técnica rápida e sensível para detecção de Leishmania. O objetivo deste estudo foi avaliar a técnica não invasiva do swab conjuntival na identificação por PCR de animais infectados em um grupo de 42 cães policiais, todos vacinados contra a Leishmaniose Visceral (VL, e comparar os resultados com aqueles obtidos pelos testes sorológicos. Os ensaios sorológicos foram realizados independentemente por três laboratórios. Os laboratórios 1 e 2 eram privados. O laboratório 3 era o Laboratório de Referência Nacional. A primeira triagem sorológica realizada pelo laboratório 1 apresentou 15 cães reativos e 4 indeterminados. O laboratório 2 confirmou apenas 3 cães reativos

  15. Comparison of plasmid and chromosomal omp1 gene-based PCR and two DNA extraction methods for diagnosing Chlamydia trachomatis in endocervical swab samples

    Directory of Open Access Journals (Sweden)

    Behrouz Taheri Beni

    2012-10-01

    Full Text Available Objective: To evaluate the effectiveness of boiling and proteolytic DNA extraction methods and also to compare the sensitivity of plasmid polymerase chain reaction (PCR and chromosomal omp1 gene PCR for genital Chlamydia trachomatis swab samples in women. Methods: 710 cervical swab samples were obtained from women with symptomatic genital infection at 11 gynecology and obstetric clinics located in Ahvaz, Iran. DNA extraction was performed using proteolysis and boiling manners for all samples. Plasmid PCR and chromosomal omp1 gene primary- and seminested-PCR were then performed separately on extracted DNA in boiling and proteolytic methods. Results: The prevalence of this infection was 17.6% as determined by plasmid-PCR, 13.2% by omp1-primary PCR and 15.8% by omp1-nested PCR. Sensitivities of boiling and proteolytic extraction-directed PCR were 93.6%, and 68.8%, respectively, which are significantly different (P=0.02. The sign of swab-induced bleeding was significantly found to be the most frequent among women infected with this bacterium (P=0.001 and had a sensitivity of 33.6% and a specificity of 80.5%. Conclusions: In order to obtain confident statistical results about sensitivity of each manner, in present study these evaluations were carried out for high numbers of samples (710 samples; high number of samples is statistical advantage of this study in comparing with other studies which were performed with low numbers of samples. Using boilingDNA extraction manner and targeting plasmid sequence for PCR can increase the sensitivity of C. trachomatis diagnosis.

  16. Antibacterial activity of marine culturable bacteria collected from a global sampling of ocean surface waters and surface swabs of marine organisms

    DEFF Research Database (Denmark)

    Gram, Lone; Melchiorsen, Jette; Bruhn, Jesper Bartholin

    2010-01-01

    The purpose of the present study was to isolate marine culturable bacteria with antibacterial activity and hence a potential biotechnological use. Seawater samples (244) and 309 swab samples from biotic or abiotic surfaces were collected on a global Danish marine research expedition (Galathea 3......). Total cell counts at the seawater surface were 5 × 105 to 106 cells/ml, of which 0.1–0.2% were culturable on dilute marine agar (20°C). Three percent of the colonies cultured from seawater inhibited Vibrio anguillarum, whereas a significantly higher proportion (13%) of colonies from inert or biotic...

  17. Efficacy of a Sonicating Swab for Removal and Capture of Listeria monocytogenes in Biofilms on Stainless Steel.

    Science.gov (United States)

    Branck, Tobyn A; Hurley, Matthew J; Prata, Gianna N; Crivello, Christina A; Marek, Patrick J

    2017-06-01

    Listeria monocytogenes is of great concern in food processing facilities because it persists in biofilms, facilitating biotransfer. Stainless steel is commonly used for food contact surfaces and transport containers. L. monocytogenes biofilms on stainless steel served as a model system for surface sampling, to test the performance of a sonicating swab in comparison with a standard cotton swab. Swab performance and consistency were determined using total viable counts. Stainless steel coupons sampled with both types of swabs were examined using scanning electron microscopy, to visualize biofilms and surface structures (i.e., polishing grooves and scratches). Laser scanning confocal microscopy was used to image and to quantitate the biofilms remaining after sampling with each swab type. The total viable counts were significantly higher ( P ≤ 0.05) with the sonicating swab than with the standard swab in each trial. The sonicating swab was more consistent in cell recovery than was the standard swab, with coefficients of variation ranging from 8.9% to 12.3% and from 7.1% to 37.6%, respectively. Scanning electron microscopic imaging showed that biofilms remained in the polished grooves of the coupons sampled with the standard swab but were noticeably absent with the sonicating swab. Percent area measurements of biofilms remaining on the stainless steel coupons showed significantly ( P ≤ 0.05) less biofilm remaining when the sonicating swab was used (median, 1.1%), compared with the standard swab (median, 70.4%). The sonicating swab provided greater recovery of cells, with more consistency, than did the standard swab, and it is employs sonication, suction, and scrubbing. IMPORTANCE Inadequate surface sampling can result in foodborne illness outbreaks from biotransfer, since verification of sanitization protocols relies on surface sampling and recovery of microorganisms for detection and enumeration. Swabbing is a standard method for microbiological sampling of

  18. Canine Skin and Conjunctival Swab Samples for the Detection and Quantification of Leishmania infantum DNA in an Endemic Urban Area in Brazil

    Science.gov (United States)

    de Almeida Ferreira, Sidney; Leite, Rodrigo Souza; Ituassu, Leonardo Trindade; Almeida, Gregório Guilherme; Souza, Daniel Menezes; Fujiwara, Ricardo Toshio; de Andrade, Antero Silva Ribeiro; Melo, Maria Norma

    2012-01-01

    Background We evaluated kDNA PCR/hybridization and quantitative real-time PCR (qPCR) targeting the gene of DNA polymerase of Leishmania infantum for CVL diagnosis and assessment of parasite load in clinical samples obtained invasively and non-invasively. Methodology/Principal Findings Eighty naturally infected dogs from an endemic urban area in Brazil were used. Animals were divided into two groups based on the presence or absence of CVL clinical sings. Skin biopsies, bone marrow, blood and conjunctival swabs samples were collected and submitted to L. infantum DNA detection. In addition, anti-Leishmania antibody titers were measured by Immunofluorescence antibody test. The symptomatic dogs had increased titers compared to asymptomatic dogs (P = 0.025). The frequencies of positive results obtained by kDNA PCR/hybridization for asymptomatic and symptomatic dogs, respectively, were as follows: right conjunctiva, 77.5% and 95.0%; left conjunctiva, 75.0% and 87.5%; skin, 45.0% and 75.0%; bone marrow, 50.0% and 77.5%; and blood, 27.5% and 22.5%. In both groups, the parasite load in the skin samples was the highest (P<0.0001). The parasite loads in the conjunctival swab and bone marrow samples were statistically equivalent within each group. The parasite burden in conjunctival swabs was higher in the dogs with clinical signs than in asymptomatic dogs (P = 0.028). This same relationship was also observed in the bone marrow samples (P = 0.002). No differences in amastigotes load in the skin were detected between the groups. Conclusions The conjunctival swab is a suitable clinical sample for qualitative molecular diagnosis of CVL. The highest parasite burdens were detected in skin regardless of the presence of VL-associated clinical signs. The qPCR results emphasized the role of dogs, particularly asymptomatic dogs, as reservoirs for CVL because of the high cutaneous parasite loads. These results may help to explain the maintenance of high transmission rates and

  19. The role of a commercial enzyme immuno assay antigen detection system for diagnosis of C. trachomatis in genital swab samples

    Directory of Open Access Journals (Sweden)

    A Mukherjee

    2011-01-01

    Full Text Available In the present pilot study, endocervical and urethral swabs collected from 100 patients attending sexually transmitted disease (STD clinics and regional centre for STD in two referral hospitals in New Delhi were analyzed by enzyme immune assay (EIA, polymerase chain reaction (PCR and direct fluorescent antibody (DFA for detection of C. trachomatis. It was found that EIA could detect a very low number of cases (3/100 as against DFA (11/100 and PCR (9/100. Thus, in spite of the widespread availability, lower cost and ease of performance of the enzyme-linked-immunosorbent serologic assay, the present study highlights the need to employ sophisticated diagnostic tools like DFA and PCR for detection of Chlamydia trachomatis in STD patients.

  20. Comparison of false negative rates and limits of detection following macrofoam-swab sampling of Bacillus anthracis surrogates via Rapid Viability PCR and plate culture.

    Science.gov (United States)

    Hutchison, Janine R; Piepel, Greg F; Amidan, Brett G; Hess, Becky M; Sydor, Michael A; Deatherage Kaiser, Brooke L

    2018-01-21

    We evaluated the effects of Bacillus anthracis surrogates, low surface concentrations, surface materials, and assay methods on false-negative rate (FNR) and limit of detection (LOD 95 ) for recovering Bacillus spores using a macrofoam-swab sampling procedure. Bacillus anthracis Sterne or Bacillus atrophaeus Nakamura spores were deposited over a range of low target concentrations (2 - 500 coupon -1 ) onto glass, stainless steel, vinyl tile, and plastic. Samples were assayed using a modified Rapid Viability-PCR (mRV-PCR) method and the traditional plate culture method to obtain FNR and LOD 95 results. Mean FNRs tended to be lower for mRV-PCR compared to culturing, and increased as spore concentration decreased for all surface materials. Surface material, but not B. anthracis surrogate, influenced FNRs with the mRV-PCR method. The mRV-PCR LOD 95 was lowest for glass and highest for vinyl tile. LOD 95 values overall were lower for mRV-PCR than for the culture method. This study adds to the limited data on FNR and LOD 95 for mRV-PCR and culturing methods with low concentrations of B. anthracis sampled from various surface materials by the CDC macrofoam-swab method. These are key inputs for planning characterization and clearance studies for low contamination levels of B. anthracis. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  1. The detection and antimicrobial susceptibility profile of Shigella isolates from meat and swab samples at butchers' shops in Gondar town, Northwest Ethiopia.

    Science.gov (United States)

    Garedew, Legesse; Hagos, Zenabu; Zegeye, Bidir; Addis, Zelalem

    2016-01-01

    Food borne pathogens are major causes of deaths, illnesses and billions of dollars of expenses. The burden of food borne illness is worsened by the ever increasing rate of antimicrobial resistance microbes. Shigella, a bacterial pathogen associated with food, is reported to account for higher prevalence rates of food borne illness in different settings. A cross-sectional study was conducted from February 10 to June 30, 2013, at the butcher houses of Gondar town in the Northwest of Ethiopia to assess the prevalence and antimicrobial susceptibility pattern of Shigella. Cattle raw meat and swab samples from selected critical control points, including knives, chopping boards, and the hands and noses of butchers, were collected and analyzed. The identification of Shigella was carried out using colony characteristics, the Gram reaction, and biochemical tests. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disc diffusion method. The overall hygienic status of the butcher shops was also assessed using a checklist. An observational analysis revealed that the sanitary condition of the butcher shops and their premises was poor. Of 306 samples screened, 10.5% were positive for Shigella. Approximately 7.4% of meat samples and 10.2% of swab samples were contaminated with Shigella. Out of the total Shigella isolates, 90.6%, 46.9%, 18.8% and 9.4% were resistant to ampicillin, amoxicillin, ceftriaxone and tetracycline, respectively. A multidrug resistance pattern was recorded in 27.8% of the isolates. In conclusion, the safety of meat sold at Gondar butchers houses was poor. The identified Shigella isolates showed high levels of drug resistance and multidrug resistance patterns for commonly used antimicrobials in veterinary and human medicine. Practicing wise use of antimicrobials and strict sanitary interventions at different critical control points is strongly recommended, in addition to further in-depth studies to prevent unprecedented consequences from

  2. Swabbing for respiratory viral infections in older patients: a comparison of rayon and nylon flocked swabs.

    Science.gov (United States)

    Hernes, S S; Quarsten, H; Hagen, E; Lyngroth, A L; Pripp, A H; Bjorvatn, B; Bakke, P S

    2011-02-01

    The purpose of this study was to compare the sampling efficacy of rayon swabs and nylon flocked swabs, and of oropharyngeal and nasopharyngeal specimens for the detection of respiratory viruses in elderly patients. Samples were obtained from patients 60 years of age or above who were newly admitted to Sorlandet Hospital Arendal, Norway. The patients were interviewed for current symptoms of a respiratory tract infection. Using rayon swabs and nylon flocked swabs, comparable sets of mucosal samples were harvested from the nasopharynx and the oropharynx. The samples were analysed using real-time polymerase chain reaction (PCR) methods. A total of 223 patients (mean age 74.9 years, standard deviation [SD] 9.0 years) were swabbed and a virus was recovered from 11% of the symptomatic patients. Regardless of the sampling site, a calculated 4.8 times higher viral load (95% confidence interval [CI] 1.3-17, p = 0.017) was obtained using the nylon flocked swabs as compared to the rayon swabs. Also, regardless of the type of swab, a calculated 19 times higher viral load was found in the samples from the nasopharynx as compared to the oropharynx (95% CI 5.4-67.4, p Nylon flocked swabs appear to be more efficient than rayon swabs.

  3. Bacteriological Profile and Antimicrobial Susceptibility Patterns of Bacteria Isolated from Pus/Wound Swab Samples from Children Attending a Tertiary Care Hospital in Kathmandu, Nepal

    Directory of Open Access Journals (Sweden)

    Salu Rai

    2017-01-01

    Full Text Available In Nepal, little is known about the microbiological profile of wound infections in children and their antimicrobial susceptibility patterns. Total of 450 pus/wound swab samples collected were cultured using standard microbiological techniques and the colonies grown were identified with the help of biochemical tests. The antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Methicillin-resistant Staphylococcus aureus isolates were detected by using cefoxitin disc and confirmed by determining minimum inhibitory concentrations (MIC of oxacillin. 264 (59% samples were culture positive. The highest incidence of bacterial infections was noted in the age group of less than 1 year (76%. Out of 264 growth positive samples, Gram-positive bacteria were isolated from 162 (61% samples and Gram-negative bacteria were found in 102 (39% samples. Staphylococcus aureus (99% was the predominant Gram-positive bacteria isolated and Pseudomonas aeruginosa (44% was predominant Gram-negative bacteria. About 19% of S. aureus isolates were found to be methicillin-resistant MIC of oxacillin ranging from 4 μg/mL to 128 μg/mL. Among the children of Nepal, those of age less than 1 year were at higher risk of wound infections by bacteria. Staphylococcus aureus followed by Pseudomonas aeruginosa were the most common bacteria causing wound infections in children.

  4. Antibiotic resistance and biofilm production among the strains of Staphylococcus aureus isolated from pus/wound swab samples in a tertiary care hospital in Nepal.

    Science.gov (United States)

    Belbase, Ankit; Pant, Narayan Dutt; Nepal, Krishus; Neupane, Bibhusan; Baidhya, Rikesh; Baidya, Reena; Lekhak, Binod

    2017-03-23

    The increasing drug resistance along with inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of Staphylococcus aureus are present as the serious problems to the successful treatment of the infections caused by S. aureus. So, the main objectives of this study were to determine the antimicrobial susceptibility patterns along with the rates of inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of S. aureus isolated from pus/wound swab samples. A total of 830 non-repeated pus/wound swab samples were processed using standard microbiological techniques. The colonies grown were identified on the basis of colony morphology, Gram's stain and biochemical tests. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Detection of inducible clindamycin resistance was performed by D test, while detection of methicillin resistant S. aureus (MRSA) was performed by determination of minimum inhibitory concentration of oxacillin by agar dilution method. Similarly, detection of biofilm formation was performed by microtiter plate method. Strains showing resistance to three or more than three different classes of antibiotics were considered multidrug resistant. Total 76 samples showed the growth of S. aureus, among which 36 (47.4%) contained MRSA and 17 (22.4%) samples were found to have S. aureus showing inducible clindamycin resistance. Among the S. aureus isolated from outpatients, 41.9% were MRSA. Highest rates of susceptibility of S. aureus were seen toward linezolid (100%) and vancomycin (100%). Similarly, S. aureus isolated from 35 (46.1%) samples were found to be biofilm producers. Higher rate of inducible clindamycin resistance was seen among MRSA in comparison to methicillin susceptible S. aureus (MSSA). Similarly, higher rates of multidrug resistance and methicillin resistance were found among biofilm producing strains in comparison to biofilm non

  5. Optimized methods for total nucleic acid extraction and quantification of the bat white-nose syndrome fungus, Pseudogymnoascus destructans, from swab and environmental samples

    Science.gov (United States)

    Verant, Michelle; Bohuski, Elizabeth A.; Lorch, Jeffrey M.; Blehert, David

    2016-01-01

    The continued spread of white-nose syndrome and its impacts on hibernating bat populations across North America has prompted nationwide surveillance efforts and the need for high-throughput, noninvasive diagnostic tools. Quantitative real-time polymerase chain reaction (qPCR) analysis has been increasingly used for detection of the causative fungus, Pseudogymnoascus destructans, in both bat- and environment-associated samples and provides a tool for quantification of fungal DNA useful for research and monitoring purposes. However, precise quantification of nucleic acid fromP. destructans is dependent on effective and standardized methods for extracting nucleic acid from various relevant sample types. We describe optimized methodologies for extracting fungal nucleic acids from sediment, guano, and swab-based samples using commercial kits together with a combination of chemical, enzymatic, and mechanical modifications. Additionally, we define modifications to a previously published intergenic spacer–based qPCR test for P. destructans to refine quantification capabilities of this assay.

  6. Comparison of the diagnostic performance of bacterial culture of nasopharyngeal swab and bronchoalveolar lavage fluid samples obtained from calves with bovine respiratory disease

    Science.gov (United States)

    Objective: Examine the culture results, gamithromycin susceptibility, predictive values, and agreement of pooled bilateral nasopharyngeal swabs (NPS) and bronchoalveolar lavages (BAL) for identification of Mannheimia haemolytica genotypes, Pasteurella multocida, and Histophilus somni in calves treat...

  7. False Negative Rates of a Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates via Real-Time PCR

    Energy Technology Data Exchange (ETDEWEB)

    Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Piepel, Gregory F. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Amidan, Brett G. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sydor, Michael A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Kaiser, Brooke L.D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-06-01

    Surface sampling for Bacillus anthracis spores has traditionally relied on detection via bacterial cultivation methods. Although effective, this approach does not provide the level of organism specificity that can be gained through molecular techniques. False negative rates (FNR) and limits of detection (LOD) were determined for two B. anthracis surrogates with modified rapid viability-polymerase chain reaction (mRV-PCR) following macrofoam-swab sampling. This study was conducted in parallel with a previously reported study that analyzed spores using a plate-culture method. B. anthracis Sterne (BAS) or B. atrophaeus Nakamura (BG) spores were deposited onto four surface materials (glass, stainless steel, vinyl tile, and plastic) at nine target concentrations (2 to 500 spores/coupon; 0.078 to 19.375 colony-forming units [CFU] per cm2). Mean FNR values for mRV-PCR analysis ranged from 0 to 0.917 for BAS and 0 to 0.875 for BG and increased as spore concentration decreased (over the concentrations investigated) for each surface material. FNRs based on mRV-PCR data were not statistically different for BAS and BG, but were significantly lower for glass than for vinyl tile. FNRs also tended to be lower for the mRV-PCR method compared to the culture method. The mRV-PCR LOD95 was lowest for glass (0.429 CFU/cm2 with BAS and 0.341 CFU/cm2 with BG) and highest for vinyl tile (0.919 CFU/cm2 with BAS and 0.917 CFU/cm2 with BG). These mRV-PCR LOD95 values were lower than the culture values (BAS: 0.678 to 1.023 CFU/cm2 and BG: 0.820 to 1.489 CFU/cm2). The FNR and LOD95 values reported in this work provide guidance for environmental sampling of Bacillus spores at low concentrations.

  8. PCR based diagnostic assay targeting the beta tubulin gene for the detection of Trichomonas vaginalis infection in vaginal swab samples of symptomatic and asymptomatic women in India

    Directory of Open Access Journals (Sweden)

    Surya Prakash Dwivedi

    2012-10-01

    Full Text Available Objective: To develop an in-house PCR based diagnostic assay for identification of strains isolated from symptomatic and asymptomatic subjects of India, targeting the 毬 -tubulin gene using specific primers. Methods: In the present study a primer set is designed to target a well-conserved region in the beta-tubulin gene of Trichomonas vaginalis (T. vaginalis. All strains of T. vaginalis were tested and successfully detected by PCR yielding a single predicted product of 198 bp in gel electrophoresis, while there was negative response with DNA from Giardia lamblia, Toxoplasma gondii, Leishmania donovani and Entamoeba histolytica. The sensitivity and specificity for a single T. vaginalis cell per PCR was achieved. Axenic Culture, performed with long term axenized T. vaginalis culture system, was routinely examined to identify T. vaginalis. Results: The PCR based investigations with 498 vaginal swab samples from women attending OPD clinics of Halberg Hospital Moradabad and Queen Mary ’s Hospital, Lucknow, India and 17 long term axenic cultures maintained at PGIMER, Chandigarh, India using primer set BTUB 1 & BTUB 2 showed sensitivity and specificity response of 98% and 100%, respectively, while wet preparation in clinically isolated samples responded up to 62.5%. The PCR product sequencing result of symptomatic strains (SS1 of T. vaginalis (744 bp long was submitted to NCBI (Accession No: JF513200. It shows maximum identity 98 % with XM_001284521 Trichomonas vaginalis G-3 beta-tubulin (btub putative partial mRNA. Conclusions: The data gathered in the present study entail that the diagnosis of T. vaginalis infection by PCR may be established as a sensitive and specific protocol, to be incorporated into a joint strategy for the screening of multiple STDs by employing molecular amplification technique. The merits and precautions of the protocol have been discussed.

  9. Recovery Efficiency, False Negative Rate, and Limit of Detection Performance of a Validated Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates

    Energy Technology Data Exchange (ETDEWEB)

    Piepel, Gregory F. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Deatherage Kaiser, Brooke L [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Amidan, Brett G. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sydor, Michael A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Barrett, Christopher A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2015-03-31

    The performance of a macrofoam-swab sampling method was evaluated using Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus Nakamura (BG) spores applied at nine low target amounts (2-500 spores) to positive-control plates and test coupons (2 in. × 2 in.) of four surface materials (glass, stainless steel, vinyl tile, and plastic). Test results from cultured samples were used to evaluate the effects of surrogate, surface concentration, and surface material on recovery efficiency (RE), false negative rate (FNR), and limit of detection. For RE, surrogate and surface material had statistically significant effects, but concentration did not. Mean REs were the lowest for vinyl tile (50.8% with BAS, 40.2% with BG) and the highest for glass (92.8% with BAS, 71.4% with BG). FNR values ranged from 0 to 0.833 for BAS and 0 to 0.806 for BG, with values increasing as concentration decreased in the range tested (0.078 to 19.375 CFU/cm2, where CFU denotes ‘colony forming units’). Surface material also had a statistically significant effect. A FNR-concentration curve was fit for each combination of surrogate and surface material. For both surrogates, the FNR curves tended to be the lowest for glass and highest for vinyl title. The FNR curves for BG tended to be higher than for BAS at lower concentrations, especially for glass. Results using a modified Rapid Viability-Polymerase Chain Reaction (mRV-PCR) analysis method were also obtained. The mRV-PCR results and comparisons to the culture results will be discussed in a subsequent report.

  10. Recovery Efficiency, False Negative Rate, and Limit of Detection Performance of a Validated Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates

    Energy Technology Data Exchange (ETDEWEB)

    Piepel, Gregory F. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Kaiser, Brooke L. D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Amidan, Brett G. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sydor, Michael A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Barrett, Christopher A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-06-16

    The performance of a macrofoam-swab sampling method was evaluated using Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus Nakamura (BG) spores applied at nine low target amounts (2-500 spores) to positive-control plates and test coupons (2 in × 2 in) of four surface materials (glass, stainless steel, vinyl tile, and plastic). Test results from cultured samples were used to evaluate the effects of surrogate, surface concentration, and surface material on recovery efficiency (RE), false negative rate (FNR), and limit of detection. For RE, surrogate and surface material had statistically significant effects, but concentration did not. Mean REs were the lowest for vinyl tile (50.8% with BAS, 40.2% with BG) and the highest for glass (92.8% with BAS, 71.4% with BG). FNR values ranged from 0 to 0.833 for BAS and 0 to 0.806 for BG, with values increasing as concentration decreased in the range tested (0.078 to 19.375 CFU/cm2, where CFU denotes ‘colony forming units’). Surface material also had a statistically significant effect. A FNR-concentration curve was fit for each combination of surrogate and surface material. For both surrogates, the FNR curves tended to be the lowest for glass and highest for vinyl title. The FNR curves for BG tended to be higher than for BAS at lower concentrations, especially for glass. Results using a modified Rapid Viability-Polymerase Chain Reaction (mRV-PCR) analysis method were also obtained. The mRV-PCR results and comparisons to the culture results are discussed in a separate report.

  11. False Negative Rates of a Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates via Real-Time PCR

    Energy Technology Data Exchange (ETDEWEB)

    Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Piepel, Gregory F. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Amidan, Brett G. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Sydor, Michael A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Deatherage Kaiser, Brooke L [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2015-05-01

    Surface sampling for Bacillus anthracis spores has traditionally relied on detection via bacterial cultivation methods. Although effective, this approach does not provide the level of organism specificity that can be gained through molecular techniques. False negative rates (FNR) and limits of detection (LOD) were determined for two B. anthracis surrogates with modified rapid viability-polymerase chain reaction (mRV-PCR) following macrofoam-swab sampling. This study was conducted in parallel with a previously reported study that analyzed spores using a plate-culture method. B. anthracis Sterne (BAS) or B. atrophaeus Nakamura (BG) spores were deposited onto four surface materials (glass, stainless steel, vinyl tile, and plastic) at nine target concentrations (2 to 500 spores/coupon; 0.078 to 19.375 colony-forming units [CFU] per cm²). Mean FNR values for mRV-PCR analysis ranged from 0 to 0.917 for BAS and 0 to 0.875 for BG and increased as spore concentration decreased (over the concentrations investigated) for each surface material. FNRs based on mRV-PCR data were not statistically different for BAS and BG, but were significantly lower for glass than for vinyl tile. FNRs also tended to be lower for the mRV-PCR method compared to the culture method. The mRV-PCR LOD₉₅ was lowest for glass (0.429 CFU/cm² with BAS and 0.341 CFU/cm² with BG) and highest for vinyl tile (0.919 CFU/cm² with BAS and 0.917 CFU/cm² with BG). These mRV-PCR LOD₉₅ values were lower than the culture values (BAS: 0.678 to 1.023 CFU/cm² and BG: 0.820 to 1.489 CFU/cm²). The FNR and LOD₉₅ values reported in this work provide guidance for environmental sampling of Bacillus spores at low concentrations.

  12. Field Validation of SYBR Green- and TaqMan-Based Real-Time PCR Using Biopsy and Swab Samples To Diagnose American Tegumentary Leishmaniasis in an Area Where Leishmania (Viannia) braziliensis Is Endemic.

    Science.gov (United States)

    Gomes, Ciro Martins; Cesetti, Mariana Vicente; de Paula, Natália Aparecida; Vernal, Sebastián; Gupta, Gaurav; Sampaio, Raimunda Nonata Ribeiro; Roselino, Ana Maria

    2017-02-01

    The precise diagnosis of American tegumentary leishmaniasis (ATL) is an essential task due to the disease's associated morbidity. A noninvasive, extremely sensitive, and highly specific exam is critical, particularly for mucosal leishmaniasis (ML), in which a low parasite quantity is expected. We aimed to compare the diagnostic accuracy of swab and biopsy sample analysis using SYBR Green- and TaqMan-based real-time PCR (qPCR) assays with that of a composite reference standard consisting of the Montenegro skin test, serology, histopathology, smears, culture, and conventional PCR. In total, 55 patients with ATL (ML, 18 patients; cutaneous leishmaniasis [CL], 37 patients) and 36 patients without ATL were studied. qPCR analysis of swabs was more accurate when using SYBR Green (87.88%; 95% confidence interval [CI], 77.86 to 93.73 patients) than when using TaqMan (78.79%; 95% CI, 67.49 to 86.92%) (P = 0.031). SYBR Green (84.72%; 95% CI, 74.68 to 91.25%) was also more accurate than TaqMan (73.61%; 95% CI, 62.42 to 82.41%) for biopsy samples (P = 0.008). All qPCR methods were 100% specific. Swabs and biopsy specimens had similar sensitivity when using the same chemistry (P = 0.125 for SYBR Green and P = 0.625 for TaqMan). Moreover, qPCR achieved better performance than most existing techniques used for the diagnosis of ATL and also detected the Leishmania parasite in a greater proportion of patients than the associated histopathology, smear, culture, and conventional PCR techniques did. Swabs therefore represent a useful diagnostic tool because they not only are noninvasive but also can achieve an accuracy similar to that of biopsy samples. The high accuracy of SYBR Green-based qPCR may also reduce the requirement for associated parasitological tests for ATL diagnosis. Copyright © 2017 American Society for Microbiology.

  13. Sampling: Making Electronic Discovery More Cost Effective

    Directory of Open Access Journals (Sweden)

    Milton Luoma

    2011-06-01

    Full Text Available With the huge volumes of electronic data subject to discovery in virtually every instance of litigation, time and costs of conducting discovery have become exceedingly important when litigants plan their discovery strategies.  Rather than incurring the costs of having lawyers review every document produced in response to a discovery request in search of relevant evidence, a cost effective strategy for document review planning is to use statistical sampling of the database of documents to determine the likelihood of finding relevant evidence by reviewing additional documents.  This paper reviews and discusses how sampling can be used to make document review more cost effective by considering issues such as an appropriate sample size, how to develop a sampling strategy, and taking into account the potential value of the litigation in relation to the costs of additional discovery efforts. 

  14. On sampling fractions and electron shower shapes

    Energy Technology Data Exchange (ETDEWEB)

    Peryshkin, Alexander; Raja, Rajendran; /Fermilab

    2011-12-01

    We study the usage of various definitions of sampling fractions in understanding electron shower shapes in a sampling multilayer electromagnetic calorimeter. We show that the sampling fractions obtained by the conventional definition (I) of (average observed energy in layer)/(average deposited energy in layer) will not give the best energy resolution for the calorimeter. The reason for this is shown to be the presence of layer by layer correlations in an electromagnetic shower. The best resolution is obtained by minimizing the deviation from the total input energy using a least squares algorithm. The 'sampling fractions' obtained by this method (II) are shown to give the best resolution for overall energy. We further show that the method (II) sampling fractions are obtained by summing the columns of a non-local {lambda} tensor that incorporates the correlations. We establish that the sampling fractions (II) cannot be used to predict the layer by layer energies and that one needs to employ the full {lambda} tensor for this purpose. This effect is again a result of the correlations.

  15. Enumeration of Escherichia coli in swab samples from pre- and post-chilled pork and lamb carcasses using 3M™ Petrifilm™ Select E. coli and Simplate® Coliforms/E. coli.

    Science.gov (United States)

    Hauge, Sigrun J; Østensvik, Øyvin; Monshaugen, Marte; Røtterud, Ole-Johan; Nesbakken, Truls; Alvseike, Ole

    2017-08-01

    The aim of the study was to compare two analytical methods; 3M Petrifilm™ Select E. coli and SimPlate® Coliforms &E. coli, for detection and enumeration of E. coli using swab samples from naturally contaminated pork and lamb carcasses that were collected before and after chilling. Blast chilling was used for pork carcasses. Swab samples (n=180) were collected from 60 warm and 60 chilled pork carcasses, and 30 warm and 30 chilled lamb carcasses, and analysed in parallel. The concordance correlation coefficient between Petrifilm and SimPlate was 0.89 for pork and 0.81 for lamb carcasses. However, the correlation was higher for warm carcasses (0.90) than chilled carcasses (0.72). For chilled lamb carcasses, the correlation was only 0.50, and SimPlate gave slightly higher results than Petrifilm (P=0.09). Slower chilling gave slightly lesser agreement between methods than for blast chilling, however, both Petrifilm and SimPlate methodologies are suitable and recommended for use in small laboratories in abattoirs. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Detection of influenza A virus from live-bird market poultry swab samples in China by a pan-IAV, one-step reverse-transcription FRET-PCR.

    Science.gov (United States)

    Luan, Lu; Sun, Zhihao; Kaltenboeck, Bernhard; Huang, Ke; Li, Min; Peng, Daxin; Xu, Xiulong; Ye, Jianqiang; Li, Jing; Guo, Weina; Wang, Chengming

    2016-07-22

    The persistent public health threat of animal to human transmission of influenza A virus (IAV) has stimulated interest in rapid and accurate detection of all IAV subtypes in clinical specimens of animal origin. In this study, a new set of primers and probes was designed for one-step pan-IAV reverse-transcription fluorescence resonance energy transfer (FRET)-PCR. The detection limit of one-step pan-IAV RT FRET-PCR was 10 copies of the matrix gene per reaction, and proved to be equivalent or superior to virus isolation in detecting nine IAV subtypes. Application of the pan-IAV RT FRET-PCR to oral-pharyngeal and cloacal swab specimens collected from healthy poultry in 34 live bird markets in 24 provinces of China revealed that 9.2% of the animals (169/1,839) or 6.3% of their oral-pharyngeal or cloacal swabs (233/3,678) were positive for IAV, and 56.8% of IAV-positive samples were of the H9N2 subtype. Paralleling detection of IAV in H9N2-infected SPF chickens and chickens from LBM showed that pan-IAV FRET-PCR had a higher detection limit than virus isolation in eggs while the results by FRET-PCR and virus isolation overall matched. It is expected that this strategy can be useful for facile surveillance for IAV in clinical samples from a variety of sources.

  17. Development of an ELISA for evaluation of swab recovery efficiencies of bovine serum albumin.

    Directory of Open Access Journals (Sweden)

    Nadja Sparding

    Full Text Available After a potential biological incident the sampling strategy and sample analysis are crucial for the outcome of the investigation and identification. In this study, we have developed a simple sandwich ELISA based on commercial components to quantify BSA (used as a surrogate for ricin with a detection range of 1.32-80 ng/mL. We used the ELISA to evaluate different protein swabbing procedures (swabbing techniques and after-swabbing treatments for two swab types: a cotton gauze swab and a flocked nylon swab. The optimal swabbing procedure for each swab type was used to obtain recovery efficiencies from different surface materials. The surface recoveries using the optimal swabbing procedure ranged from 0-60% and were significantly higher from nonporous surfaces compared to porous surfaces. In conclusion, this study presents a swabbing procedure evaluation and a simple BSA ELISA based on commercial components, which are easy to perform in a laboratory with basic facilities. The data indicate that different swabbing procedures were optimal for each of the tested swab types, and the particular swab preference depends on the surface material to be swabbed.

  18. Nylon flocked swab severely reduces Hexagon Obti sensibility.

    Science.gov (United States)

    Frippiat, Christophe; De Roy, Gilbert; Fontaine, Louis-Marie; Dognaux, Sophie; Noel, Fabrice; Heudt, Laeticia; Lepot, Laurent

    2015-02-01

    Hexagon Obti immunological blood test and flocked swab are widely used in forensic laboratories. Nevertheless, up to now, no compatibility tests have been published between sampling with the ethylene oxide treated flocked swab and the Hexagon Obti blood detection strip. In this study, we investigated this compatibility. Our work shows that sampling with ethylene oxide treated flocked swab reduces by a factor of at least 100 the detection threshold of blood using the Hexagon Obti immunological test. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  19. EVA-Compatible Microbial Swab Tool

    Science.gov (United States)

    Rucker, Michelle A.

    2016-01-01

    When we send humans to search for life on Mars, we'll need to know what we brought with us versus what may already be there. To ensure our crewed spacecraft meet planetary protection requirements—and to protect our science from human contamination—we'll need to know whether micro-organisms are leaking/venting from our ships and spacesuits. This is easily done by swabbing external vents and suit surfaces for analysis, but requires a specialized tool for the job. Engineers at the National Aeronautics and Space Administration (NASA) recently developed an Extravehicular Activity (EVA)-compatible swab tool that can be used to sample current space suits and life support systems. Data collected now will influence Mars life support and EVA hardware early in the planning process, before design changes become difficult and expensive.NASA’s EVA swab tool pairs a Space Shuttle-era tool handle with a commercially available swab tip mounted into a custom-designed end effector. A glove-compatible release mechanism allows the handle to quickly switch between swab tips, much like a shaving razor handle can snap onto a disposable blade cartridge. Swab tips are stowed inside individual sterile containers, each fitted with a microbial filter that allows the container to equalize atmospheric pressure, but prevents cabin contaminants from rushing into the container when passing from the EVA environment into a pressurized cabin. A bank of containers arrayed inside a tool caddy allows up to six individual samples to be collected during a given spacewalk.NASA plans to use the tool in 2016 to collect samples from various spacesuits during ground testing to determine what (if any) human-borne microbial contamination leaks from the suit under simulated thermal vacuum conditions. Next, the tool will be used on board the International Space Station to assess the types of microbial contaminants found on external environmental control and life support system vents. Data will support

  20. [Optimalisation of the antibiotic policy in The Netherlands. XI. The national electronic antibiotic guide'SWAB-ID' for use in hospitals

    NARCIS (Netherlands)

    Vonderen, M.G. van; Gyssens, I.C.J.; Hartwig, N.G.; Kullberg, B.J.; Leverstein-van Hall, M.A.; Natsch, S.S.; Prins, J.M.

    2006-01-01

    The 'Stichting Werkgroep Antibioticabeleid' (Dutch Working Party on Antibiotic Policy) has developed an electronic national antibiotic guide for the antibiotic treatment and prophylaxis of common infectious diseases in hospitals. This guide also contains information on the most important

  1. Composition quantification of electron-transparent samples by backscattered electron imaging in scanning electron microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Müller, E., E-mail: erich.mueller@kit.edu; Gerthsen, D.

    2017-02-15

    The contrast of backscattered electron (BSE) images in scanning electron microscopy (SEM) depends on material parameters which can be exploited for composition quantification if some information on the material system is available. As an example, the In-concentration in thin In{sub x}Ga{sub 1−x}As layers embedded in a GaAs matrix is analyzed in this work. The spatial resolution of the technique is improved by using thin electron-transparent specimens instead of bulk samples. Although the BSEs are detected in a comparably small angular range by an annular semiconductor detector, the image intensity can be evaluated to determine the composition and local thickness of the specimen. The measured intensities are calibrated within one single image to eliminate the influence of the detection and amplification system. Quantification is performed by comparison of experimental and calculated data. Instead of using time-consuming Monte-Carlo simulations, an analytical model is applied for BSE-intensity calculations which considers single electron scattering and electron diffusion. - Highlights: • Sample thickness and composition are quantified by backscattered electron imaging. • A thin sample is used to achieve spatial resolution of few nanometers. • Calculations are carried out with a time-saving electron diffusion model. • Small differences in atomic number and density detected at low electron energies.

  2. Potential concerns with analytical Methods Used for the detection of Batrachochytrium salamandrivorans from archived DNA of amphibian swab samples, Oregon, USA

    Science.gov (United States)

    Iwanowicz, Deborah; Olson, Deanna H.; Adams, Michael J.; Adams, Cynthia; Anderson, Chauncey; Blaustein, Andrew R; Densmore, Christine L.; Figiel, Chester; Schill, William B.; Chestnut, Tara

    2017-01-01

    Taxonomic identification of pollen has historically been accomplished via light microscopy but requires specialized knowledge and reference collections, particularly when identification to lower taxonomic levels is necessary. Recently, next-generation sequencing technology has been used as a cost-effective alternative for identifying bee-collected pollen; however, this novel approach has not been tested on a spatially or temporally robust number of pollen samples. Here, we compare pollen identification results derived from light microscopy and DNA sequencing techniques with samples collected from honey bee colonies embedded within a gradient of intensive agricultural landscapes in the Northern Great Plains throughout the 2010–2011 growing seasons. We demonstrate that at all taxonomic levels, DNA sequencing was able to discern a greater number of taxa, and was particularly useful for the identification of infrequently detected species. Importantly, substantial phenological overlap did occur for commonly detected taxa using either technique, suggesting that DNA sequencing is an appropriate, and enhancing, substitutive technique for accurately capturing the breadth of bee-collected species of pollen present across agricultural landscapes. We also show that honey bees located in high and low intensity agricultural settings forage on dissimilar plants, though with overlap of the most abundantly collected pollen taxa. We highlight practical applications of utilizing sequencing technology, including addressing ecological issues surrounding land use, climate change, importance of taxa relative to abundance, and evaluating the impact of conservation program habitat enhancement efforts.

  3. Rectal swabs for analysis of the intestinal microbiota.

    Directory of Open Access Journals (Sweden)

    Andries E Budding

    Full Text Available The composition of the gut microbiota is associated with various disease states, most notably inflammatory bowel disease, obesity and malnutrition. This underlines that analysis of intestinal microbiota is potentially an interesting target for clinical diagnostics. Currently, the most commonly used sample types are feces and mucosal biopsy specimens. Because sampling method, storage and processing of samples impact microbiota analysis, each sample type has its own limitations. An ideal sample type for use in routine diagnostics should be easy to obtain in a standardized fashion without perturbation of the microbiota. Rectal swabs may satisfy these criteria, but little is known about microbiota analysis on these sample types. In this study we investigated the characteristics and applicability of rectal swabs for gut microbiota profiling in a clinical routine setting in patients presenting with various gastro-intestinal disorders. We found that rectal swabs appeared to be a convenient means of sampling the human gut microbiota. Swabs can be performed on demand, whenever a patient presents; swab-derived microbiota profiles are reproducible, whether they are gathered at home by patients or by medical professionals in an outpatient setting and may be ideally suited for clinical diagnostics and large-scale studies.

  4. Rectal swabs for analysis of the intestinal microbiota.

    Science.gov (United States)

    Budding, Andries E; Grasman, Matthijs E; Eck, Anat; Bogaards, Johannes A; Vandenbroucke-Grauls, Christina M J E; van Bodegraven, Adriaan A; Savelkoul, Paul H M

    2014-01-01

    The composition of the gut microbiota is associated with various disease states, most notably inflammatory bowel disease, obesity and malnutrition. This underlines that analysis of intestinal microbiota is potentially an interesting target for clinical diagnostics. Currently, the most commonly used sample types are feces and mucosal biopsy specimens. Because sampling method, storage and processing of samples impact microbiota analysis, each sample type has its own limitations. An ideal sample type for use in routine diagnostics should be easy to obtain in a standardized fashion without perturbation of the microbiota. Rectal swabs may satisfy these criteria, but little is known about microbiota analysis on these sample types. In this study we investigated the characteristics and applicability of rectal swabs for gut microbiota profiling in a clinical routine setting in patients presenting with various gastro-intestinal disorders. We found that rectal swabs appeared to be a convenient means of sampling the human gut microbiota. Swabs can be performed on demand, whenever a patient presents; swab-derived microbiota profiles are reproducible, whether they are gathered at home by patients or by medical professionals in an outpatient setting and may be ideally suited for clinical diagnostics and large-scale studies.

  5. EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

    Science.gov (United States)

    Rucker, Michelle A.; Hood, Drew; Walker, Mary; Venkateswaran, Kasthuri J.; Schuerger, Andrew C.

    2018-01-01

    When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements-and to protect our science from human contamination-we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. Engineers at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each attachment. This allowed the tool handle to snap onto a fresh swab attachment much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab attachment is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and repressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between

  6. Detecting and quantifying influenza virus with self- versus investigator-collected mid-turbinate nasal swabs.

    Science.gov (United States)

    Granados, Andrea; Quach, Susan; McGeer, Allison; Gubbay, Jonathan B; Kwong, Jeffrey C

    2017-07-01

    We compared pairs of self- and investigator-collected mid-turbinate nasal swabs to detect and quantify influenza viral loads. We used RNase P, which reflects presence of human cells to determine adequate sample collection. Sixteen pairs of influenza-positive swabs and 25 pairs of influenza-negative swabs were included in this study. The median influenza A viral loads for self- and investigator-collected swabs were 1.68 and 1.67 log10 copies/mL, respectively (P = 0.96). RNase P loads were also similar between self- and investigator-collected swabs (P = 0.51). Self-collected mid-turbinate nasal swabs yield comparable viral loads to investigator-collected swabs, and therefore might be considered for research and clinical management. © 2017 Wiley Periodicals, Inc.

  7. False-negative rate, limit of detection and recovery efficiency performance of a validated macrofoam-swab sampling method for low surface concentrations of Bacillus anthracis Sterne and Bacillus atrophaeus spores

    Energy Technology Data Exchange (ETDEWEB)

    Piepel, G. F. [Applied Statistics and Computational Sciences, Pacific Northwest National Laboratory, Richland WA USA; Deatherage Kaiser, B. L. [Chemical and Biological Signature Science Group, Pacific Northwest National Laboratory, Richland WA USA; Amidan, B. G. [Applied Statistics and Computational Sciences, Pacific Northwest National Laboratory, Richland WA USA; Sydor, M. A. [Chemical and Biological Signature Science Group, Pacific Northwest National Laboratory, Richland WA USA; Barrett, C. A. [Analytical Chemistry of Nuclear Materials, Pacific Northwest National Laboratory, Richland WA USA; Hutchison, J. R. [Chemical and Biological Signature Science Group, Pacific Northwest National Laboratory, Richland WA USA

    2016-05-06

    The performance of a macrofoam-swab sampling method was evaluated using Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus Nakamura (BG) spores applied at nine low target amounts (2-500 spores) to positive-control plates and test coupons (2 in × 2 in) of four surface materials (glass, stainless steel, vinyl tile, and plastic). Test results from cultured samples were used to evaluate the effects of surrogate, surface concentration, and surface material on recovery efficiency (RE), false negative rate (FNR), and limit of detection. For RE, surrogate and surface material had statistically significant effects, but concentration did not. Mean REs were the lowest for vinyl tile (50.8% with BAS and 40.2% with BG) and the highest for glass (92.8% with BAS and 71.4% with BG). FNR values ranged from 0 to 0.833 for BAS and 0 to 0.806 for BG; values increased as concentration decreased in the range tested (0.078 to 19.375 CFU/cm2). Surface material also had a statistically significant effect. A FNR-concentration curve was fit for each combination of surrogate and surface material. For both surrogates, the FNR curves tended to be the lowest for glass and highest for vinyl title. The FNR curves for BG tended to be higher than for BAS at lower concentrations, especially for glass. Results using a modified Rapid Viability-Polymerase Chain Reaction (mRV-PCR) analysis method were also obtained. The mRV-PCR results and comparisons to the culture results will be discussed in a subsequent article.

  8. Transmission electron microscope sample holder with optical features

    Science.gov (United States)

    Milas, Mirko [Port Jefferson, NY; Zhu, Yimei [Stony Brook, NY; Rameau, Jonathan David [Coram, NY

    2012-03-27

    A sample holder for holding a sample to be observed for research purposes, particularly in a transmission electron microscope (TEM), generally includes an external alignment part for directing a light beam in a predetermined beam direction, a sample holder body in optical communication with the external alignment part and a sample support member disposed at a distal end of the sample holder body opposite the external alignment part for holding a sample to be analyzed. The sample holder body defines an internal conduit for the light beam and the sample support member includes a light beam positioner for directing the light beam between the sample holder body and the sample held by the sample support member.

  9. A Comparison of Dacron versus Flocked Nylon Swabs for Anal Cytology Specimen Collection

    Science.gov (United States)

    Gage, Julia C.; Ghosh, Arpita; Borgonovo, Sylvia; Follansbee, Stephen; Wentzensen, Nicolas; Gravitt, Patti E.; Grabe, Niels; Lahrmann, Bernd; Castle, Philip E.

    2011-01-01

    Objectives We compared the performance of commonly used Dacron versus flocked nylon swabs for anal cytology. Study Design From 23 HIV-positive men screened at Kaiser Permanente San Francisco (San Francisco, Calif., USA), 2 anal specimens were collected, 1 with each swab in random order, and placed into liquid cytology medium. Specimens were tested for cellularity by quantifying a genomic DNA (erv-3). The number of cells was assessed from prepared slides by automated image analysis. Performance was compared between swabs using 2-sample t tests and standard crossover trial analysis methods accounting for period effect. Results Flocked swabs collected slightly more erv-3 cells than Dacron for the first sample although not significantly (p = 0.18) and a similar number of erv-3 cells for the second sample (p = 0.85). Flocked swabs collected slightly more cells per slide than the Dacron swabs at both time periods although this was only significant in the second time period (p = 0.42 and 0.03 for first and second periods, respectively). In crossover trial analysis, flocked swabs outperformed Dacron for cell count per slide based on slide imaging (p = 0.03), but Dacron and flocked swabs performed similarly based on erv-3 quantification (p = 0.14). Conclusions Further studies should determine whether flocked swabs increase the representation of diagnostically important cells compared to Dacron. PMID:21791907

  10. Swabs to genomes: a comprehensive workflow

    Directory of Open Access Journals (Sweden)

    Madison I. Dunitz

    2015-05-01

    Full Text Available The sequencing, assembly, and basic analysis of microbial genomes, once a painstaking and expensive undertaking, has become much easier for research labs with access to standard molecular biology and computational tools. However, there are a confusing variety of options available for DNA library preparation and sequencing, and inexperience with bioinformatics can pose a significant barrier to entry for many who may be interested in microbial genomics. The objective of the present study was to design, test, troubleshoot, and publish a simple, comprehensive workflow from the collection of an environmental sample (a swab to a published microbial genome; empowering even a lab or classroom with limited resources and bioinformatics experience to perform it.

  11. Scanning Electron Microscopy with Samples in an Electric Field

    Science.gov (United States)

    Frank, Ludĕk; Hovorka, Miloš; Mikmeková, Šárka; Mikmeková, Eliška; Müllerová, Ilona; Pokorná, Zuzana

    2012-01-01

    The high negative bias of a sample in a scanning electron microscope constitutes the “cathode lens” with a strong electric field just above the sample surface. This mode offers a convenient tool for controlling the landing energy of electrons down to units or even fractions of electronvolts with only slight readjustments of the column. Moreover, the field accelerates and collimates the signal electrons to earthed detectors above and below the sample, thereby assuring high collection efficiency and high amplification of the image signal. One important feature is the ability to acquire the complete emission of the backscattered electrons, including those emitted at high angles with respect to the surface normal. The cathode lens aberrations are proportional to the landing energy of electrons so the spot size becomes nearly constant throughout the full energy scale. At low energies and with their complete angular distribution acquired, the backscattered electron images offer enhanced information about crystalline and electronic structures thanks to contrast mechanisms that are otherwise unavailable. Examples from various areas of materials science are presented.

  12. Sample Preparation for Electron Probe Microanalysis-Pushing the Limits.

    Science.gov (United States)

    Geller, Joseph D; Engle, Paul D

    2002-01-01

    There are two fundamental considerations in preparing samples for electron probe microanalysis (EPMA). The first one may seem obvious, but we often find it is overlooked. That is, the sample analyzed should be representative of the population from which it comes. The second is a direct result of the assumptions in the calculations used to convert x-ray intensity ratios, between the sample and standard, to concentrations. Samples originate from a wide range of sources. During their journey to being excited under the electron beam for the production of x rays there are many possibilities for sample alteration. Handling can contaminate samples by adding extraneous matter. In preparation, the various abrasives used in sizing the sample by sawing, grinding and polishing can embed themselves. The most accurate composition of a contaminated sample is, at best, not representative of the original sample; it is misleading. Our laboratory performs EPMA analysis on customer submitted samples and prepares over 250 different calibration standards including pure elements, compounds, alloys, glasses and minerals. This large variety of samples does not lend itself to mass production techniques, including automatic polishing. Our manual preparation techniques are designed individually for each sample. The use of automated preparation equipment does not lend itself to this environment, and is not included in this manuscript. The final step in quantitative electron probe microanalysis is the conversion of x-ray intensities ratios, known as the "k-ratios," to composition (in mass fraction or atomic percent) and/or film thickness. Of the many assumptions made in the ZAF (where these letters stand for atomic number, absorption and fluorescence) corrections the localized geometry between the sample and electron beam, or takeoff angle, must be accurately known. Small angular errors can lead to significant errors in the final results. The sample preparation technique then becomes very

  13. Sample Preparation for Electron Probe Microanalysis—Pushing the Limits

    Science.gov (United States)

    Geller, Joseph D.; Engle, Paul D.

    2002-01-01

    There are two fundamental considerations in preparing samples for electron probe microanalysis (EPMA). The first one may seem obvious, but we often find it is overlooked. That is, the sample analyzed should be representative of the population from which it comes. The second is a direct result of the assumptions in the calculations used to convert x-ray intensity ratios, between the sample and standard, to concentrations. Samples originate from a wide range of sources. During their journey to being excited under the electron beam for the production of x rays there are many possibilities for sample alteration. Handling can contaminate samples by adding extraneous matter. In preparation, the various abrasives used in sizing the sample by sawing, grinding and polishing can embed themselves. The most accurate composition of a contaminated sample is, at best, not representative of the original sample; it is misleading. Our laboratory performs EPMA analysis on customer submitted samples and prepares over 250 different calibration standards including pure elements, compounds, alloys, glasses and minerals. This large variety of samples does not lend itself to mass production techniques, including automatic polishing. Our manual preparation techniques are designed individually for each sample. The use of automated preparation equipment does not lend itself to this environment, and is not included in this manuscript. The final step in quantitative electron probe microanalysis is the conversion of x-ray intensities ratios, known as the “k-ratios,” to composition (in mass fraction or atomic percent) and/or film thickness. Of the many assumptions made in the ZAF (where these letters stand for atomic number, absorption and fluorescence) corrections the localized geometry between the sample and electron beam, or takeoff angle, must be accurately known. Small angular errors can lead to significant errors in the final results. The sample preparation technique then becomes very

  14. A comparison of DNA collection and retrieval from two swab types (cotton and nylon flocked swab) when processed using three QIAGEN extraction methods.

    Science.gov (United States)

    Brownlow, Robert J; Dagnall, Kathryn E; Ames, Carole E

    2012-05-01

    The Metropolitan Police Service currently uses cotton swabs to retrieve DNA for forensic profiling. Recently, a new nylon flocked swab type has become available from Copan (MicroRheologics, Brescia, Italy) that it is claimed, offers increased sample recovery and release yields. If true, the flocked swab may have important applications in DNA evidence retrieval. This study examines the DNA retrieval capability of cotton and nylon flocked swabs when extracted using three common extraction platforms (QIAcube, BioRobot EZ1 and manually processed QIAamp DNA investigator kit). Results indicate that both swab types are capable of recovering high percentages of DNA (>50%); however, the extraction platform selected was shown to have a significant effect upon DNA retrieval. Across all experiments, the cotton swab combined with the spin-column extractions was shown to be most effective, with the nylon swab and BioRobot EZ1 combination being the least effective. These findings illustrate the importance of extraction method selection. © 2011 American Academy of Forensic Sciences.

  15. Micro Electron MicroProbe and Sample Analyzer

    Science.gov (United States)

    Manohara, Harish; Bearman, Gregory; Douglas, Susanne; Bronikowski, Michael; Urgiles, Eduardo; Kowalczyk, Robert; Bryson, Charles

    2009-01-01

    A proposed, low-power, backpack-sized instrument, denoted the micro electron microprobe and sample analyzer (MEMSA), would serve as a means of rapidly performing high-resolution microscopy and energy-dispersive x-ray spectroscopy (EDX) of soil, dust, and rock particles in the field. The MEMSA would be similar to an environmental scanning electron microscope (ESEM) but would be much smaller and designed specifically for field use in studying effects of geological alteration at the micrometer scale. Like an ESEM, the MEMSA could be used to examine uncoated, electrically nonconductive specimens. In addition to the difference in size, other significant differences between the MEMSA and an ESEM lie in the mode of scanning and the nature of the electron source.

  16. Stability Studies on Dry Swabs and Wet Mailed Swabs for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Aptima Assays.

    Science.gov (United States)

    Moncada, Jeanne; Clark, Carey B; Holden, Jeffrey; Hook, Edward W; Gaydos, Charlotte A; Schachter, Julius

    2017-03-01

    The Aptima Combo 2 (AC2) and Aptima CT (ACT) (Hologic Inc., San Diego, CA) are nucleic acid amplification tests (NAATs) that detect Chlamydia trachomatis AC2 also detects Neisseria gonorrhoeae Storage and temperature conditions may impact the utility of NAATs in some settings and screening programs. We evaluated specimen stability for use beyond the Aptima package insert specifications for temperature and duration of storage (between 2°C and 30°C and 60 days, respectively) in two studies: (i) dry C. trachomatis-seeded swabs were used with ACT after storage at 4°C, 23°C, or 36°C for up to 84 days and (ii) swabs seeded with C. trachomatis and N. gonorrhoeae and then placed in transport medium were tested with AC2, after being mailed via the U.S. Postal Service to three different sites. Prolonged storage of samples had no effect, and samples stored at 4°C, 23°C, and 36°C for up to 84 days yielded comparable ACT positivities, although there was a drop in signal intensity for virtually all specimens under all storage/shipping conditions after day 21. In the mailing study, 80%, 52% and 29% of seeded swabs were exposed to temperatures of >30°C during three rounds in transit, and 2% reached temperatures of >40°C. No evidence of signal degradation in the AC2 assay for detection of C. trachomatis or N. gonorrhoeae was observed, although some mailed swabs took more than 5 weeks to reach the laboratory site. These two studies support the potential use of swabs at temperatures above 36°C and storage beyond 60 days and provide confidence regarding this commercially available NAAT for testing of specimens after mailing. Copyright © 2017 American Society for Microbiology.

  17. A Proposed Typology of Sampled Material Within Electronic Dance Music

    Directory of Open Access Journals (Sweden)

    Robert Ratcliffe

    2014-06-01

    Full Text Available The following article contains a proposed typology of sampled material within electronic dance music (EDM. The typology offers a system of classification that takes into account the sonic, musical and referential properties of sampled elements, while also considering the technical realisation of material and the compositional intentions of the producer/DJ. Illustrated with supporting examples drawn from a wide variety of artists and sub-genres, the article seeks to address the current lack of research on the subject of sample-based composition and production, and provide a theoretical framework for further discussion of EDM sampling practices. In addition, it demonstrates how concepts and terminology derived from the field of electroacoustic music may be successfully applied to the study and analysis of EDM, resulting in an expanded analytical methodology.

  18. Comparison of feces versus rectal swabs for the molecular detection of Lawsonia intracellularis in foals with equine proliferative enteropathy.

    Science.gov (United States)

    Pusterla, Nicola; Mapes, Samantha; Johnson, Cara; Slovis, Nathan; Page, Allen; Gebhart, Connie

    2010-09-01

    The purpose of the current study was to compare the molecular detection rate of Lawsonia intracellularis between feces and rectal swabs collected from 42 foals with suspected equine proliferative enteropathy (EPE). Fecal samples and rectal swabs were processed for DNA purification by using an automated extraction system. The purified DNA was then analyzed by real-time polymerase chain reaction (PCR) for the presence of the aspartate ammonia lyase (aspA) gene of L. intracellularis. Absolute quantitation was calculated by using a standard curve for L. intracellularis and expressed as copy numbers of the aspA gene of L. intracellularis per microliter of purified DNA. The combined PCR detection rate for L. intracellularis was 90%, with 38 foals testing PCR positive in feces (33 samples), rectal swabs (32), or both (27). Six foals tested PCR positive only in feces, whereas 5 tested positive only in rectal swabs. Feces yielded a significantly higher aspA gene copy number of L. intracellularis than rectal swabs. Feces and rectal swabs tested PCR negative from 4 foals. In conclusion, the results showed that feces yielded similar numbers of PCR-positive results, with a higher L. intracellularis aspA gene load than rectal swabs. By analyzing dual samples, the PCR detection rate for L. intracellularis increased from 76% and 79% for rectal swabs and feces, respectively, to 90%. Rectal swabs should be considered as an alternative sample type for EPE-suspected patients with decreased or no fecal output.

  19. Evaluation of the Recovery Rate of Different Swabs for Microbial Environmental Monitoring.

    Science.gov (United States)

    Goverde, Marcel; Willrodt, Julian; Staerk, Alexandra

    realistic and might better reflect what is found in pharmaceutical clean rooms. Environmental microbiological monitoring provides information on the hygiene condition of pharmaceutical clean rooms and equipment for manufacturing of drug products. Different methods can be used to recover microorganisms. For surfaces, normally contact plates (e.g., RODAC or dipslides) are used; however, when surfaces are uneven, swabs should be used. In the present study three different swabs were evaluated for their ability to recover microorganisms from different surfaces. Thereby two methods and two approaches were evaluated. Swab samples were either directly stroked on agar or the swab was eluted, membrane-filtrated, and the filter placed on an agar plate. Experimentally, artificial inoculated surfaces typically found in clean rooms (in vitro study) and naturally inoculated floors (in situ study) were sampled. Thus with this approach the most convenient swab and the most suitable recovery methods under laboratory as well as real clean room conditions were evaluated. With this set-up, we found the most suitable swab for our environmental monitoring not only by using artificial inoculated surfaces but also under more realistic clean room conditions, which is most important for microbiological environmental monitoring sampling. © PDA, Inc. 2017.

  20. Microbial contamination assessment of cryostored autogenous cranial bone flaps: should bone biopsies or swabs be performed?

    Science.gov (United States)

    Bhaskar, Ivan P; Inglis, Timothy J J; Bowman, Jacintha; Lee, Gabriel Y F

    2013-02-01

    Autogenous cranioplasty infection requiring bone flap removal is under-recognised as a major complication causing significant morbidity. Microbial contamination of stored bone flaps may be a significant contributing factor. Current infection control practices and storage procedures vary. It is not known whether 'superficial' swabs or bone cultures provide a more accurate assessment. Twenty-five skull flaps that were cryo-stored for more than 6 months were studied. Two swab samples (superficial and deep) and a bone biopsy sample were taken from each skull flap sample and cultured. Half blood agar and half chocolate agar plates were inoculated with the swabs for anaerobic and aerobic cultures respectively. The bone biopsy samples were cultured in brain-heart broth and subcultured similar to the swabs for 5 days. Incidence of microbial contamination was 20 % in the bone flaps studied. One swab culture and five bone biopsy cultures were positive for bacterial growth, all of which contained Propionibacterium acnes (p = 0.014). Positive cultures were from bone flaps stored less than 18 months, whereas no growth was obtained from bone flaps that were stored longer (p = 0.014). Bone biopsy culture is a more sensitive technique of assessing microbial contamination of cryo-stored autogenous bone flaps than swab cultures. The clinical implications of in vitro demonstration of microbial contamination require further study.

  1. Evaluation of the Swabbing of Disposable Absorbent Incontinence Products for Assessing the Carriage of Multiresistant Enterobacteriaceae in Nursing Home Residents

    Directory of Open Access Journals (Sweden)

    Alexis Naf

    2017-09-01

    Full Text Available We compared the performance of incontinence product (IP and rectal swabbing for the detection of multidrug-resistant Enterobacteriaceae (MDRE carriage in a large multicenter study conducted in February 2017 among the residents of 23 French nursing homes. The study included 547 residents who habitually wore IP, 88 of whom were MDRE carriers (16.1%. Positive results were obtained for both rectal and IP swabs for 64 of these residents, for rectal swabs only for 22 and for IP swabs only for two of these patients. The estimated prevalence of MDRE carriage depended on the type of sample: 15.7% for rectal swabs and 12.1% for IP swabs (p < 0.001. The positive percent agreement was 84.2% and the negative percent agreement was 97.4%. Rectal swabbing remains the best method for detecting MDRE carriage in elderly residents, but our findings provide support for the use of swabs from IP used overnight to increase response rates in MDRE surveys in elderly residents that habitually wear IP, when rectal swabbing is not feasible.

  2. Self-collected mid-turbinate swabs for the detection of respiratory viruses in adults with acute respiratory illnesses.

    Directory of Open Access Journals (Sweden)

    Oscar E Larios

    Full Text Available BACKGROUND: The gold standard for respiratory virus testing is a nasopharyngeal (NP swab, which is collected by a healthcare worker. Midturbinate (MT swabs are an alternative due to their ease of collection and possible self-collection by patients. The objective of this study was to compare the respiratory virus isolation of flocked MT swabs compared to flocked NP swabs. METHODS: Beginning in October 2008, healthy adults aged 18 to 69 years were recruited into a cohort and followed up for symptoms of influenza. They were asked to have NP and MT swabs taken as soon as possible after the onset of a fever or two or more respiratory symptoms with an acute onset. The swabs were tested for viral respiratory infections using Seeplex® RV12 multiplex PCR detection kit. Seventy six pairs of simultaneous NP and MT swabs were collected from 38 symptomatic subjects. Twenty nine (38% of these pairs were positive by either NP or MT swabs or both. Sixty nine (91% of the pair results were concordant. Two samples (3% for hCV OC43/HKU1 and 1 sample (1% for rhinovirus A/B were positive by NP but negative by MT. One sample each for hCV 229E/NL63, hCV OC43/HKU1, respiratory syncytial virus A, and influenza B were positive by MT but negative by NP. CONCLUSIONS: Flocked MT swabs are sensitive for the diagnosis of multiple respiratory viruses. Given the ease of MT collection and similar results between the two swabs, it is likely that MT swabs should be the preferred method of respiratory cell collection for outpatient studies. In light of this data, larger studies should be performed to ensure that this still holds true and data should also be collected on the patient preference of collection methods.

  3. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Lee, Soomin; Nishimura, Takayuki; Katsuura, Tetsuo; Yasukouchi, Akira

    2011-01-10

    Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO). In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). The melatonin levels were analyzed in duplicate using commercially available ELISA kits. The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL). The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level (<6 pg/mL), although the BA plots didn't show proportional and relative biases, there was no significant correlation between passive and cotton saliva collection samples. Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  4. Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy.

    Science.gov (United States)

    Can, Hüseyin; Caner, Ayşe; Döşkaya, Mert; Değirmenci, Aysu; Karaçalı, Sabire; Polat, Ceylan; Gürüz, Yüksel; Uner, Ahmet

    2013-02-13

    Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone. Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration of dexamethasone. Also, samples were collected from lung, heart, liver, kidney, diaphragm, brain, spleen, tongue, muscle, eye, intestine, and feces. Cysts and trophozoites were investigated in stained slides and MSG gene was detected by PCR. The results showed that weight loss is significantly higher in rats administered oral dexamethasone (Pdexamethasone. PCR was positive in lungs and oral swabs of rats prior to the administration of dexamethasone. After the administration of dexamethasone, the MSG gene was detected in oral swabs, lungs, spleen, kidney and (for the first time) in nasal swabs. PCR was positive in nasal swabs during the second and sixth weeks of oral and subcutaneous administration of dexamethasone, respectively. Presence of P. jiroveci in nasopharyngeal aspirate, oropharyngeal wash, oral swab, induced sputum or BAL, and absence in nasal swab in a patient without symptoms of PCP may support clinician's decision regarding colonization. Overall, detection of P. carinii in nasal swabs of rats by PCR demonstrated that nasal sampling can be used for the diagnosis of Pneumocystis pneumonia.

  5. Cheek swabs, SNP chips, and CNVs: Assessing the quality of copy number variant calls generated with subject-collected mail-in buccal brush DNA samples on a high-density genotyping microarray

    Directory of Open Access Journals (Sweden)

    Erickson Stephen W

    2012-06-01

    Full Text Available Abstract Background Multiple investigators have established the feasibility of using buccal brush samples to genotype single nucleotide polymorphisms (SNPs with high-density genome-wide microarrays, but there is currently no consensus on the accuracy of copy number variants (CNVs inferred from these data. Regardless of the source of DNA, it is more difficult to detect CNVs than to genotype SNPs using these microarrays, and it therefore remains an open question whether buccal brush samples provide enough high-quality DNA for this purpose. Methods To demonstrate the quality of CNV calls generated from DNA extracted from buccal samples, compared to calls generated from blood samples, we evaluated the concordance of calls from individuals who provided both sample types. The Illumina Human660W-Quad BeadChip was used to determine SNPs and CNVs of 39 Arkansas participants in the National Birth Defects Prevention Study (NBDPS, including 16 mother-infant dyads, who provided both whole blood and buccal brush DNA samples. Results We observed a 99.9% concordance rate of SNP calls in the 39 blood–buccal pairs. From the same dataset, we performed a similar analysis of CNVs. Each of the 78 samples was independently segmented into regions of like copy number using the Optimal Segmentation algorithm of Golden Helix SNP & Variation Suite 7. Across 640,663 loci on 22 autosomal chromosomes, segment-mean log R ratios had an average correlation of 0.899 between blood-buccal pairs of samples from the same individual, while the average correlation between all possible blood-buccal pairs of samples from unrelated individuals was 0.318. An independent analysis using the QuantiSNP algorithm produced average correlations of 0.943 between blood-buccal pairs from the same individual versus 0.332 between samples from unrelated individuals. Segment-mean log R ratios had an average correlation of 0.539 between mother-offspring dyads of buccal samples, which was not

  6. Effects of saliva collection using cotton swabs on melatonin enzyme immunoassay

    Directory of Open Access Journals (Sweden)

    Katsuura Tetsuo

    2011-01-01

    Full Text Available Abstract Background Although various acceptable and easy-to-use devices have been used for saliva collection, cotton swabs are among the most common ones. Previous studies reported that cotton swabs yield a lower level of melatonin detection. However, this statistical method is not adequate for detecting an agreement between cotton saliva collection and passive saliva collection, and a test for bias is needed. Furthermore, the effects of cotton swabs have not been examined at lower melatonin level, a level at which melatonin is used for assessment of circadian rhythms, namely dim light melatonin onset (DLMO. In the present study, we estimated the effect of cotton swabs on the results of salivary melatonin assay using the Bland-Altman plot at lower level. Methods Nine healthy males were recruited and each provided four saliva samples on a single day to yield a total of 36 samples. Saliva samples were directly collected in plastic tubes using plastic straws, and subsequently pipetted onto cotton swabs (cotton saliva collection and into clear sterile tubes (passive saliva collection. The melatonin levels were analyzed in duplicate using commercially available ELISA kits. Results The mean melatonin concentration in cotton saliva collection samples was significantly lower than that in passive saliva collection samples at higher melatonin level (>6 pg/mL. The Bland-Altman plot indicated that cotton swabs causes relative and proportional biases in the assay results. For lower melatonin level ( Conclusion Our findings indicate an interference effect of cotton swabs on the assay result of salivary melatonin at lower melatonin level. Cotton-based collection devices might, thus, not be suitable for assessment of DLMO.

  7. Evaluation of methods to improve the extraction and recovery of DNA from cotton swabs for forensic analysis.

    Science.gov (United States)

    Adamowicz, Michael S; Stasulli, Dominique M; Sobestanovich, Emily M; Bille, Todd W

    2014-01-01

    Samples for forensic DNA analysis are often collected from a wide variety of objects using cotton or nylon tipped swabs. Testing has shown that significant quantities of DNA are retained on the swab, however, and subsequently lost. When processing evidentiary samples, the recovery of the maximum amount of available DNA is critical, potentially dictating whether a usable profile can be derived from a piece of evidence or not. The QIAamp DNA Investigator extraction kit was used with its recommended protocol for swabs (one hour incubation at 56°C) as a baseline. Results indicate that over 50% of the recoverable DNA may be retained on the cotton swab tip, or otherwise lost, for both blood and buccal cell samples when using this protocol. The protocol's incubation time and temperature were altered, as was incubating while shaking or stationary to test for increases in recovery efficiency. An additional step was then tested that included periodic re-suspension of the swab tip in the extraction buffer during incubation. Aliquots of liquid blood or a buccal cell suspension were deposited and dried on cotton swabs and compared with swab-less controls. The concentration of DNA in each extract was quantified and STR analysis was performed to assess the quality of the extracted DNA. Stationary incubations and those performed at 65°C did not result in significant gains in DNA yield. Samples incubated for 24 hours yielded less DNA. Increased yields were observed with three and 18 hour incubation periods. Increases in DNA yields were also observed using a swab re-suspension method for both cell types. The swab re-suspension method yielded an average two-fold increase in recovered DNA yield with buccal cells and an average three-fold increase with blood cells. These findings demonstrate that more of the DNA collected on swabs can be recovered with specific protocol alterations.

  8. Quantification of loosely associated and tightly associated bacteria on broiler carcass skin using swabbing, stomaching, and grinding methods.

    Science.gov (United States)

    Singh, P; Lee, H C; Chin, K B; Ha, S D; Kang, I

    2015-12-01

    This research was conducted to quantify bacterial populations after swabbing or stomaching, followed by grinding the swabbed or stomached broiler skins. For each of 3 replications, 3 eviscerated broilers were randomly taken from a processing line in a local broiler processing plant. Ten swabs and 10 stomachs per bird were conducted on the left- and the right-side skins (10×7 cm), respectively, which were then finally ground. Results indicated that mesophilic aerobic bacteria (MAB) in the first swabbed sample were significantly lower than those in the first stomached sample (P0.05). During 10 swabbings followed by final grinding, 8, 9, and 83% of MAB were detected after the first swabbing, after the second through 10th swabbings, and after final grinding of the skin, respectively. During 10 stomachings followed by the final grinding, 17, 18, and 65% of MAB were detected after the first stomaching, after the second through 10th stomachings, and after final grinding of the skin, respectively. Escherichia coli (E. coli) and coliforms were significantly higher in the first stomaching than those in the first swabbing (P0.05). Populations of E. coli and coliforms decreased step-wisely from the highest after grinding to the intermediate after first and second sampling, and to the least after 10th sampling (P<0.05), regardless of swabbing or grinding. In this study, less than 35% of MAB seemed loosely associated in the skin of eviscerated broiler, whereas more than 65% of MAB looked tightly associated, which were not recovered by stomaching or swabbing even 10 times but were recovered by grinding the skin. © 2015 Poultry Science Association Inc.

  9. Evaluation of Methods to Improve the Extraction and Recovery of DNA from Cotton Swabs for Forensic Analysis

    Science.gov (United States)

    Adamowicz, Michael S.; Stasulli, Dominique M.; Sobestanovich, Emily M.; Bille, Todd W.

    2014-01-01

    Samples for forensic DNA analysis are often collected from a wide variety of objects using cotton or nylon tipped swabs. Testing has shown that significant quantities of DNA are retained on the swab, however, and subsequently lost. When processing evidentiary samples, the recovery of the maximum amount of available DNA is critical, potentially dictating whether a usable profile can be derived from a piece of evidence or not. The QIAamp DNA Investigator extraction kit was used with its recommended protocol for swabs (one hour incubation at 56°C) as a baseline. Results indicate that over 50% of the recoverable DNA may be retained on the cotton swab tip, or otherwise lost, for both blood and buccal cell samples when using this protocol. The protocol’s incubation time and temperature were altered, as was incubating while shaking or stationary to test for increases in recovery efficiency. An additional step was then tested that included periodic re-suspension of the swab tip in the extraction buffer during incubation. Aliquots of liquid blood or a buccal cell suspension were deposited and dried on cotton swabs and compared with swab-less controls. The concentration of DNA in each extract was quantified and STR analysis was performed to assess the quality of the extracted DNA. Stationary incubations and those performed at 65°C did not result in significant gains in DNA yield. Samples incubated for 24 hours yielded less DNA. Increased yields were observed with three and 18 hour incubation periods. Increases in DNA yields were also observed using a swab re-suspension method for both cell types. The swab re-suspension method yielded an average two-fold increase in recovered DNA yield with buccal cells and an average three-fold increase with blood cells. These findings demonstrate that more of the DNA collected on swabs can be recovered with specific protocol alterations. PMID:25549111

  10. Throat Swabs and Sputum Culture as Predictors of P. aeruginosa or S. aureus Lung Colonization in Adult Cystic Fibrosis Patients.

    Directory of Open Access Journals (Sweden)

    Darius Seidler

    Full Text Available Due to frequent infections in cystic fibrosis (CF patients, repeated respiratory cultures are obtained to inform treatment. When patients are unable to expectorate sputum, clinicians obtain throat swabs as a surrogate for lower respiratory cultures. There is no clear data in adult subjects demonstrating the adequacy of throat swabs as a surrogate for sputum or BAL. Our study was designed to determine the utility of throat swabs in identifying lung colonization with common organisms in adults with CF.Adult CF subjects (n = 20 underwent bronchoscopy with BAL. Prior to bronchoscopy, a throat swab was obtained. A sputum sample was obtained from subjects who were able to spontaneously expectorate. All samples were sent for standard microbiology culture.Using BAL as the gold standard, we found the positive predictive value for Pseudomonas aeruginosa to be 100% in both sputum and throat swab compared to BAL. However, the negative predictive value for P. aeruginosa was 60% and 50% in sputum and throat swab, respectively. Conversely, the positive predictive value for Staphylococcus aureus was 57% in sputum and only 41% in throat swab and the negative predictive value of S. aureus was 100% in sputum and throat swab compared to BAL.Our data show that positive sputum and throat culture findings of P. aeruginosa reflect results found on BAL fluid analysis, suggesting these are reasonable surrogates to determine lung colonization with P. aeruginosa. However, sputum and throat culture findings of S. aureus do not appear to reflect S. aureus colonization of the lung.

  11. Fabrication of SERS swab for direct detection of trace explosives in fingerprints.

    Science.gov (United States)

    Gong, Zhengjun; Du, Hongjie; Cheng, Fansheng; Wang, Cong; Wang, Canchen; Fan, Meikun

    2014-12-24

    Swab sampling is of great importance in surface contamination analysis. A cotton swab (cotton Q-tip) was successfully transformed into surface-enhanced Raman scattering (SERS) substrate (SERS Q-tip) through a bottom-up strategy, where Ag NPs were first self-assembled onto the Q-tip followed by in situ growing. The capability for direct swab detection of Raman probe Nile Blue A (NBA) and a primary explosive marker 2,4-dinitrotoluene (2,4-DNT) using the SERS Q-tip was explored. It was found that at optimum conditions, a femotogram of NBA on glass surface could be swab-detected. The lowest detectable amount for 2,4-DNT is only ∼1.2 ng/cm(2) (total amount of 5 ng) on glass surface, 2 orders of magnitude more sensitive than similar surface analysis achieved with infrared technique, and comparable even with that obtained by ion mobility spectrometry-mass spectrometry. Finally, 2,4-DNT left on fingerprints was also analyzed. It was found that SERS signal of 2,4-DNT from 27th fingerprint after touching 2,4-DNT powder can still be clearly identified by swabbing with the SERS Q-tip. We believe this is the first direct SERS swabbing test of explosives on fingerprint on glass. Considering its relative long shelf life (>30 d), the SERS Q-tip may find great potential in future homeland security applications when combined with portable Raman spectrometers.

  12. Electronic paramagnetic resonance power saturation of wooden samples

    Science.gov (United States)

    Brai, Maria; Longo, Anna; Maccotta, Antonella; Marrale, Maurizio

    2009-05-01

    The deterioration of wood used for artifacts of artistic interest involves the production of different free radicals from the macromolecules of the wooden matrix (cellulose, lignin, and hemicellulose). Among the techniques able to provide information about these free radicals, the contribution of electronic paramagnetic resonance (EPR) can be very valuable. In this paper, the study of EPR signals (with g ≈2) of both modern and ancient wooden taxa was undertaken in order to analyze some features of the free radicals in natural wood. In particular, we have studied the microwave power saturation behaviors of seasoned wooden samples from ten species, and we have found remarkable differences between softwoods and hardwoods. These differences can be correlated to dissimilarities in the relaxation times T1 and T2 attributable to the different microscopic structures of the two trees' categories. The method has been also applied to ancient woods belonging to works of art in order to assess the conservation state of these artifacts. The analysis of the saturation curves has been found to be sensitive to the wood decay state. Indeed the deterioration process of the wooden matrix involves a variation of the relaxation times; this could be ascribed to both possible structure modifications and to concentration increments of the free radicals inside ancient woods due to decay induced by natural (biological, chemical, and physical) agents. This analysis method seems to be promising for the characterization of the wooden decay state and, therefore, it could provide valuable diagnostic indications which are necessary for the restoration and conservation of many artifact of historical-artistic-archaeological interest.

  13. Detection of Brucella spp. in bottlenose dolphins Tursiops truncatus by a real-time PCR using blowhole swabs.

    Science.gov (United States)

    Wu, Qingzhong; Conway, Jessica; Phillips, Kristen M; Stolen, Megan; Durden, Wendy N; Fauquier, Deborah; McFee, Wayne E; Schwacke, Lori

    2016-08-09

    Blowhole swabs are a simple and non-invasive method for collecting samples from cetaceans and can be used for screening large numbers of animals in the field. This study reports a real-time PCR assay for the detection of Brucella spp. using blowhole swab samples from bottlenose dolphins Tursiops truncatus stranded in the coastal region of Virginia, South Carolina and northern Florida, USA, between 2013 and 2015. We used real-time PCR results on lung samples from the same dolphins in order to estimate the relative sensitivity and specificity of real-time PCR of blowhole swabs. Brucella DNA was detected in lung tissue of 22% (18/81) and in blowhole swabs of 21% (17/81) of the sampled dolphins. The relative sensitivity and specificity of real-time PCR on blowhole swabs as compared to the real-time PCR on lung samples was 94% (17/18) and 100% (63/63), respectively. These results indicate that real-time PCR on blowhole swabs may be used as a non-invasive test for rapid detection of Brucella spp. in the respiratory tract of dolphins. To our knowledge, this is the first report on the use of blowhole swabs for detection of bacterial pathogens by real-time PCR in bottlenose dolphins.

  14. Arrangement and method for accommodating a sample in an electron microscope

    OpenAIRE

    H.W. Zandbergen

    1996-01-01

    Abstract of NL 9402226 (A) The invention relates to an arrangement for accommodating a sample in an electron microscope, equipped with a sample holder which comprises sample accommodation means and is designed for at least partial accommodation in an electron microscope, where at least one sample mount is provided which can contain a sample in a position in which the sample is at least partially visible from the outside of the sample mount from at least two sides opposite one another, the or ...

  15. Performance of self-collected penile-meatal swabs compared to clinician-collected urethral swabs for the detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, and Mycoplasma genitalium by nucleic acid amplification assays.

    Science.gov (United States)

    Dize, Laura; Barnes, Perry; Barnes, Mathilda; Hsieh, Yu-Hsiang; Marsiglia, Vincent; Duncan, Della; Hardick, Justin; Gaydos, Charlotte A

    2016-10-01

    Men were enrolled in a study to assess the performance and acceptability of self-collected penile meatal swabs as compared to clinician-collected urethral swabs for sexually transmitted infections (STIs). We expected penile-meatal swabs to perform favorably to urethral swabs for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Trichomonas vaginalis (TV), and Mycoplasma genitalium (MG) detection by nucleic acid amplification assays (NAATs). Of 203 swab pairs tested; for CT, penile-meatal swab sensitivity was 96.8% and specificity was 98.8%. NG sensitivity and specificity were 100% and 98.9%, respectively. For TV, sensitivity was 85.0% and specificity was 96.7%. For MG sensitivity and specificity were 79.3% and 99.4%, respectively. No significant statistical differences between sample type accuracy (CT: P=0.625; NG: P=0.248; TV: P=0.344; and MG: P=0.070) existed. Most men, 90.1%, reported self-collection of penile-meatal swabs as "Very Easy" or "Easy". Self-collected penile-meatal swabs appeared acceptable for NAAT STI detection and an acceptable collection method by men. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Arrangement and method for accommodating a sample in an electron microscope

    NARCIS (Netherlands)

    Zandbergen, H.W.

    1996-01-01

    Abstract of NL 9402226 (A) The invention relates to an arrangement for accommodating a sample in an electron microscope, equipped with a sample holder which comprises sample accommodation means and is designed for at least partial accommodation in an electron microscope, where at least one sample

  17. Effects of saliva collection using cotton swab on cortisol enzyme immunoassay.

    Science.gov (United States)

    Kozaki, Tomoaki; Hashiguchi, Nobuko; Kaji, Yumi; Yasukouchi, Akira; Tochihara, Yutaka

    2009-12-01

    Cotton swabs are among the most commonly used devices for collecting saliva, but various studies have reported that their use impacts the results of salivary cortisol assays. These studies, however, estimated this impact by comparing the average of the concentration and/or scatter plots. In the present study, we estimated the impact of cotton swabs on the results of salivary cortisol enzyme immunoassay (EIA) by Bland-Altman plot. Eight healthy males (aged 20-23 years) provided four saliva samples on different days to yield a total of 32 samples. Saliva samples were collected directly in plastic tubes using plastic straws and then pipetted onto cotton swabs (cotton saliva collection) and into clear sterile tubes (passive saliva collection). There was a lower correlation between cotton and passive saliva collection. Individually, four subjects showed a negative correlation between passive and cotton saliva collection. A Bland-Altman plot indicated that cotton swabs causes a proportional bias on the EIA assay result. Our findings indicate a considerable effect of using cotton swabs for saliva collection, and subject-specific variability in the impact. A Bland-Altman plot further suggests possible reasons for this effect.

  18. Sparse sampling and reconstruction for electron and scanning probe microscope imaging

    Science.gov (United States)

    Anderson, Hyrum; Helms, Jovana; Wheeler, Jason W.; Larson, Kurt W.; Rohrer, Brandon R.

    2015-07-28

    Systems and methods for conducting electron or scanning probe microscopy are provided herein. In a general embodiment, the systems and methods for conducting electron or scanning probe microscopy with an undersampled data set include: driving an electron beam or probe to scan across a sample and visit a subset of pixel locations of the sample that are randomly or pseudo-randomly designated; determining actual pixel locations on the sample that are visited by the electron beam or probe; and processing data collected by detectors from the visits of the electron beam or probe at the actual pixel locations and recovering a reconstructed image of the sample.

  19. Culture-independent qunatification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2010-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation......, for culture-independent quantification of intact Salmonella in pig carcass gauze swabs (100 cm2) prior to quantitative PCR (qPCR). A novel approach was investigated, excluding the homogenization step prior to flotation, to improve the detection limit and speed up the quantification procedure. The buoyant...... concentrations of ≥ 6.1×108 CFU/swab sample, but not by concentrations ≤ 6.1×106 CFU/swab sample. By using the gauze swabs directly in the flotation procedure, the homogenization step normally used for preparation of food-related samples could be excluded, which simplified the culture independent quantification...

  20. Validation of Single and Pooled Manure Drag Swabs for the Detection of Salmonella Serovar Enteritidis in Commercial Poultry Houses.

    Science.gov (United States)

    Kinde, Hailu; Goodluck, Helen A; Pitesky, Maurice; Friend, Tom D; Campbell, James A; Hill, Ashley E

    2015-12-01

    Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n  =  525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯  = 2.5 CFU (range 2.5-2.7), or medium, x¯  = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P 0.05) between the FDA method (single swabs) and the pooled NPIP method (four-pool swabs). The study concludes that the pooled NPIP method is not significantly different from the FDA method for the detection of Salmonella Enteritidis in drag swabs in commercial poultry laying houses. Consequently based on the FDA

  1. Does Fine Needle Aspiration Microbiology Offer Any Benefit Over Wound Swab in Detecting the Causative Organisms in Surgical Site Infections?

    Science.gov (United States)

    Sudharsanan, Sundaramurthi; Gs, Sreenath; Sureshkumar, Sathasivam; Vijayakumar, Chellappa; Sujatha, Sistla; Kate, Vikram

    2017-09-01

    The objective of this study is to determine the role of ne needle aspiration microbiology (FNAM) in detecting the causative organisms of postoperative surgical site infections (SSIs) in comparison with the standard technique of surface swabbing. Ma- terials and Methods. In this study, 150 patients with SSIs following elective and emergency operations were included. In all patients, FNAM was performed along with conventional surface swabbing to identify the causative microorganism. Sensitivity of surface swab and FNAM was calculated as the number of samples collected from the diagnosed case of SSI. A total of 115 positive cultures were obtained from the 150 patients with SSIs; surface swab was positive in 110 cases and FNAM was positive in 94 cases. The mean number of organisms isolated by surface swab, and FNAM was 0.95 and 0.8, respectively. The sensitivity of surface swab was 94.3% in elective cases and 96.25% in emergency cases. The sensitivity of FNAM was 82.8% in elective cases and 82.5% in emergency cases. The sensitivity and negative predictive value of FNAM and surface swab did not signi cantly differ in clean elective cases. The overall sensitivity of surface swab and FNAM was 95.65% and 81.7%, respectively. Comparing the antibiotic suscep- tibility pattern, no difference was observed when the same organ- ism was isolated by both methods, indicating that FNAM does not offer bene t over the conventional wound surface swab in detecting microorganisms in SSI in both elective and emergency surgeries. In certain cases with unexplained wound infections, FNAM can be used as an investigation to identify speci c pathogens not detected by conventional surface swab.

  2. Validation of a Nylon-Flocked-Swab Protocol for Efficient Recovery of Bacterial Spores from Smooth and Rough Surfaces▿

    Science.gov (United States)

    Probst, Alexander; Facius, Rainer; Wirth, Reinhard; Moissl-Eichinger, Christine

    2010-01-01

    In order to meet planetary-protection requirements, culturable bacterial spore loads are measured representatively for the total microbial contamination of spacecraft. However, the National Aeronautics and Space Administration's (NASA's) cotton swab protocols for spore load determination have not changed for decades. To determine whether a more efficient alternative was available, a novel swab was evaluated for recovery of different Bacillus atrophaeus spore concentrations on stainless steel and other surfaces. Two protocols for the nylon-flocked swab (NFS) were validated and compared to the present NASA standard protocol. The results indicate that the novel swab protocols recover 3- to 4-fold more (45.4% and 49.0% recovery efficiency) B. atrophaeus spores than the NASA standard method (13.2%). Moreover, the nylon-flocked-swab protocols were superior in recovery efficiency for spores of seven different Bacillus species, including Bacillus anthracis Sterne (recovery efficiency, 20%). The recovery efficiencies for B. atrophaeus spores from different surfaces showed a variation from 5.9 to 62.0%, depending on the roughness of the surface analyzed. Direct inoculation of the swab resulted in a recovery rate of about 80%, consistent with the results of scanning electron micrographs that allowed detailed comparisons of the two swab types. The results of this investigation will significantly contribute to the cleanliness control of future life detection missions and will provide significant improvement in detection of B. anthracis contamination for law enforcement and security efforts. PMID:20543054

  3. Comparison of sputum and nasopharyngeal swabs for detection of respiratory viruses.

    Science.gov (United States)

    Jeong, Ji Hun; Kim, Kyung Hee; Jeong, Sung Hwan; Park, Jeong Woong; Lee, Sang Min; Seo, Yiel Hea

    2014-12-01

    Diagnostic tests for respiratory viral infections use traditionally either nasopharyngeal washes or swabs. Sputum is representative of the lower respiratory tract but is used rarely for viral testing. The aim of this study was to compare the detection rates of respiratory viruses from nasopharyngeal swabs and sputum using a multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR). Adults who were admitted or presented to the clinics of Gil Medical Center with acute respiratory symptoms were recruited from 1 November 2012 to 31 March 2013. Paired specimens of nasopharyngeal swabs and sputum were obtained from 154 subjects, and RNA was extracted and tested for 16 different respiratory viruses using the Anyplex II RV16 Detection kit (Seegene, Seoul, Korea). The positive rate was 53% (81/154) for nasopharyngeal swabs and 68% (105/154) for sputum (P < 0.001). One hundred thirty-four viruses were identified for 107 illnesses. Influenza A virus, RSV A, HRV, coronavirus OC43, and adenovirus were detected more frequently in sputum samples than in nasopharyngeal swabs (P < 0.001). Importantly, 12 of 44 (27%) influenza A infections and 11 of 27 (41%) RSV infections were positive in only sputum samples. The detection rates of respiratory viruses from sputum samples were significantly higher than those from nasopharyngeal swabs in adults using real-time multiplex RT-PCR. These findings suggest that sputum would benefit for the detection of respiratory viruses by nucleic acid amplification tests (NAATs) in patients who produce sputum. Further studies are needed to establish standardized RNA extraction methods from sputum samples. © 2014 Wiley Periodicals, Inc.

  4. Surface, Water, and Air Biocharacterization (SWAB) Flight Experiment

    Science.gov (United States)

    Castro, V. A.; Ott, C. M.; Pierson, D. L.

    2012-01-01

    The determination of risk from infectious disease during spaceflight missions is composed of several factors including both the concentration and characteristics of the microorganisms to which the crew are exposed. Thus, having a good understanding of the microbial ecology aboard spacecraft provides the necessary information to mitigate health risks to the crew. While preventive measures are taken to minimize the presence of pathogens on spacecraft, medically significant organisms have been isolated from both the Mir and International Space Station (ISS). Historically, the method for isolation and identification of microorganisms from spacecraft environmental samples depended upon their growth on culture media. Unfortunately, only a fraction of the organisms may grow on a specific culture medium, potentially omitting those microorganisms whose nutritional and physical requirements for growth are not met. To address this bias in our understanding of the ISS environment, the Surface, Water, and Air Biocharacterization (SWAB) Flight Experiment was designed to investigate and develop monitoring technology to provide better microbial characterization. For the SWAB flight experiment, we hypothesized that environmental analysis using non-culture-based technologies would reveal microorganisms, allergens, and microbial toxins not previously reported in spacecraft, allowing for a more complete health assessment. Key findings during this experiment included: a) Generally, advanced molecular techniques were able to reveal a few organisms not recovered using culture-based methods; however, there is no indication that current monitoring is "missing" any medically significant bacteria or fungi. b) Molecular techniques have tremendous potential for microbial monitoring, however, sample preparation and data analysis present challenges for spaceflight hardware. c) Analytical results indicate that some molecular techniques, such as denaturing gradient gel electrophoresis (DGGE), can

  5. Comparing non-woven, filmateci and woven gauze swabs.

    Science.gov (United States)

    Thomas, S; Loveless, P; Hay, N P; Toyick, N

    1993-01-02

    The physical characteristics and performance of seven non-woven swabs intended for topical use were compared with those of filmated swabs and woven cotton gauze in a series of laboratory tests. The results of this study suggest that the non-woven swabs have significant advantages over the other type examined. Based upon current pricing structures they represent a highly cost-effective alternative to the more traditional products for routine wound management procedures. As the various non-wovens have very different handling characteristics, it should be possible to select a swab to suit most requirements from the range of products available.

  6. Transmission electron microscope cells for use with liquid samples

    Energy Technology Data Exchange (ETDEWEB)

    Khalid, Waqas; Alivisatos, Paul A.; Zettl, Alexander K.

    2016-08-09

    This disclosure provides systems, methods, and devices related to transmission electron microscopy cells for use with liquids. In one aspect a device includes a substrate, a first graphene layer, and a second graphene layer. The substrate has a first surface and a second surface. The first surface defines a first channel, a second channel, and an outlet channel. The first channel and the second channel are joined to the outlet channel. The outlet channel defines a viewport region forming a though hole in the substrate. The first graphene layer overlays the first surface of the substrate, including an interior area of the first channel, the second channel, and the outlet channel. The second graphene layer overlays the first surface of the substrate, including open regions defined by the first channel, the second channel, and the outlet channel.

  7. Detection of Streptococcus agalactiae colonization in pregnant women by using combined swab cultures: cross-sectional prevalence study

    Directory of Open Access Journals (Sweden)

    Camila Marconi

    Full Text Available CONTEXT AND OBJECTIVE: Maternal Streptococcus agalactiae colonization and early-onset neonatal sepsis have aroused interest in the worldwide literature. Streptococcal neonatal disease is associated with significant morbidity and mortality in the perinatal period, especially among premature neonates. The aim of this study was to assess the prevalence of maternal streptococcal colonization by using combined swab cultures, compared with swab collection from a single site. DESIGN AND SETTING: Cross-sectional study at Faculdade de Medicina de Botucatu, Universidade Estadual Paulista. METHODS: Samples were obtained from 405 patients at gestational ages of 35 to 37 weeks. Swabs from the perianal (rectal region, vaginal introitus and upper lateral vaginal vault were cultured in Todd-Hewitt selective broth. Colonies suggestive of Streptococcus agalactiae were subjected to the catalase and CAMP (Christie, Atkins, Munch-Petersen tests. To evaluate the positivity of combined swab cultures, Tukey's test was used for comparison of proportions. RESULTS: The prevalence of streptococcal colonization was 25.4%. Among the patients with positive cultures, 28.1% had this at only one collection site, 24.2% simultaneously at two sites and 47.5% at all three sites. Associating the swabs from two collection sites significantly increased streptococcal isolation, compared with a single swab (P < 0.05, except for perianal (rectal collection. Use of combined swabs from three collection sites showed statistically higher isolation rates. CONCLUSION: In combined swab cultures collected from three collection sites, the prevalence of maternal Streptococcus agalactiae colonization was higher than in swabs collected from a single site.

  8. Occurrence of Tropheryma whipplei during diarrhea in Hajj pilgrims: a PCR analysis of paired rectal swabs.

    Science.gov (United States)

    Gautret, Philippe; Benkouiten, Samir; Parola, Philippe; Brouqui, Philippe; Memish, Ziad; Raoult, Didier

    2014-01-01

    Tropheryma whipplei was recently associated with gastroenteritis in children. We hypothesize that T. whipplei may be a contributing microbe in traveller's diarrhea. The presence of T. whipplei was investigated by PCR on rectal swab samples of Hajj pilgrims before and after travelling to the Kingdom of Saudi Arabia (KSA). Additionally a rectal swab was performed at the time of diarrhea for some pilgrims. A total of 129 pilgrims underwent rectal swab samples before departure and on return. All pilgrims were negative for T. whipplei before travel. One pilgrim (0.8%) was positive on return but did not reported diarrhea. A total of 30 pilgrims (23.3%) experienced diarrhea during the stay in the KSA. Nine pilgrims with diarrhea underwent the additional rectal swab during their diarrhea episode, two of them were positive for T. whipplei. This work suggests that T. whipplei may be associated with adult traveller's diarrhea, by finding T. whipplei DNA individuals negative before and after the episode of diarrhea. Further study addressing this issue in larger cohorts of Hajj pilgrims with systematic sampling at the time of diarrheal episode may help to understand the potential role of T. whipplei in traveller's diarrhea. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Comparison of urine with urethral swabs for the detection of ...

    African Journals Online (AJOL)

    Although cell culture has been regarded as the 'gold standard' for C. ... immunofluorescence as a 'gold standard', because of loss of chlamydial ... Specimens for EIA were collected with. Dacron swabs on aluminium shafts with prescored plastic handles. Swabs were immersed and broken off in 1 ml EIA transport media ...

  10. Molecular Comparison of Bacterial Communities on Peripheral Intravenous Catheters and Matched Skin Swabs.

    Directory of Open Access Journals (Sweden)

    Md Abu Choudhury

    Full Text Available Skin bacteria at peripheral intravenous catheter (PIVC insertion sites pose a serious risk of microbial migration and subsequent colonisation of PIVCs, and the development of catheter related bloodstream infections (CRBSIs. Common skin bacteria are often associated with CRBSIs, therefore the bacterial communities at PIVC skin sites are likely to have major implications for PIVC colonisation. This study aimed to determine the bacterial community structures on skin at PIVC insertion sites and to compare the diversity with associated PIVCs. A total of 10 PIVC skin site swabs and matching PIVC tips were collected by a research nurse from 10 hospitalised medical/surgical patients at catheter removal. All swabs and PIVCs underwent traditional culture and high-throughput sequencing. The bacterial communities on PIVC skin swabs and matching PIVCs were diverse and significantly associated (correlation coefficient = 0.7, p<0.001. Methylobacterium spp. was the dominant genus in all PIVC tip samples, but not so for skin swabs. Sixty-one percent of all reads from the PIVC tips and 36% of all reads from the skin swabs belonged to this genus. Staphylococcus spp., (26%, Pseudomonas spp., (10% and Acinetobacter spp. (10% were detected from skin swabs but not from PIVC tips. Most skin associated bacteria commonly associated with CRBSIs were observed on skin sites, but not on PIVCs. Diverse bacterial communities were observed at skin sites despite skin decolonization at PIVC insertion. The positive association of skin and PIVC tip communities provides further evidence that skin is a major source of PIVC colonisation via bacterial migration but microbes present may be different to those traditionally identified via culture methods. The results provide new insights into the colonisation of catheters and potential pathogenesis of bacteria associated with CRBSI, and may assist in developing new strategies designed to reduce the risk of CRBSI.

  11. Swabbing often fails to detect amphibian Chytridiomycosis under conditions of low infection load.

    Directory of Open Access Journals (Sweden)

    Jaehyub Shin

    Full Text Available The pathogenic chytrid fungus, Batrachochytrium dendrobatidis (denoted Bd, causes large-scale epizootics in naïve amphibian populations. Intervention strategies to rapidly respond to Bd incursions require sensitive and accurate diagnostic methods. Chytridiomycosis usually is assessed by quantitative polymerase chain reaction (qPCR amplification of amphibian skin swabs. Results based on this method, however, sometimes yield inconsistent results on infection status and inaccurate scores of infection intensity. In Asia and other regions where amphibians typically bear low Bd loads, swab results are least reliable. We developed a Bd-sampling method that collects zoospores released by infected subjects into an aquatic medium. Bd DNA is extracted by filters and amplified by nested PCR. Using laboratory colonies and field populations of Bombina orientalis, we compare results with those obtained on the same subjects by qPCR of DNA extracted from swabs. Many subjects, despite being diagnosed as Bd-negative by conventional methods, released Bd zoospores into collection containers and thus must be considered infected. Infection loads determined from filtered water were at least 1000 times higher than those estimated from swabs. Subjects significantly varied in infection load, as they intermittently released zoospores, over a 5-day period. Thus, the method might be used to compare the infectivity of individuals and study the periodicity of zoospore release. Sampling methods based on water filtration can dramatically increase the capacity to accurately diagnose chytridiomycosis and contribute to a better understanding of the interactions between Bd and its hosts.

  12. DNA from buccal swabs suitable for high-throughput SNP multiplex analysis.

    Science.gov (United States)

    McMichael, Gai L; Gibson, Catherine S; O'Callaghan, Michael E; Goldwater, Paul N; Dekker, Gustaaf A; Haan, Eric A; MacLennan, Alastair H

    2009-12-01

    We sought a convenient and reliable method for collection of genetic material that is inexpensive and noninvasive and suitable for self-collection and mailing and a compatible, commercial DNA extraction protocol to meet quantitative and qualitative requirements for high-throughput single nucleotide polymorphism (SNP) multiplex analysis on an automated platform. Buccal swabs were collected from 34 individuals as part of a pilot study to test commercially available buccal swabs and DNA extraction kits. DNA was quantified on a spectrofluorometer with Picogreen dsDNA prior to testing the DNA integrity with predesigned SNP multiplex assays. Based on the pilot study results, the Catch-All swabs and Isohelix buccal DNA isolation kit were selected for our high-throughput application and extended to a further 1140 samples as part of a large cohort study. The average DNA yield in the pilot study (n=34) was 1.94 microg +/- 0.54 with a 94% genotyping pass rate. For the high-throughput application (n=1140), the average DNA yield was 2.44 microg +/- 1.74 with a >or=93% genotyping pass rate. The Catch-All buccal swabs are a convenient and cost-effective alternative to blood sampling. Combined with the Isohelix buccal DNA isolation kit, they provided DNA of sufficient quantity and quality for high-throughput SNP multiplex analysis.

  13. UriSwab: an effective transport medium for nucleic acid detection of Chlamydia trachomatis, Mycoplasma genitalium and Neisseria gonorrhoeae.

    Science.gov (United States)

    Costa, Anna-Maria G; Garland, Suzanne M; Guy, Rebecca; Wand, Handan; Tabrizi, Sepehr N

    2017-11-01

    Background Patient self-sampling allows for remote collection and return to clinic or laboratory by post. Urine samples, although convenient, are challenging to post. This study evaluated UriSwab (Copan, Brescia, Italy) as a collection and transport vessel for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Mycoplasma genitalium (MG) detection by polymerase chain reaction, compared with flocked swab and neat urine. Five replicates of each specimen type were prepared from previously characterised urine samples (n=330), stored at room temperature (RT) or 37°C, then extracted on day 1, 3, 7, 10 and 16 (VERSANT kPCR Sample Prep System, Siemens, Munich, Germany). Crossing thresholds (Cq) from CT and NG detection (VERSANT CT/GC DNA 1.0 assay kit, Siemens) and MG detection (real-time polymerase chain reaction assay) were compared using logistic regression, stratified by sample type, temperature and analyte. Mixed-model statistical techniques were used to assess correlation between repeated observations. UriSwab showed an increasing trend in Cq values at RT and 37°C for CT and NG, and RT for MG (all P<0.01). UriSwab was not statistically significantly different to neat urine, except CT at RT (0.83, 95% confidence interval: 0.51-1.15). Flocked swab similarly showed increasing Cq values at 37°C for CT, a significant decreasing trend at RT for MG and increasing trend at 37°C for MG. Flocked swab was not statistically significantly different from neat urine at RT and 37°C for CT and MG. UriSwab allows transport of urine for CT, NG and MG detection regardless of storage time or temperature, suggesting that CT and NG are stable for up to 16 days and MG up to 10 days.

  14. Pilot study of participant-collected nasal swabs for acute respiratory infections in a low-income, urban population

    Directory of Open Access Journals (Sweden)

    Vargas CY

    2016-01-01

    Full Text Available Celibell Y Vargas,1 Liqun Wang,1 Yaritza Castellanos de Belliard,1 Maria Morban,1 Hilbania Diaz,1 Elaine L Larson,2,3 Philip LaRussa,1 Lisa Saiman,1,4 Melissa S Stockwell1,5,6 1Department of Pediatrics, 2School of Nursing, 3Department of Epidemiology, Mailman School of Public Health, Columbia University, 4Department of Infection Prevention and Control, NewYork-Presbyterian Hospital, 5Department of Population and Family Health, Mailman School of Public Health, Columbia University, 6NewYork-Presbyterian Hospital, New York, NY, USA Objective: To assess the feasibility and validity of unsupervised participant-collected nasal swabs to detect respiratory pathogens in a low-income, urban minority population. Methods: This project was conducted as part of an ongoing community-based surveillance study in New York City to identify viral etiologies of acute respiratory infection. In January 2014, following sample collection by trained research assistants, participants with acute respiratory infection from 30 households subsequently collected and returned a self-collected/parent-collected nasal swab via mail. Self/parental swabs corresponding with positive reverse transcription polymerase chain reaction primary research samples were analyzed. Results: Nearly all (96.8%, n=30/31 households agreed to participate; 100% reported returning the sample and 29 were received (median time: 8 days. Most (18; 62.1% of the primary research samples were positive. For eight influenza-positive research samples, seven (87.5% self-swabs were also positive. For ten other respiratory pathogen-positive research samples, eight (80.0% self-swabs were positive. Sensitivity of self-swabs for any respiratory pathogen was 83.3% and 87.5% for influenza, and specificity for both was 100%. There was no relationship between level of education and concordance of results between positive research samples and their matching participant swab. Conclusion: In this pilot study, self-swabbing

  15. Evaluation of Three Swabbing Devices for Detection of Listeria monocytogenes on Different Types of Food Contact Surfaces

    Directory of Open Access Journals (Sweden)

    Evy Lahou

    2014-01-01

    Full Text Available Listeria monocytogenes can adhere to different types of food contact surfaces within a food processing environment. Therefore, environmental sampling devices should be capable of detecting unacceptable contamination. In this study, a sponge-stick, foam spatula and an environmental swab were evaluated on their ability to detect low concentrations of L. monocytogenes on different types of food contact surfaces. A cocktail of four L. monocytogenes serotypes was inoculated with a concentration of 100 CFU/250 cm2 onto stainless steel (SS, high density polyethylene (HDPE and rubber surfaces in a 250 cm2 area. Immediately after inoculation and after 1 h exposure, the surfaces were swabbed with the different swabbing devices. The results of the study show only minor differences in the ability of the swabbing devices to detect L. monocytogenes. All devices were capable to detect the contamination immediately after inoculation. However, when the surfaces were allowed to air-dry for 1 h, L. monocytogenes was undetected in 11.1% of the samples (n = 27 with the sponge stick, in 7.4% of the samples (n = 27 with the foam spatula and in 3.7% of the samples (n = 27 with the environmental swab, especially on SS surfaces. The detection ability of the different devices for L. monocytogenes can be concluded to be rather high on different types of food contact surfaces.

  16. Imperfect pathogen detection from non-invasive skin swabs biases disease inference

    Science.gov (United States)

    DiRenzo, Graziella V.; Grant, Evan H. Campbell; Longo, Ana; Che-Castaldo, Christian; Zamudio, Kelly R.; Lips, Karen

    2018-01-01

    1. Conservation managers rely on accurate estimates of disease parameters, such as pathogen prevalence and infection intensity, to assess disease status of a host population. However, these disease metrics may be biased if low-level infection intensities are missed by sampling methods or laboratory diagnostic tests. These false negatives underestimate pathogen prevalence and overestimate mean infection intensity of infected individuals. 2. Our objectives were two-fold. First, we quantified false negative error rates of Batrachochytrium dendrobatidis on non-invasive skin swabs collected from an amphibian community in El Copé, Panama. We swabbed amphibians twice in sequence, and we used a recently developed hierarchical Bayesian estimator to assess disease status of the population. Second, we developed a novel hierarchical Bayesian model to simultaneously account for imperfect pathogen detection from field sampling and laboratory diagnostic testing. We evaluated the performance of the model using simulations and varying sampling design to quantify the magnitude of bias in estimates of pathogen prevalence and infection intensity. 3. We show that Bd detection probability from skin swabs was related to host infection intensity, where Bd infections caused by skin swabs in persisting host communities with low-level infections. More generally, our results have implications for study designs in other disease systems, particularly those with similar objectives, biology, and sampling decisions. 4. Uncertainty in pathogen detection is an inherent property of most sampling protocols and diagnostic tests, where the magnitude of bias depends on the study system, type of infection, and false negative error rates. Given that it may be difficult to know this information in advance, we advocate that the most cautious approach is to assume all errors are possible and to accommodate them by adjusting sampling designs. The modeling framework presented here improves the accuracy in

  17. Unlocking the story in the swab: A new genotyping assay for the amphibian chytrid fungus Batrachochytrium dendrobatidis.

    Science.gov (United States)

    Byrne, Allison Q; Rothstein, Andrew P; Poorten, Thomas J; Erens, Jesse; Settles, Matthew L; Rosenblum, Erica Bree

    2017-11-01

    One of the most devastating emerging pathogens of wildlife is the chytrid fungus, Batrachochytrium dendrobatidis (Bd), which affects hundreds of amphibian species around the world. Genomic data from pure Bd cultures have advanced our understanding of Bd phylogenetics, genomic architecture and mechanisms of virulence. However, pure cultures are laborious to obtain and whole-genome sequencing is comparatively expensive, so relatively few isolates have been genetically characterized. Thus, we still know little about the genetic diversity of Bd in natural systems. The most common noninvasive method of sampling Bd from natural populations is to swab amphibian skin. Hundreds of thousands of swabs have been collected from amphibians around the world, but Bd DNA collected via swabs is often low in quality and/or quantity. In this study, we developed a custom Bd genotyping assay using the Fluidigm Access Array platform to amplify 192 carefully selected regions of the Bd genome. We obtained robust sequence data for pure Bd cultures and field-collected skin swabs. This new assay has the power to accurately discriminate among the major Bd clades, recovering the basic tree topology previously revealed using whole-genome data. Additionally, we established a critical value for initial Bd load for swab samples (150 Bd genomic equivalents) above which our assay performs well. By leveraging advances in microfluidic multiplex PCR technology and the globally distributed resource of amphibian swab samples, noninvasive skin swabs can now be used to address critical spatial and temporal questions about Bd and its effects on declining amphibian populations. © 2017 John Wiley & Sons Ltd.

  18. Gossypiboma – the retained surgical swab: An enduring clinical ...

    African Journals Online (AJOL)

    that raise clinical suspicion, of a condition that may be elusive in presentation on ... A review of the English literature reporting retained abdominal swabs between 1992 and 2012 revealed 100 cases. .... help to explain the variety of associated.

  19. Validation of a same-day real-time PCR method for screening of meat and carcass swabs for Salmonella

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Krause, Michael; Josefsen, Mathilde Hartmann

    2009-01-01

    swabs). The relative accuracy was 99%, relative detection level 100%, relative sensitivity 103% and relative specificity 100%. The collaborative trial included six laboratories testing minced meat, poultry neck-skins, and carcass swabs as un-inoculated samples and samples artificially contaminated....... Partly based on results obtained in this study, the method has obtained NordVal approval for analysis of Salmonella in meat and carcass swabs. The PCR method was transferred to a production laboratory and the performance was compared with the BAX Salmonella test on 39 pork samples artificially......Background: One of the major sources of human Salmonella infections is meat. Therefore, efficient and rapid monitoring of Salmonella in the meat production chain is necessary. Validation of alternative methods is needed to prove that the performance is equal to established methods. Very few...

  20. Abdominal fibrosarcoma associated with a retained surgical swab in a dog.

    Science.gov (United States)

    Rayner, E L; Scudamore, C L; Francis, I; Schöniger, S

    2010-07-01

    An abdominal fibrosarcoma surrounding a retained surgical swab was identified in a 3-year-old neutered female rottweiler dog presented with chronic inappetence and lethargy. Laparotomy revealed a mass within the omentum, multiple hepatic masses and enlarged mesenteric lymph nodes. The dog was humanely destroyed and submitted for necropsy examination. Microscopically, the omental mass was consistent with a sarcoma surrounding centrally located fibres of foreign material and was infiltrated by epithelioid macrophages containing intracytoplasmic fibre fragments. Sarcoma tissue was also present in mesenteric lymph nodes, liver, spleen and lungs, and some affected lymph nodes contained intralesional epithelioid macrophages with fibre fragments. Immunohistochemical and electron microscopical examinations were consistent with a diagnosis of fibrosarcoma. By fibre analysis and electron microscopy, the intratumoural fibres were identified as cotton fibres with features identical to those obtained from a surgical swab. To our knowledge this is the first description of an abdominal fibrosarcoma associated with a retained surgical swab in a dog. Other examples of foreign body-associated sarcomas in the veterinary literature are vaccine- and implant-induced sarcomas. (c) 2010 Elsevier Ltd. All rights reserved.

  1. Estimating hadron contamination of electron samples in Pb-Pb collisions at low momenta using ALICE

    Energy Technology Data Exchange (ETDEWEB)

    Brass, Martin [Physikalisches Institut, Universitaet Heidelberg (Germany); Collaboration: ALICE-Collaboration

    2015-07-01

    A Large Ion Collider Experiment at the LHC is exploring a new state of matter at high energy densities in Pb-Pb collisions. Electrons from heavy-flavor decays are interesting probes of the properties of this state, since charm and beauty quarks - produced in initial hard scatterings - experience the whole evolution of the state. Due to high multiplicities in Pb-Pb collisions and to the limited separation power of the ALICE Time-Of-Flight (TOF) detector between electrons and pions at momenta above 1 GeV/c, there is a large hadronic background in the observed electron samples. Furthermore a significant amount of protons, kaons and deuterons are misidentified by TOF and contaminate the electron sample, too. Many Heavy-Flavor-Electron analyses rely on a precise estimation of the contamination. The presented analysis uses measured energy loss distributions of the ALICE Time Projection Chamber as templates for a least squares fit routine to estimate the amount of hadronic background in the electron sample at momenta above 0.4 GeV/c. To describe the energy loss distributions of the misidentified particles, the measured distributions of particles which are identified by TOF are used. The energy loss distribution for pions is obtained using the ALICE Transition Radiation Detector which provides good separation between pions and electrons.

  2. Measurement of actinides in environmental samples by Photo-Electron Rejecting Alpha Liquid Scintillation

    Energy Technology Data Exchange (ETDEWEB)

    Cadieux, J.R. [Westinghouse Savannah River Co., Aiken, SC (United States); Clark, S. [Savannah River Ecology Lab., Univ. of Georgia (United States); Fjeld, R.A.; Reboul, S.; Sowder, A. [Clemson Univ., SC (United States). Dept. of Environmental Systems Engineering

    1994-05-01

    This work describes the adaptation of extractive scintillation with a Photo-Electron Rejecting Alpha Liquid Scintillation (PERALS) (ORDELA, Inc.) spectrometer to the analysis of actinides in environmental samples from the Savannah River Site (SRS). Environmental quality assurance standards and actual water samples were treated by one of two methods; either a two step direct extraction, or for more complex samples, pretreatment by an extraction chromatographic separation prior to measurement of the alpha activity by PERALS.

  3. Detection of strep throat causing bacterium directly from medical swabs by touch spray - mass spectrometry

    OpenAIRE

    Jarmusch, Alan K.; Pirro, Valentina; Kerian, Kevin S.; Cooks, Graham

    2014-01-01

    Strep throat causing Streptococcus pyogenes was detected in vitro and in simulated clinical samples by performing touch spray ionization - mass spectrometry. MS analysis took only seconds to reveal characteristic bacterial and human lipids. Medical swabs were used as the substrate for ambient ionization. This work constitutes the initial step in developing a noninvasive MS-based test for clinical diagnosis of strep throat. It is limited to the single species, S. pyogenes, which is responsible...

  4. Measurements of NaI(Tl) electron response: comparison of different samples

    Energy Technology Data Exchange (ETDEWEB)

    Hull, Giulia; Choong, Woon-Seng; Moses, William W.; Bizarri, Gregory; Valentine, John D.; Payne, Stephen A.; Cherepy, Nerine J.; Reutter, Bryan W.

    2008-12-10

    This paper measures the sample to sample variation in the light yield proportionality of NaI(Tl), and so explores whether this is an invariant characteristic of the material or whether it depends on the chemical and physical properties of the tested samples. We report on the electron response of nine crystals of NaI(Tl), differing in shape, volume, age, manufacturer and quality. The proportionality has been measured at the SLYNCI facility in the energy range between 3.5 to 460 keV. We observe that while samples produced by the same manufacturer at approximately the same time have virtually identical electron response curves, there are significant sample to sample variations among crystals produced by different manufacturers or at different times. In an effort to correlate changes in the electron response with details of the scintillation mechanism, we characterized other scintillation properties, including the gamma response and the x-ray excited emission spectra and decay times, for the nine crystals. While sample to sample differences in these crystals were observed, we have been unable to identify the underlying fundamental mechanisms that are responsible for these differences.

  5. Ninhydrin treatment as a screening method for the suitability of swabs taken from contact stains for DNA analysis.

    Science.gov (United States)

    Anslinger, K; Selbertinger, U; Bayer, B; Rolf, B; Eisenmenger, W

    2004-04-01

    More and more swabs containing unknown traces of biological material are submitted for forensic DNA analysis. Most of the samples are swabs taken from handled items such as tools, weapons and handles etc. Therefore, we tried to develop a screening method in order to focus the investigation on samples containing biomolecules, such as amino acids which might be associated with nucleic acids. A total of 285 swabs taken from various items collected during crime scene investigations were treated with ninhydrin which leads to a purple colour for samples containing amino acids. Of the swabs 158 were classified as ninhydrin positive and 76% of these samples yielded DNA profiles that fulfil the criteria for inclusion in the German national DNA database (profile frequency greater than 1 in 100,000) or in DNA mixtures which could at least be compared with suspects. In comparison only 9% of the 127 samples shown to be ninhydrin negative, revealed a usable DNA profile. Consequently, ninhydrin treatment was found to be an effective screening method which resulted in an increase in the rate of successfully typed samples and subsequently in a reduction of the costs due to the lower number of samples that needed to be typed.

  6. A typical proficiency testing programmes sample design for electrical and electronic product

    Science.gov (United States)

    Wang, T. T.; Zhang, H.; Xie, L. L.; Wang, Y. Y.

    2017-04-01

    Creepage distance and clearance testing are the basic testing items in the safety standards for almost all electrical and electronic products. A typical sample group is designed in this paper for the purpose of proficiency testing programmes. The sample group is composed of two kinds of circuit board. The length of the creepage distance of the two circuit boards in pollution degree 2 and 3 are the same but with different paths. This sample group includes three testing points. This sample group is designed beneficial for numerical statistics and avoiding the data complicity in the laboratory. It can be used for effective laboratory monitoring.

  7. Reduced electron exposure for energy-dispersive spectroscopy using dynamic sampling

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yan; Godaliyadda, G. M. Dilshan; Ferrier, Nicola; Gulsoy, Emine B.; Bouman, Charles A.; Phatak, Charudatta

    2018-01-01

    Analytical electron microscopy and spectroscopy of biological specimens, polymers, and other beam sensitive materials has been a challenging area due to irradiation damage. There is a pressing need to develop novel imaging and spectroscopic imaging methods that will minimize such sample damage as well as reduce the data acquisition time. The latter is useful for high-throughput analysis of materials structure and chemistry. In this work, we present a novel machine learning based method for dynamic sparse sampling of EDS data using a scanning electron microscope. Our method, based on the supervised learning approach for dynamic sampling algorithm and neural networks based classification of EDS data, allows a dramatic reduction in the total sampling of up to 90%, while maintaining the fidelity of the reconstructed elemental maps and spectroscopic data. We believe this approach will enable imaging and elemental mapping of materials that would otherwise be inaccessible to these analysis techniques.

  8. PCR-based assay to detect sheeppox virus in ocular, nasal, and rectal swabs from infected Moroccan sheep.

    Science.gov (United States)

    Zro, K; Azelmat, S; Bendouro, Y; Kuhn, J H; El Fahime, E; Ennaji, M M

    2014-08-01

    Sheeppox is now enzootic in Morocco. The development of a reliable method for rapid diagnosis of the disease is a central part of any control strategy. The aim of this study is to determine the diagnostic value of a variety of clinical samples such as ovine nasal, ocular or rectal swabs for the detection of sheeppox virus (SPPV) by qualitative conventional polymerase chain reaction (PCR), using a single pair of primers targeting the inverted terminal repeats of the SPPV InS-1 strain, a virulent field isolate. Swab and blood samples were collected from forty animals naturally infected with SPPV who had clinical signs of sheeppox. All animals tested PCR-positive for SPPV. Positive results were obtained infrequently with blood samples, whereas swab samples from at least two sites (nasal, ocular, rectal) were positive per evaluated animal. These results indicate that swab samples are suitable for quantitative molecular SPPV diagnosis. PCR product sequences obtained from all types of sheep samples proved to be identical to the corresponding regions of sheeppox virus strain Romania 65. Copyright © 2014. Published by Elsevier B.V.

  9. A Transmission Electron Microscope Investigation of Space Weathering Effects in Hayabusa Samples

    Science.gov (United States)

    Keller, Lindsay P.; Berger, Eve L.

    2014-01-01

    The Hayabusa mission to asteroid 25143 Itokawa successfully returned the first direct samples of the regolith from the surface of an asteroid. The Hayabusa samples thus present a special opportunity to directly investigate the evolution of asteroidal surfaces, from the development of the regolith to the study of the more complex effects of space weathering. Here we describe the mineralogy, microstructure and composition of three Hayabusa mission particles using transmission electron microscope (TEM) techniques

  10. GraFix: sample preparation for single-particle electron cryomicroscopy.

    Science.gov (United States)

    Kastner, Berthold; Fischer, Niels; Golas, Monika Mariola; Sander, Bjoern; Dube, Prakash; Boehringer, Daniel; Hartmuth, Klaus; Deckert, Jochen; Hauer, Florian; Wolf, Elmar; Uchtenhagen, Hannes; Urlaub, Henning; Herzog, Franz; Peters, Jan Michael; Poerschke, Dietmar; Lührmann, Reinhard; Stark, Holger

    2008-01-01

    We developed a method, named GraFix, that considerably improves sample quality for structure determination by single-particle electron cryomicroscopy (cryo-EM). GraFix uses a glycerol gradient centrifugation step in which the complexes are centrifuged into an increasing concentration of a chemical fixation reagent to prevent aggregation and to stabilize individual macromolecules. The method can be used to prepare samples for negative-stain, cryo-negative-stain and, particularly, unstained cryo-EM.

  11. Identification of rhabdoviral sequences in oropharyngeal swabs from German and Danish bats

    DEFF Research Database (Denmark)

    Fischer, Melina; Freuling, Conrad M.; Müller, Thomas

    2014-01-01

    Background: In the frame of active lyssavirus surveillance in bats, oropharyngeal swabs from German (N = 2297) and Danish (N = 134) insectivorous bats were investigated using a newly developed generic pan-lyssavirus real-time reverse transcriptase PCR (RT-qPCR).Findings: In total, 15 RT......-qPCR positive swabs were detected. Remarkably, sequencing of positive samples did not confirm the presence of bat associated lyssaviruses but revealed nine distinct novel rhabdovirus-related sequences. Conclusions: Several novel rhabdovirus-related sequences were detected both in German and Danish insectivorous...... bats. The results also prove that the novel generic pan-lyssavirus RT-qPCR offers a very broad detection range that allows the collection of further valuable data concerning the broad and complex diversity within the family Rhabdoviridae....

  12. Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography.

    Science.gov (United States)

    Trépout, Sylvain; Bastin, Philippe; Marco, Sergio

    2017-03-12

    This report describes a protocol for preparing thick biological specimens for further observation using a scanning transmission electron microscope. It also describes an imaging method for studying the 3D structure of thick biological specimens by scanning transmission electron tomography. The sample preparation protocol is based on conventional methods in which the sample is fixed using chemical agents, treated with a heavy atom salt contrasting agent, dehydrated in a series of ethanol baths, and embedded in resin. The specific imaging conditions for observing thick samples by scanning transmission electron microscopy are then described. Sections of the sample are observed using a through-focus method involving the collection of several images at various focal planes. This enables the recovery of in-focus information at various heights throughout the sample. This particular collection pattern is performed at each tilt angle during tomography data collection. A single image is then generated, merging the in-focus information from all the different focal planes. A classic tilt-series dataset is then generated. The advantage of the method is that the tilt-series alignment and reconstruction can be performed using standard tools. The collection of through-focal images allows the reconstruction of a 3D volume that contains all of the structural details of the sample in focus.

  13. Proton Transmitting Energy Spectra and Transmission Electron Microscope Examinations of Biological Samples

    Science.gov (United States)

    Tan, Chun-yu; Xia, Yue-yuan; Zhang, Jian-hua; Mu, Yu-guang; Wang, Rui-jin; Liu, Ji-tian; Liu, Xiang-dong; Yu, Zeng-liang

    1999-02-01

    Transmission energy spectra of 530 keV H+ ion penetrating 140 μm thick seed coat of maize and fruit peel of grape with thickness of 100 μm were measured. The result indicates that these thick biological targets, as seen by the penetrating ions, are inhomogeneous, and there are open "channel like" paths along which the incident ions can transmit the targets easily. While most of the incident ions are stopped in the targets, some of the transmitting ions only lose a small fraction of their initial incident energy. The transmission energy spectra show a pure electronic stopping feature. Transmission electron microscope (TEM) micrographes taken from the samples of seed coat of maize and fruit peel of tomato with thickness of 60 μm indicate that 150 keV electron beam from the TEM can penetrate the thick samples to give very good images with clear contrasts.

  14. Rapid combined light and electron microscopy on large frozen biological samples

    NARCIS (Netherlands)

    Vogels, I. M. C.; Hoeben, K. A.; van Noorden, C. J. F.

    2009-01-01

    P>The use of large unfixed frozen tissue samples (10 x 10 x 5 mm(3)) for combined light microscopy (LM) and electron microscopy (EM) is described. First, cryostat sections are applied for various LM histochemical approaches including in situ hybridization, immunohistochemistry and metabolic mapping

  15. Determination of selenium in biological samples by gas-liquid chromatography with electron-capture detection.

    Science.gov (United States)

    Poole, C F; Evans, N J; Wibberley, D G

    1977-06-01

    Selenium can be determined quantitatively in biological samples after nitric acid-magnesium nitrate digestion and formation of 5-nitropiazselenole, by extraction into toluene for gas-liquid chromatography with electron-capture detection. The method is suitable for the determination of selenium in orchard leaves, bovine liver and human placenta, hair, blood and urine.

  16. Evaluation of an autoclave resistant anatomic nose model for the testing of nasal swabs.

    Science.gov (United States)

    Bartolitius, Lennart; Frickmann, Hagen; Warnke, Philipp; Ottl, Peter; Podbielski, Andreas

    2014-09-01

    A nose model that allows for the comparison of different modes of sample acquisition as well as of nasal swab systems concerning their suitability to detect defined quantities of intranasal microorganisms, and further for training procedures of medical staff, was evaluated. Based on an imprint of a human nose, a model made of a silicone elastomer was formed. Autoclave stability was assessed. Using an inoculation suspension containing Staphylococcus aureus and Staphylococcus epidermidis, the model was compared with standardized glass plate inoculations. Effects of inoculation time, mode of sampling, and sample storage time were assessed. The model was stable to 20 autoclaving cycles. There were no differences regarding the optimum coverage from the nose and from glass plates. Optimum sampling time was 1 h after inoculation. Storage time after sampling was of minor relevance for the recovery. Rotating the swab around its own axis while circling the nasal cavity resulted in best sampling results. The suitability of the assessed nose model for the comparison of sampling strategies and systems was confirmed. Without disadvantages in comparison with sampling from standardized glass plates, the model allows for the assessment of a correct sampling technique due to its anatomically correct shape.

  17. Comparison of Easy-Flow Copan Liquid Stuart's and Starplex Swab transport systems for recovery of fastidious aerobic bacteria.

    Science.gov (United States)

    Drake, Cheryl; Barenfanger, Joan; Lawhorn, Jerry; Verhulst, Steven

    2005-03-01

    Because samples are frequently submitted on swabs from distant sites, viability of the organism must be maintained. We compared two transport systems, a new Copan Liquid Stuart's swab with an Easy-Flow swab applicator and the Starplex Liquid Stuart's swab. The purpose of the study was to assess the release and/or recovery of organisms from the Copan system compared to that from Starplex. Triplicate swabs were seeded with 3 dilutions of Neisseria gonorrhoeae, Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae. Although the amount of the initial inoculum was the same for both transport systems, recovery by the roll-plate method at time zero was consistently increased with the Copan system (31 to 87% higher). This is the most important finding in this study. With N. gonorrhoeae, subsequent recoveries were similar for Copan and Starplex but poor for both systems. With N. meningitidis and Haemophilus, higher levels of recovery were clearly obtained with Copan (P < 0.05 to P < 0.001). With Streptococcus, subsequent recoveries for Copan and Starplex were mixed. In conclusion, Copan generally demonstrated better recovery of organisms compared to Starplex even (and especially) at time zero.

  18. Influence of surface topography on the secondary electron yield of clean copper samples.

    Science.gov (United States)

    Hu, Xiao-Chuan; Cao, Meng; Cui, Wan-Zhao

    2016-11-01

    Secondary electron yield (SEY) due to electron impact depends strongly on surface topography. The SEY of copper samples after Ar-ion bombardment is measured in situ in a multifunctional ultrahigh vacuum system. Increasing the ion energy or duration of ion bombardment can even enlarge the SEY, though it is relatively low under moderate bombardment intensity. The results obtained with scanning electron microscopy and atomic force microscopy images demonstrate that many valley structures of original sample surfaces can be smoothed due to ion bombardment, but more hill structures are generated with stronger bombardment intensity. With increasing the surface roughness in the observed range, the maximum SEY decreases from 1.2 to 1.07 at a surface characterized by valleys, while it again increases to 1.33 at a surface spread with hills. This phenomenon indicates that hill and valley structures are respectively effective in increasing and decreasing the SEY. These obtained results thus provide a comprehensive insight into the surface topography influence on the secondary electron emission characteristics in scanning electron microscopy. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Acetic Acid Detection Threshold in Synthetic Wine Samples of a Portable Electronic Nose

    Directory of Open Access Journals (Sweden)

    Miguel Macías Macías

    2012-12-01

    Full Text Available Wine quality is related to its intrinsic visual, taste, or aroma characteristics and is reflected in the price paid for that wine. One of the most important wine faults is the excessive concentration of acetic acid which can cause a wine to take on vinegar aromas and reduce its varietal character. Thereby it is very important for the wine industry to have methods, like electronic noses, for real-time monitoring the excessive concentration of acetic acid in wines. However, aroma characterization of alcoholic beverages with sensor array electronic noses is a difficult challenge due to the masking effect of ethanol. In this work, in order to detect the presence of acetic acid in synthetic wine samples (aqueous ethanol solution at 10% v/v we use a detection unit which consists of a commercial electronic nose and a HSS32 auto sampler, in combination with a neural network classifier (MLP. To find the characteristic vector representative of the sample that we want to classify, first we select the sensors, and the section of the sensors response curves, where the probability of detecting the presence of acetic acid will be higher, and then we apply Principal Component Analysis (PCA such that each sensor response curve is represented by the coefficients of its first principal components. Results show that the PEN3 electronic nose is able to detect and discriminate wine samples doped with acetic acid in concentrations equal or greater than 2 g/L.

  20. Assessment of the contribution of electron microscopy to nanoparticle characterization sampled with two cascade impactors.

    Science.gov (United States)

    Noël, Alexandra; L'Espérance, Gilles; Cloutier, Yves; Plamondon, Philippe; Boucher, Julie; Philippe, Suzanne; Dion, Chantal; Truchon, Ginette; Zayed, Joseph

    2013-01-01

    This study assessed the contribution of electron microscopy to the characterization of nanoparticles and compared the degree of variability in sizes observed within each stage when sampled by two cascade impactors: an Electrical Low Pressure Impactor (ELPI) and a Micro-Orifice Uniform Deposit Impactor (MOUDI). A TiO(2) nanoparticle (5 nm) suspension was aerosolized in an inhalation chamber. Nanoparticles sampled by the impactors were collected on aluminum substrates or TEM carbon-coated copper grids using templates, specifically designed in our laboratories, for scanning and transmission electron microscopy (SEM, TEM) analysis, respectively. Nanoparticles were characterized using both SEM and TEM. Three different types of diameters (inner, outer, and circular) were measured by image analysis based on count and volume, for each impactor stage. Electron microscopy, especially TEM, is well suited for the characterization of nanoparticles. The MOUDI, probably because of the rotation of its collection stages, which can minimize the resuspension of particles, gave more stable results and smaller geometric standard deviations per stage. Our data suggest that the best approach to estimate particle size by electron microscopy would rely on geometric means of measured circular diameters. Overall, the most reliable data were provided by the MOUDI and the TEM sampling technique on carbon-coated copper grids for this specific experiment. This study indicates interesting findings related to the assessment of impactors combined with electron microscopy for nanoparticle characterization. For future research, since cascade impactors are extensively used to characterize nano-aerosol exposure scenarios, high-performance field emission scanning electron microscopy (FESEM) should also be considered.

  1. Accuracy of self-collected vaginal dry swabs using the Xpert human papillomavirus assay.

    Directory of Open Access Journals (Sweden)

    Rosa Catarino

    Full Text Available Polymerase chain reaction-based Xpert human papillomavirus (HPV assay is a rapid test that detects high-risk HPV (hrHPV infection. This point-of-care test is usually performed by collecting a cervical specimen in a vial of PreservCyt® transport medium. We compared HPV test positivity and accuracy between self-collected sample with a dry swab (s-DRY versus physician-collected cervical sampling using a broom like brush and immediate immersion in PreservCyt (dr-WET.In this cross-sectional study, we recruited 150 women ≥ 18 years old attending the colposcopy clinic in the University Hospital of Geneva. Each participant first self-collected a vaginal sample using a dry swab and then the physician collected a cervical specimen in PreservCyt. HPV analysis was performed with Xpert. Part of the PreservCyt-collected sample was used for hrHPV detection with the cobas® HPV test. HPV test positivity and performance of the two collection methods was compared.HPV positivity was 49.1% for s-DRY, 41.8% for dr-WET and 46.2% for cobas. Good agreement was found between s-DRY and dr-WET samples (kappa±Standard error (SE = 0.64±0.09,, particularly for low-grade squamous intraepithelial lesions (LSIL+ (kappa±SE = 0.80±0.17. Excellent agreement was found between the two samples for HPV16 detection in general (kappa±SE = 0.91±0.09 and among LSIL+ lesions (kappa±SE = 1.00±0.17. Sensitivities and specificities were, respectively, 84.2% and 47.1%(s-DRY, 73.1% and 58.7%. (dr-WET and 77.8% and 45.7% (cobas for CIN2+ detection. The median delay between sampling and HPV analysis was 7 days for the Xpert HPV assay and 19 days for cobas. There were 36 (24.0% invalid results among s-DRY samples and 4 (2.7% among dr-WET (p = 0.001. Invalid results happened due to the long interval between collection and analysis.Self-collected vaginal dry swabs are a valid alternative to collecting cervical samples in PreservCyt solution for HPV testing with the Xpert HPV assay

  2. Value of bacterial culture of vaginal swabs in diagnosis of vaginal infections

    Directory of Open Access Journals (Sweden)

    Nenadić Dane

    2015-01-01

    Full Text Available Bacground/Aim. Vaginal and cervical swab culture is still very common procedure in our country’s everyday practice whereas simple and rapid diagnostic methods have been very rarely used. The aim of this study was to show that the employment of simple and rapid diagnostic tools [vaginal fluid wet mount microscopy (VFWMM, vaginal pH and potassium hydroxide (KOH test] offers better assessment of vaginal environment than standard microbiologic culture commonly used in Serbia. Methods. This prospective study included 505 asymptomatic pregnant women undergoing VFWMM, test with 10% KOH, determination of vaginal pH and standard culture of cervicovaginal swabs. Combining findings from the procedures was used to make diagnoses of bacterial vaginosis (BV and vaginitis. In addition, the number of polymorphonuclear leukocytes (PMN was determined in each sample and analyzed along with other findings. Infections with Candida albicans and Trichomonas vaginalis were confirmed or excluded by microscopic examination. Results. In 36 (6% patients cervicovaginal swab cultures retrieved several aerobes and facultative anaerobes, whereas in 52 (11% women Candida albicans was isolated. Based on VFWMM findings and clinical criteria 96 (19% women had BV, 19 (4% vaginitis, and 72 (14% candidiasis. Of 115 women with BV and vaginitis, pH 4.5 was found in 5, and of 390 with normal findings 83 (21% had vaginal pH 4.5. Elevated numbers of PMN were found in 154 (30% women - in 83 (54% of them VFWMM was normal. Specificity and sensitivity of KOH test and vaginal pH determination in defining pathological vaginal flora were 95% and 81%, and 79% and 91%, respectively. Conclusion. Cervicovaginal swab culture is expensive but almost non-informative test in clinical practice. The use of simpler and rapid methods as vaginal fluid wet mount microscopy, KOH test and vaginal pH offers better results in diagnosis, and probably in the treatment and prevention of sequels of vaginal

  3. Nasal swab real-time PCR is not suitable for in vivo diagnosis of bovine tuberculosis

    Directory of Open Access Journals (Sweden)

    Fabiana Q. Mayer

    Full Text Available ABSTRACT: Bovine tuberculosis (bTB is a zoonosis causing economic losses and public health risks in many countries. The disease diagnosis in live animals is performed by intradermal tuberculin test, which is based on delayed hypersensitivity reactions. As tuberculosis has complex immune response, this test has limitations in sensitivity and specificity. This study sought to test an alternative approach for in vivo diagnosis of bovine tuberculosis, based on real-time polymerase chain reaction (PCR. DNA samples, extracted from nasal swabs of live cows, were used for SYBR® Green real-time PCR, which is able to differentiate between Mycobacterium tuberculosis and Mycobacterium avium complexes. Statistical analysis was performed to compare the results of tuberculin test, the in vivo gold standard bTB diagnosis method, with real-time PCR, thereby determining the specificity and sensitivity of molecular method. Cervical comparative test (CCT was performed in 238 animals, of which 193 had suitable DNA from nasal swabs for molecular analysis, as indicated by amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH gene, and were included in the study. In total, 25 (10.5% of the animals were CCT reactive, of which none was positive in the molecular test. Of the 168 CCT negative animals, four were positive for M. tuberculosis complex at real time PCR from nasal swabs. The comparison of these results generated values of sensitivity and specificity of 0% and 97.6%, respectively; moreover, low coefficients of agreement and correlation (-0.029 and -0.049, respectively between the results obtained with both tests were also observed. This study showed that real-time PCR from nasal swabs is not suitable for in vivo diagnosis of bovine tuberculosis; thus tuberculin skin test is still the best option for this purpose.

  4. Sample preparation for precise and quantitative electron holographic analysis of semiconductor devices.

    Science.gov (United States)

    Han, Myung-Geun; Li, Jing; Xie, Qianghua; Fejes, Peter; Conner, James; Taylor, Bill; McCartney, Martha R

    2006-08-01

    Wedge polishing was used to prepare one-dimensional Si n-p junction and Si p-channel metal-oxide-silicon field effect transistor (pMOSFET) samples for precise and quantitative electrostatic potential analysis using off-axis electron holography. To avoid artifacts associated with ion milling, cloth polishing with 0.02-microm colloidal silica suspension was used for final thinning. Uniform thickness and no significant charging were observed by electron holography analysis for samples prepared entirely by this method. The effect of sample thickness was investigated and the minimum thickness for reliable results was found to be approximately 160 nm. Below this thickness, measured phase changes were smaller than expected. For the pMOSFET sample, quantitative analysis of two-dimensional electrostatic potential distribution showed that the metallurgical gate length (separation between two extension junctions) was approximately 54 nm, whereas the actual gate length was measured to be approximately 70 nm by conventional transmission electron microscopy. Thus, source and drain junction encroachment under the gate was 16 nm.

  5. Bacteriological pattern of wound swab isolates in patients with ...

    African Journals Online (AJOL)

    PURPOSE: To determine the pattern of bacterial pathogens and their antibiotic sensitivity profile in patients with infected chronic leg ulceration. METHODS: Sixty swab specimens obtained from chronic leg ulcer (CLU) patients were cultured aerobically and the antibiotic sensitivity pattern of the recovered organisms ...

  6. Gossypiboma – the retained surgical swab: An enduring clinical ...

    African Journals Online (AJOL)

    language publications were excluded and abstracts of relevant articles were evaluated for .... [5] The diagnosis of a gossypiboma may be made easily on plain film if the radio-opaque marker is intact; however, ... radio-opaque markers are helpful in identification of retained swabs using plain X-ray. An X-ray is taken on table ...

  7. Bacterial isolates from swab specimens and their susceptibilities to ...

    African Journals Online (AJOL)

    Ciprofloxacin and ofloxacin showed uniform activities against Proteus spp, which showed partial resistance to all agents except sparfloxacin. Multi-drugs resistances are high with all organisms. Many pathogens cause infections in swab sites. The knowledge of causative organisms and their sensitivities are important since ...

  8. Ultrasensitive detection of oncogenic human papillomavirus in oropharyngeal tissue swabs.

    Science.gov (United States)

    Isaac, Andre; Kostiuk, Morris; Zhang, Han; Lindsay, Cameron; Makki, Fawaz; O'Connell, Daniel A; Harris, Jeffrey R; Cote, David W J; Seikaly, Hadi; Biron, Vincent L

    2017-01-14

    The incidence of oropharyngeal squamous cell carcinoma (OPSCC) caused by oncogenic human papillomavirus (HPV) is rising worldwide. HPV-OPSCC is commonly diagnosed by RT-qPCR of HPV E6 and E7 oncoproteins or by p16 immunohistochemistry (IHC). Droplet digital PCR (ddPCR) has been recently reported as an ultra-sensitive and highly precise method of nucleic acid quantification for biomarker analysis. To validate the use of a minimally invasive assay for detection of oncogenic HPV based on oropharyngeal swabs using ddPCR. Secondary objectives were to compare the accuracy of ddPCR swabs to fresh tissue p16 IHC and RT-qPCR, and to compare the cost of ddPCR with p16 IHC. We prospectively included patients with p16+ oral cavity/oropharyngeal cancer (OC/OPSCC), and two control groups: p16- OC/OPSCC patients, and healthy controls undergoing tonsillectomy. All underwent an oropharyngeal swab with ddPCR for quantitative detection of E6 and E7 mRNA. Surgical specimens had p16 IHC performed. Agreement between ddPCR and p16 IHC was determined for patients with p16 positive and negative OC/OPSCC as well as for healthy control patients. The sensitivity and specificity of ddPCR of oropharyngeal swabs were calculated against p16 IHC for OPSCC. 122 patients were included: 36 patients with p16+OPSCC, 16 patients with p16-OPSCC, 4 patients with p16+OCSCC, 41 patients with p16-OCSCC, and 25 healthy controls. The sensitivity and specificity of ddPCR of oropharyngeal swabs against p16 IHC were 92 and 98% respectively, using 20-50 times less RNA than that required for conventional RT-qPCR. Overall agreement between ddPCR of tissue swabs and p16 of tumor tissue was high at ĸ = 0.826 [0.662-0.989]. Oropharyngeal swabs analyzed by ddPCR is a quantitative, rapid, and effective method for minimally invasive oncogenic HPV detection. This assay represents the most sensitive and accurate mode of HPV detection in OPSCC without a tissue biopsy in the available literature.

  9. The Atmospheric Scanning Electron Microscope with open sample space observes dynamic phenomena in liquid or gas

    Energy Technology Data Exchange (ETDEWEB)

    Suga, Mitsuo, E-mail: msuga@jeol.co.jp [Clair Project, JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Nishiyama, Hidetoshi; Konyuba, Yuji [Clair Project, JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Iwamatsu, Shinnosuke; Watanabe, Yoshiyuki [Yamagata Research Institute of Technology, 2-2-1, Matsuei, Yamagata, 990-2473 (Japan); Yoshiura, Chie; Ueda, Takumi [Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan); Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-4, Umezono, Tsukuba 305-8568 (Japan)

    2011-12-15

    Although conventional electron microscopy (EM) requires samples to be in vacuum, most chemical and physical reactions occur in liquid or gas. The Atmospheric Scanning Electron Microscope (ASEM) can observe dynamic phenomena in liquid or gas under atmospheric pressure in real time. An electron-permeable window made of pressure-resistant 100 nm-thick silicon nitride (SiN) film, set into the bottom of the open ASEM sample dish, allows an electron beam to be projected from underneath the sample. A detector positioned below captures backscattered electrons. Using the ASEM, we observed the radiation-induced self-organization process of particles, as well as phenomena accompanying volume change, including evaporation-induced crystallization. Using the electrochemical ASEM dish, we observed tree-like electrochemical depositions on the cathode. In silver nitrate solution, we observed silver depositions near the cathode forming incidental internal voids. The heated ASEM dish allowed observation of patterns of contrast in melting and solidifying solder. Finally, to demonstrate its applicability for monitoring and control of industrial processes, silver paste and solder paste were examined at high throughput. High resolution, imaging speed, flexibility, adaptability, and ease of use facilitate the observation of previously difficult-to-image phenomena, and make the ASEM applicable to various fields. -- Highlights: Black-Right-Pointing-Pointer Atmospheric SEM (ASEM) observes dynamic phenomena in liquid or gas in open ASEM dish. Black-Right-Pointing-Pointer Random motion and radiation-induced self-organization were observed. Black-Right-Pointing-Pointer Tree-like electrochemical deposition of gold was observed on an electrode in situ. Black-Right-Pointing-Pointer Temperature-dependent phase transitions of solder were dynamically observed in air. Black-Right-Pointing-Pointer Silver and solder pastes were easily and rapidly observed in air for process control.

  10. Dengue virus purification and sample preparation for cryo-electron microscopy.

    Science.gov (United States)

    Tan, Joanne L; Lok, Shee Mei

    2014-01-01

    Cryo-electron microscopy (cryo-EM) is a valuable tool used to study the structures of icosahedral viruses without having to resort to crystallization. During the last few decades, significant progress has been made where virus structures previously resolved only to low resolution have now breached the sub-nanometer threshold. Critical to such excellent results are the acquisition of highly purified virus samples and well-frozen samples in vitreous ice. With the virus particles locked in their native conformations, cryo-EM together with single-particle analysis can then be deployed to study the structures of the viruses in their fully hydrated states.

  11. Gas-Assisted Annular Microsprayer for Sample Preparation for Time-Resolved Cryo-Electron Microscopy.

    Science.gov (United States)

    Lu, Zonghuan; Barnard, David; Shaikh, Tanvir R; Meng, Xing; Mannella, Carmen A; Yassin, Aymen; Agrawal, Rajendra; Wagenknecht, Terence; Lu, Toh-Ming

    2014-11-01

    Time-resolved cryo electron microscopy (TRCEM) has emerged as a powerful technique for transient structural characterization of isolated biomacromolecular complexes in their native state within the time scale of seconds to milliseconds. For TRCEM sample preparation, microfluidic device [9] has been demonstrated to be a promising approach to facilitate TRCEM biological sample preparation. It is capable of achieving rapidly aqueous sample mixing, controlled reaction incubation, and sample deposition on electron microscopy (EM) grids for rapid freezing. One of the critical challenges is to transfer samples to cryo-EM grids from the microfluidic device. By using microspraying method, the generated droplet size needs to be controlled to facilitate the thin ice film formation on the grid surface for efficient data collection, while not too thin to be dried out before freezing, i.e., optimized mean droplet size needs to be achieved. In this work, we developed a novel monolithic three dimensional (3D) annular gas-assisted microfluidic sprayer using 3D MEMS (MicroElectroMechanical System) fabrication techniques. The microsprayer demonstrated dense and consistent microsprays with average droplet size between 6-9 μm, which fulfilled the above droplet size requirement for TRCEM sample preparation. With droplet density of around 12-18 per grid window (window size is 58×58 μm), and the data collectible thin ice region of >50% total wetted area, we collected ~800-1000 high quality CCD micrographs in a 6-8 hour period of continuous effort. This level of output is comparable to what were routinely achieved using cryo-grids prepared by conventional blotting and manual data collection. In this case, weeks of data collection process with the previous device [9] has shortened to a day or two. And hundreds of microliter of valuable sample consumption can be reduced to only a small fraction.

  12. Diagnostic performance of swab PCR as an alternative to tissue culture methods for diagnosing infections associated with fracture fixation devices.

    Science.gov (United States)

    Omar, Mohamed; Suero, Eduardo M; Liodakis, Emmanouil; Reichling, Moritz; Guenther, Daniel; Decker, Sebastian; Stiesch, Meike; Krettek, Christian; Eberhard, Jörg

    2016-07-01

    Molecular procedures could potentially improve diagnoses of orthopaedic implant-related infections, but are not yet clinically implemented. Analysis of sonication fluid shows the highest sensitivity for diagnosing implant infections in cases of revision surgery with implant removal. However, there remains controversy regarding the best method for obtaining specimens in cases of revision surgery with implant retention. Tissue culture is the most common diagnostic method for pathogen identification in such cases. Here we aimed to assess the diagnostic performance of swab PCR analysis compared to tissue culture from patients undergoing revision surgery of fracture fixation devices. We prospectively investigated 62 consecutive subjects who underwent revision surgery of fracture fixation devices during a two-year period. Tissue samples were collected for cultures, and swabs from the implant surface were obtained for 16S rRNA PCR analysis. Subjects were classified as having an implant-related infection if (1) they presented with a sinus tract or open wound in communication with the implant; or (2) purulence was encountered intraoperatively; or (3) two out of three tissue cultures tested positive for the presence of the same pathogen. Tissue culture and swab PCR results from the subjects were used to calculate the sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), and area under the ROC curve (AUC) for identifying an orthopaedic implant-related infection. Orthopaedic implant-related infections were detected in 51 subjects. Tissue culture identified infections in 47 cases, and swab PCR in 35 cases. Among the 11 aseptic cases, tissue culture was positive in 2 cases and swab PCR in 4 cases. Tissue culture showed a significantly higher area under the ROC curve for diagnosing infection (AUC=0.89; 95% CI, 0.67-0.96) compared to swab PCR (AUC=0.66; 95% CI, 0.46-0.80) (p=0.033). Compared to swab PCR, tissue culture showed better

  13. Heating-induced variations of secondary electron emission from ion-cleaned copper samples.

    Science.gov (United States)

    Hu, Xiao-Chuan; Zhang, Hai-Bo; Cao, Meng; Zhang, Na; Cui, Wan-Zhao

    2014-09-01

    Secondary electron (SE) emission due to electron impact depends strongly on surface conditions. The variations of SE yield and spectrum with the heating temperature of Ar-ion-cleaned oxygen-free copper samples are therefore measured in situ in a multifunctional ultrahigh vacuum system. The SE yield and the SE spectrum are observed to increase and to narrow, respectively, after sample heating. The maximum SE yield increases from 0.97 before heating to 1.25 after heating at ∼313 °C, and the corresponding full width at half maximum of SE spectrum decreases considerably from 9.3 to 5.5 eV. More CO2 and Ar ions are shown to desorb at a higher heating temperature by residual gas analysis, indicating their contribution to the reduction in work function and surface potential barrier. Ar-ion desorption appears to affect the SE spectrum more than the SE yield. The obtained results provide a new insight into complicated surface influences on SE emission in thermal applications of scanning electron microscopy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron microscopy.

    Science.gov (United States)

    Faas, F G A; Bárcena, M; Agronskaia, A V; Gerritsen, H C; Moscicka, K B; Diebolder, C A; van Driel, L F; Limpens, R W A L; Bos, E; Ravelli, R B G; Koning, R I; Koster, A J

    2013-03-01

    Correlative light and electron microscopy is an increasingly popular technique to study complex biological systems at various levels of resolution. Fluorescence microscopy can be employed to scan large areas to localize regions of interest which are then analyzed by electron microscopy to obtain morphological and structural information from a selected field of view at nm-scale resolution. Previously, an integrated approach to room temperature correlative microscopy was described. Combined use of light and electron microscopy within one instrument greatly simplifies sample handling, avoids cumbersome experimental overheads, simplifies navigation between the two modalities, and improves the success rate of image correlation. Here, an integrated approach for correlative microscopy under cryogenic conditions is presented. Its advantages over the room temperature approach include safeguarding the native hydrated state of the biological specimen, preservation of the fluorescence signal without risk of quenching due to heavy atom stains, and reduced photo bleaching. The potential of cryo integrated light and electron microscopy is demonstrated for the detection of viable bacteria, the study of in vitro polymerized microtubules, the localization of mitochondria in mouse embryonic fibroblasts, and for a search into virus-induced intracellular membrane modifications within mammalian cells. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. Atmospheric scanning electron microscope system with an open sample chamber: Configuration and applications

    Energy Technology Data Exchange (ETDEWEB)

    Nishiyama, Hidetoshi, E-mail: hinishiy@jeol.co.jp [JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Koizumi, Mitsuru, E-mail: koizumi@jeol.co.jp [JEOL Technics Ltd., 2-6-38 Musashino, Akishima, Tokyo 196-0021 (Japan); Ogawa, Koji, E-mail: kogawa@jeol.co.jp [JEOL Technics Ltd., 2-6-38 Musashino, Akishima, Tokyo 196-0021 (Japan); Kitamura, Shinich, E-mail: kitamura@jeol.co.jp [JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Konyuba, Yuji, E-mail: ykonyuub@jeol.co.jp [JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Watanabe, Yoshiyuki, E-mail: watanabeyoshiy@pref.yamagata.jp [Yamagata Research Institute of Technology, 2-2-1, Matsuei, Yamagata 990-2473 (Japan); Ohbayashi, Norihiko, E-mail: n.ohbayashi@m.tohoku.ac.jp [Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, Aobayama, Aoba-ku, Sendai, Miyagi 980-8578 (Japan); Fukuda, Mitsunori, E-mail: nori@m.tohoku.ac.jp [Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, Aobayama, Aoba-ku, Sendai, Miyagi 980-8578 (Japan); Suga, Mitsuo, E-mail: msuga@jeol.co.jp [JEOL Ltd., 3-1-2, Musashino, Akishima, Tokyo 196-8558 (Japan); Sato, Chikara, E-mail: ti-sato@aist.go.jp [Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-4, Umezono, Tsukuba 305-8568 (Japan)

    2014-12-15

    An atmospheric scanning electron microscope (ASEM) with an open sample chamber and optical microscope (OM) is described and recent developments are reported. In this ClairScope system, the base of the open sample dish is sealed to the top of the inverted SEM column, allowing the liquid-immersed sample to be observed by OM from above and by SEM from below. The optical axes of the two microscopes are aligned, ensuring that the same sample areas are imaged to realize quasi-simultaneous correlative microscopy in solution. For example, the cathodoluminescence of ZnO particles was directly demonstrated. The improved system has (i) a fully motorized sample stage, (ii) a column protection system in the case of accidental window breakage, and (iii) an OM/SEM operation system controlled by a graphical user interface. The open sample chamber allows the external administration of reagents during sample observation. We monitored the influence of added NaCl on the random motion of silica particles in liquid. Further, using fluorescence as a transfection marker, the effect of small interfering RNA-mediated knockdown of endogenous Varp on Tyrp1 trafficking in melanocytes was examined. A temperature-regulated titanium ASEM dish allowed the dynamic observation of colloidal silver nanoparticles as they were heated to 240 °C and sintered. - Highlights: • Atmospheric SEM (ASEM) allows observation of samples in liquid or gas. • Open sample chamber allows in situ monitoring of evaporation and sintering processes. • in situ monitoring of processes during reagent administration is also accomplished. • Protection system for film breakage is developed for ASEM. • Usability of ASEM has been improved significantly including GUI control.

  16. Comparison of DRY and WET vaginal swabs with cervical specimens in Roche Cobas 4800 HPV and Abbott RealTime High Risk HPV tests.

    Science.gov (United States)

    Jun, Jae Kwan; Lim, Myong Cheol; Hwang, Sang-Hyun; Shin, Hye Young; Hwang, Na Rae; Kim, Yeon-Jin; Yoo, Chong Woo; Lee, Dong Ock; Joo, Jungnam; Park, Sang-Yoon; Lee, Do-Hoon

    2016-06-01

    Self-collected vaginal swab samples have been proposed as an alternative specimen collection method for human papillomavirus (HPV) DNA detection. Two vaginal swabs (a cone-shaped flocked swab (DRY) and a L-shape FLOQSwab with 2mL eNAT transport medium (WET)) were compared to standard cervical samples for HPV DNA testing. Additionally, they were also compared by using Roche Cobas 4800 HPV (Roche_HPV) and Abbott Real-time High Risk HPV (Abbott_HPV) tests. Ninety-six women were prospectively enrolled from the National Cancer Center in Korea between June and August 2015. WET and DRY vaginal swabs and cervical specimens were collected. Roche_HPV and Abbott_HPV tests were performed. The Roche_HPV test on cervical specimens was used as reference. The observed agreements (kappa) of Roche_HPV and Abbott_HPV between WET and DRY swabs were 89.6% (0.790, 95% confidence interval (95% CI): 0.667-0.913) and 91.7% (0.833, 95%CI: 0.723-0.943), respectively. No statistical difference was observed between WET and DRY swabs (p>0.05 for all comparisons). For HPV16/18, the sensitivity/specificity of Roche_HPV on the DRY and WET samples presented 93.8%/96.3% and 87.5%/97.5%, respectively. For other High Risk HPV (hrHPV), the sensitivity/specificity of Roche_HPV on the DRY and WET swabs presented 91.9%/91.5% and 97.3%/98.3, respectively. The sensitivity/specificity of the Abbott_HPV on the DRY and WET swabs were 93.8%/98.8%, 87.5%/98.8% for HPV16/18, and 91.9%/93.2%, 100.0%/93.2% for other hrHPV, respectively. HPV tests performed similarly when using vaginal DRY and WET swab samples. Using DRY and WET swabs to collect vaginal specimens could be an alternative to collecting cervical samples for HPV DNA testing. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Detection of strep throat causing bacterium directly from medical swabs by touch spray-mass spectrometry.

    Science.gov (United States)

    Jarmusch, Alan K; Pirro, Valentina; Kerian, Kevin S; Cooks, R Graham

    2014-10-07

    Strep throat causing Streptococcus pyogenes was detected in vitro and in simulated clinical samples by performing touch spray ionization-mass spectrometry. MS analysis took only seconds to reveal characteristic bacterial and human lipids. Medical swabs were used as the substrate for ambient ionization. This work constitutes the initial step in developing a non-invasive MS-based test for clinical diagnosis of strep throat. It is limited to the single species, S. pyogenes, which is responsible for the vast majority of cases. The method is complementary to and, with further testing, a potential alternative to current methods of point-of-care detection of S. pyogenes.

  18. Measurement of Sub-Picosecond Electron Bunches via Electro-Optic Sampling of Coherent Transition Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Maxwell, Timothy John [Northern Illinois U.

    2012-01-01

    Future collider applications as well as present high-gradient laser plasma wakefield accelerators and free-electron lasers operating with picosecond bunch durations place a higher demand on the time resolution of bunch distribution diagnostics. This demand has led to significant advancements in the field of electro-optic sampling over the past ten years. These methods allow the probing of diagnostic light such as coherent transition radiation or the bunch wakefields with sub-picosecond time resolution. We present results on the single-shot electro-optic spectral decoding of coherent transition radiation from bunches generated at the Fermilab A0 photoinjector laboratory. A longitudinal double-pulse modulation of the electron beam is also realized by transverse beam masking followed by a transverse-to-longitudinal phase-space exchange beamline. Live profile tuning is demonstrated by upstream beam focusing in conjunction with downstream monitoring of single-shot electro-optic spectral decoding of the coherent transition radiation.

  19. Focussed ion beam thin sample microanalysis using a field emission gun electron probe microanalyser

    Science.gov (United States)

    Kubo, Y.

    2018-01-01

    Field emission gun electron probe microanalysis (FEG-EPMA) in conjunction with wavelength-dispersive X-ray spectrometry using a low acceleration voltage (V acc) allows elemental analysis with sub-micrometre lateral spatial resolution (SR). However, this degree of SR does not necessarily meet the requirements associated with increasingly miniaturised devices. Another challenge related to performing FEG-EPMA with a low V acc is that the accuracy of quantitative analyses is adversely affected, primarily because low energy X-ray lines such as the L- and M-lines must be employed and due to the potential of line interference. One promising means of obtaining high SR with FEG-EPMA is to use thin samples together with high V acc values. This mini-review covers the basic principles of thin-sample FEG-EPMA and describes an application of this technique to the analysis of optical fibres. Outstanding issues related to this technique that must be addressed are also discussed, which include the potential for electron beam damage during analysis of insulating materials and the development of methods to use thin samples for quantitative analysis.

  20. Diverse protocols for correlative super-resolution fluorescence imaging and electron microscopy of chemically fixed samples

    Science.gov (United States)

    Kopek, Benjamin G.; Paez-Segala, Maria G.; Shtengel, Gleb; Sochacki, Kem A.; Sun, Mei G.; Wang, Yalin; Xu, C. Shan; van Engelenburg, Schuyler B.; Taraska, Justin W.; Looger, Loren L.; Hess, Harald F.

    2017-01-01

    Our groups have recently developed related approaches for sample preparation for super-resolution imaging within endogenous cellular environments using correlative light and electron microscopy (CLEM). Four distinct techniques for preparing and acquiring super-resolution CLEM datasets on aldehyde-fixed specimens are provided, including Tokuyasu cryosectioning, whole-cell mount, cell unroofing and platinum replication, and resin embedding and sectioning. Choice of the best protocol for a given application depends on a number of criteria that are discussed in detail. Tokuyasu cryosectioning is relatively rapid but is limited to small, delicate specimens. Whole-cell mount has the simplest sample preparation but is restricted to surface structures. Cell unroofing and platinum replica creates high-contrast, 3-dimensional images of the cytoplasmic surface of the plasma membrane, but is more challenging than whole-cell mount. Resin embedding permits serial sectioning of large samples, but is limited to osmium-resistant probes, and is technically difficult. Expected results from these protocols include super-resolution localization (~10–50 nm) of fluorescent targets within the context of electron microscopy ultrastructure, which can help address cell biological questions. These protocols can be completed in 2–7 days, are compatible with a number of super-resolution imaging protocols, and are broadly applicable across biology. PMID:28384138

  1. Microbial investigations in throat swab and tracheal aspirate specimens are beneficial to predict the corresponding endotracheal tube biofilm flora among intubated neonates with ventilator-associated pneumonia.

    Science.gov (United States)

    Pan, Yun; Du, Lizhong; Ai, Qing; Song, Sijie; Tang, Xiaoli; Zhu, Danping; Yu, Jialin

    2017-08-01

    Ventilator-associated pneumonia (VAP) is a common nosocomial infection in neonatal intensive care units with high morbidity and mortality. Bacterial biofilm in the endotracheal tube (ET) provides a notable and persistent source of pathogens that may cause VAP, and thus is important for VAP detection. However, during intubation microbial investigations in ET, samples are unavailable due to the infeasibility of collecting ET samples during intubation of neonates. It is therefore of great importance to find alternative sources of samples that can help identify the ET biofilm flora. In the present study, the microbial signatures of throat swabs and tracheal aspirates were compared with ET biofilm samples from VAP neonates using 16S ribosomal RNA gene polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE), cloning and sequencing. Sequences were assigned to phylogenetic species using BLAST. Microbial diversity and richness among the three types of specimens were compared based on their DGGE fingerprints, and taxonomic characteristics based on the BLAST results. The microbial richness and diversity of ET biofilms were similar to tracheal aspirate yet significantly different from throat swab samples (P<0.05). Compared with ET biofilms, the overall constituent ratio of microflora was significantly different in throat swab and tracheal aspirate samples (P<0.05). However tracheal aspirate samples were useful for predicting Staphylococcus sp. in ET biofilms with a sensitivity of 85.7% and a specificity of 83.3%. The sensitivity for the combination of tracheal aspirate and throat swab samples to detect Staphylococcus sp. in ET biofilms was 100%. The detection of Pseudomonas sp. in throat swabs assisted its identification in ET biofilms (sensitivity 33.3% and specificity 100%). The results of the present study suggest that microbial investigations in throat swab and tracheal aspirate samples are beneficial for identifying the ET biofilm flora. There may

  2. Detection of Leishmania infantum DNA by real-time PCR in canine oral and conjunctival swabs and comparison with other diagnostic techniques.

    Science.gov (United States)

    Lombardo, Gabriella; Pennisi, Maria Grazia; Lupo, Tiziana; Migliazzo, Antonella; Caprì, Alessandra; Solano-Gallego, Laia

    2012-02-28

    The use of non invasive sampling, such as collection of conjunctival swabs, as a diagnostic tool for the detection of Leishmania DNA is of interest. The purpose of this study was to evaluate the diagnostic utility of detecting Leishmania infection with the use of conjunctival swab samples in dogs living in a highly endemic area for leishmaniosis and to investigate, for the first time, the presence of Leishmania DNA in oral swabs in the same population. One hundred sixty-three dogs living outdoor and recruited in various provinces of Sicily were studied. Leishmania infantum indirect fluorescent antibody test (IFAT), delayed-type hypersensitivity reaction to leishmanin (DTH) and real-time PCR of blood (BL), lymph node (LN), conjunctival (CS) and oral swab (OS) samples were performed. The positive PCR percentages in LN, CS, OS and BL samples were: 24.5%, 22.1%, 8.7% and 5.5%, respectively. Serological and DTH positive percentages were 27.0% and 73.8%, respectively. Seropositive and LN-PCR positive dogs had a high likelihood to be positive by CS-PCR. The similar positive PCR percentages found in CS and LN samples suggest the use of CS-PCR as non-invasive alternative technique to LN-PCR for the detection of Leishmania infection in dogs. In addition, this study demonstrated, for the first time, the presence of Leishmania DNA in oral swabs in dogs. © 2011. Published by Elsevier B.V.

  3. Performance of a Tungsten-Cerium Fluoride Sampling Calorimeter in High-Energy Electron Beam Tests

    CERN Document Server

    Becker, R.; Dissertori, G.; Djambazov, L.; Donega, M.; Lustermann, W.; Marini, A.C.; Nessi-Tedaldi, F.; Pandolfi, F.; Peruzzi, M.; Schönenberger, M.; Cavallari, F.; Dafinei, I.; Diemoz, M.; Lope, C. Jorda; Meridiani, P.; Nuccetelli, M.; Paramatti, R.; Pellegrino, F.; Micheli, F.; Organtini, G.; Rahatlou, S.; Soffi, L.; Brianza, L.; Govoni, P.; Martelli, A.; Tabarelli de Fatis, T.; Monti, V.; Pastrone, N.; Trapani, P.P.; Candelise, V.; Della Ricca, G.

    2015-12-21

    A prototype for a sampling calorimeter made out of cerium fluoride crystals interleaved with tungsten plates, and read out by wavelength-shifting fibres, has been exposed to beams of electrons with energies between 20 and 150 GeV, produced by the CERN Super Proton Synchrotron accelerator complex. The performance of the prototype is presented and compared to that of a Geant4 simulation of the apparatus. Particular emphasis is given to the response uniformity across the channel front face, and to the prototype's energy resolution.

  4. An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology

    Science.gov (United States)

    Thompson, Rebecca F.; Walker, Matt; Siebert, C. Alistair; Muench, Stephen P.; Ranson, Neil A.

    2016-01-01

    Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150 kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a ‘resolution revolution’, owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM. PMID:26931652

  5. Picosecond laser micromachining prior to FIB milling for electronic microscopy sample preparation

    Science.gov (United States)

    Sikora, Aurélien; Fares, Lahouari; Adrian, Jérôme; Goubier, Vincent; Delobbe, Anne; Corbin, Antoine; Sentis, Marc; Sarnet, Thierry

    2017-10-01

    In order to check the manufacturing quality of electronic components using electron microscopy, the area of interest must be exposed. This requires the removal of a large quantity of matter without damaging the surrounding area. This step can be accomplished using ion milling but the processing can last a few hours. In order to accelerate the preparation of the samples, picosecond laser micromachining prior to Focused Ion Beam polishing is envisioned. Laser ablation allows the fast removal of matter but induces damages around the ablated area. Therefore the process has to be optimized in order to limit the size of both the heat affected zone and induced dislocation zone. For this purpose, cavities have been engraved in silicon and in electronic components, using a linearly polarized picosecond laser (∼50 ps) at three different wavelengths (343, 515 and 1030 nm). Results showed that the cross sectional shapes and the surface topologies can be tuned by the laser fluence and the number of pulses. Clear cross sections of bumps and cavity openings, exposing multilayer interfaces, are demonstrated. The silicon removal rates, tuned by the applied energy density, have been measured. Removal rates achieved at 200 kHz were typically hundred times higher than those achieved by ion milling and the best efficiency was obtained at 343 nm.

  6. Characterization of Electron Microscopes with Binary Pseudo-random Multilayer Test Samples

    Energy Technology Data Exchange (ETDEWEB)

    V Yashchuk; R Conley; E Anderson; S Barber; N Bouet; W McKinney; P Takacs; D Voronov

    2011-12-31

    Verification of the reliability of metrology data from high quality X-ray optics requires that adequate methods for test and calibration of the instruments be developed. For such verification for optical surface profilometers in the spatial frequency domain, a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) gratings and arrays has been suggested [1] and [2] and proven to be an effective calibration method for a number of interferometric microscopes, a phase shifting Fizeau interferometer, and a scatterometer [5]. Here we describe the details of development of binary pseudo-random multilayer (BPRML) test samples suitable for characterization of scanning (SEM) and transmission (TEM) electron microscopes. We discuss the results of TEM measurements with the BPRML test samples fabricated from a WiSi2/Si multilayer coating with pseudo-randomly distributed layers. In particular, we demonstrate that significant information about the metrological reliability of the TEM measurements can be extracted even when the fundamental frequency of the BPRML sample is smaller than the Nyquist frequency of the measurements. The measurements demonstrate a number of problems related to the interpretation of the SEM and TEM data. Note that similar BPRML test samples can be used to characterize X-ray microscopes. Corresponding work with X-ray microscopes is in progress.

  7. Characterization of electron microscopes with binary pseudo-random multilayer test samples

    Energy Technology Data Exchange (ETDEWEB)

    Yashchuk, Valeriy V., E-mail: VVYashchuk@lbl.gov [Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Conley, Raymond [NSLS-II, Brookhaven National Laboratory, Upton, NY 11973 (United States); Anderson, Erik H. [Center for X-ray Optics, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Barber, Samuel K. [Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Bouet, Nathalie [NSLS-II, Brookhaven National Laboratory, Upton, NY 11973 (United States); McKinney, Wayne R. [Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States); Takacs, Peter Z. [Brookhaven National Laboratory, Upton, NY 11973 (United States); Voronov, Dmitriy L. [Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 (United States)

    2011-09-01

    Verification of the reliability of metrology data from high quality X-ray optics requires that adequate methods for test and calibration of the instruments be developed. For such verification for optical surface profilometers in the spatial frequency domain, a modulation transfer function (MTF) calibration method based on binary pseudo-random (BPR) gratings and arrays has been suggested and proven to be an effective calibration method for a number of interferometric microscopes, a phase shifting Fizeau interferometer, and a scatterometer [5]. Here we describe the details of development of binary pseudo-random multilayer (BPRML) test samples suitable for characterization of scanning (SEM) and transmission (TEM) electron microscopes. We discuss the results of TEM measurements with the BPRML test samples fabricated from a WiSi{sub 2}/Si multilayer coating with pseudo-randomly distributed layers. In particular, we demonstrate that significant information about the metrological reliability of the TEM measurements can be extracted even when the fundamental frequency of the BPRML sample is smaller than the Nyquist frequency of the measurements. The measurements demonstrate a number of problems related to the interpretation of the SEM and TEM data. Note that similar BPRML test samples can be used to characterize X-ray microscopes. Corresponding work with X-ray microscopes is in progress.

  8. Virtual rough samples to test 3D nanometer-scale scanning electron microscopy stereo photogrammetry.

    Science.gov (United States)

    Villarrubia, J S; Tondare, V N; Vladár, A E

    2016-01-01

    The combination of scanning electron microscopy for high spatial resolution, images from multiple angles to provide 3D information, and commercially available stereo photogrammetry software for 3D reconstruction offers promise for nanometer-scale dimensional metrology in 3D. A method is described to test 3D photogrammetry software by the use of virtual samples-mathematical samples from which simulated images are made for use as inputs to the software under test. The virtual sample is constructed by wrapping a rough skin with any desired power spectral density around a smooth near-trapezoidal line with rounded top corners. Reconstruction is performed with images simulated from different angular viewpoints. The software's reconstructed 3D model is then compared to the known geometry of the virtual sample. Three commercial photogrammetry software packages were tested. Two of them produced results for line height and width that were within close to 1 nm of the correct values. All of the packages exhibited some difficulty in reconstructing details of the surface roughness.

  9. Detection of Leishmania infantum DNA in conjunctival swabs of cats by quantitative real-time PCR.

    Science.gov (United States)

    Benassi, Julia Cristina; Benvenga, Graziella U; Ferreira, Helena Lage; Pereira, Vanessa F; Keid, Lara B; Soares, Rodrigo; Oliveira, Tricia Maria Ferreira de Sousa

    2017-06-01

    Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300-350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. SCANNING ELECTRON MICROSCOPY AND X-RAY DIFFRACTION ANALYSIS OF TANK 18 SAMPLES

    Energy Technology Data Exchange (ETDEWEB)

    Hay, M.; O' Rourke, P.; Ajo, H.

    2012-03-08

    The F-Area Tank Farm (FTF) Performance Assessment (PA) utilizes waste speciation in the waste release model used in the FTF fate and transport modeling. The waste release modeling associated with the residual plutonium in Tank 18 has been identified as a primary contributor to the Tank 18 dose uncertainty. In order to reduce the uncertainty related to plutonium in Tank 18, a better understanding of the plutonium speciation in the Tank 18 waste (including the oxidation state and stoichiometry) is desired. Savannah River National Laboratory (SRNL) utilized Scanning Electron Microscopy (SEM) and X-ray Diffraction (XRD) to analyze Tank 18 samples to provide information on the speciation of plutonium in the waste material. XRD analysis of the Tank 18 samples did not identify any plutonium mineral phases in the samples. These indicates the crystalline mineral phases of plutonium are below the detection limits of the XRD method or that the plutonium phase(s) lack long range order and are present as amorphous or microcrystalline solids. SEM analysis of the Tank 18 samples did locate particles containing plutonium. The plutonium was found as small particles, usually <1 {micro}m but ranging up to several micrometers in diameter, associated with particles of an iron matrix and at low concentration in other elemental matrices. This suggests the plutonium has an affinity for the iron matrix. Qualitatively, the particles of plutonium found in the SEM analysis do not appear to account for all of the plutonium in the sample based on concentrations determined from the chemical analysis of the Tank 18 samples. This suggests that plutonium is also distributed throughout the solids in low concentrations.

  11. Candidate's thesis: the diagnostic utility of an electronic nose: rhinologic applications.

    Science.gov (United States)

    Thaler, Erica R

    2002-09-01

    The thesis explores the applicability of electronic nose technology in medical decision-making. Specifically, the studies undertaken in the thesis were designed to test the ability of the electronic nose to assist in diagnostic questions encountered in the field of rhinology. Three separate studies were undertaken. All involved analysis of specimens by the electronic nose, obtained either in vitro or in vivo: known matched sets of cerebrospinal fluid and serum, bacterial samples from known plated specimens, and culture swabs taken from patients suspected of having rhinosinusitis who also had a matched standard bacterial culture taken from the same site. The goal of analysis was to determine whether the electronic nose was able to identify or categorize specimens or groups of specimens. Each specimen was tested using the organic semiconductor-based Cyranose 320 electronic nose. Data from the 32-element sensor array were subjected to principal-component analysis to depict differences in odorant patterns. Distinction of specimens was identified by calculation of Mahalanobis distance. The electronic nose was able to distinguish serum from cerebrospinal fluid in pure isolates as well as in isolates collected on small cottonoid pledgets at amounts of 0.2 mL or greater. It was also able to distinguish between control swabs and bacterial samples as well as among bacterial samples collected in vitro. Preliminary work suggests that it may be able to distinguish between presence and absence of bacterial infection in specimens collected on nasal swabs. The electronic nose is able to distinguish reliably between cerebrospinal fluid and serum sampled in small amounts, may be able to identify presence and type of bacterial pathogen in vitro, and is able to identify presence or absence of bacteria on nasal swabs. Because this information is available immediately, the electronic nose may be a powerful new technology for diagnostic use, not only for rhinologic purposes but in many

  12. In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate

    Energy Technology Data Exchange (ETDEWEB)

    Barton, B.; Rhinow, D.; Walter, A.; Schroeder, R. [Max Planck Institute of Biophysics, Max-von-Laue Str. 3, 60438 Frankfurt am Main (Germany); Benner, G.; Majorovits, E.; Matijevic, M.; Niebel, H. [Carl Zeiss NTS GmbH, D-73447 Oberkochen (Germany); Mueller, H.; Haider, M. [CEOS GmbH, Englerstr. 26, 69126 Heidleberg (Germany); Lacher, M.; Schmitz, S.; Holik, P. [Caesar Research Center, Ludwig-Erhard-Allee 2, D-53175 Bonn (Germany); Kuehlbrandt, W., E-mail: werner.kuehlbrandt@mpibp-frankfurt.mpg.de [Max Planck Institute of Biophysics, Max-von-Laue Str. 3, 60438 Frankfurt am Main (Germany)

    2011-12-15

    We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, C{sub s} corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of <0.6 nm/h at the specimen. Maximal in-focus phase contrast was obtained by applying voltages between 80 and 700 mV to the phase plate electrode. The phase plate allows for in-focus imaging of biological objects with a signal-to-noise of 5-10 at a resolution of 2-3 nm, as demonstrated for frozen-hydrated virus particles and purple membrane at liquid-nitrogen temperature. -- Highlights: Black-Right-Pointing-Pointer We implement an electrostatic Boersch phase plate into a dedicated prototypical TEM. Black-Right-Pointing-Pointer Phase contrast aberration-corrected electron microscope (PACEM) includes a diffraction magnification unit (DMU). Black-Right-Pointing-Pointer DMU minimizes obstruction of low spatial frequencies by the phase plate. Black-Right-Pointing-Pointer In-focus phase contrast generation is demonstrated for frozen-hydrated biological specimens.

  13. Characteristics of recycled and electron beam irradiated high density polyethylene samples

    Energy Technology Data Exchange (ETDEWEB)

    Cardoso, Jessica R.; Gabriel, Leandro; Geraldo, Aurea B.C.; Moura, Eduardo, E-mail: jrcardoso@ipen.br, E-mail: lgabriell@gmail.com, E-mail: ageraldo@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2015-07-01

    Polymers modification by irradiation is a well-known process that allows degradation and cross-linking in concurrent events; this last is expected when an increase of mechanical properties is required. Actually, the interest of recycling and reuse of polymeric material is linked to the increase of plastics ending up in waste streams. Therefore, these both irradiation and recycling process may be conducted to allow a new use to this material that would be discarded by an improvement of its mechanical properties. In this work, the High Density Polyethylene (HDPE) matrix has been recycled five times from original substrate. The electron beam irradiation process was applied from 50 kGy to 200 kGy in both original and recycled samples; in this way, mechanical properties and thermal characteristics were evaluated. The results of applied process and material characterization are discussed. (author)

  14. In-focus electron microscopy of frozen-hydrated biological samples with a Boersch phase plate.

    Science.gov (United States)

    Barton, B; Rhinow, D; Walter, A; Schröder, R; Benner, G; Majorovits, E; Matijevic, M; Niebel, H; Müller, H; Haider, M; Lacher, M; Schmitz, S; Holik, P; Kühlbrandt, W

    2011-12-01

    We report the implementation of an electrostatic Einzel lens (Boersch) phase plate in a prototype transmission electron microscope dedicated to aberration-corrected cryo-EM. The combination of phase plate, C(s) corrector and Diffraction Magnification Unit (DMU) as a new electron-optical element ensures minimal information loss due to obstruction by the phase plate and enables in-focus phase contrast imaging of large macromolecular assemblies. As no defocussing is necessary and the spherical aberration is corrected, maximal, non-oscillating phase contrast transfer can be achieved up to the information limit of the instrument. A microchip produced by a scalable micro-fabrication process has 10 phase plates, which are positioned in a conjugate, magnified diffraction plane generated by the DMU. Phase plates remained fully functional for weeks or months. The large distance between phase plate and the cryo sample permits the use of an effective anti-contaminator, resulting in ice contamination rates of frozen-hydrated virus particles and purple membrane at liquid-nitrogen temperature. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy

    OpenAIRE

    Can, H?seyin; Caner, Ay?e; D??kaya, Mert; De?irmenci, Aysu; Kara?al?, Sabire; Polat, Ceylan; G?r?z, Y?ksel; ?ner, Ahmet

    2013-01-01

    Background Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone. Material/Methods Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration o...

  16. Detection of Mycoplasma pneumoniae in simulated and true clinical throat swab specimens by nanorod array-surface-enhanced Raman spectroscopy.

    Directory of Open Access Journals (Sweden)

    Suzanne L Hennigan

    Full Text Available The prokaryote Mycoplasma pneumoniae is a major cause of respiratory disease in humans, accounting for 20% of all community-acquired pneumonia and the leading cause of pneumonia in older children and young adults. The limitations of existing options for mycoplasma diagnosis highlight a critical need for a new detection platform with high sensitivity, specificity, and expediency. Here we evaluated silver nanorod arrays (NA as a biosensing platform for detection and differentiation of M. pneumoniae in culture and in spiked and true clinical throat swab samples by surface-enhanced Raman spectroscopy (SERS. Three M. pneumoniae strains were reproducibly differentiated by NA-SERS with 95%-100% specificity and 94-100% sensitivity, and with a lower detection limit exceeding standard PCR. Analysis of throat swab samples spiked with M. pneumoniae yielded detection in a complex, clinically relevant background with >90% accuracy and high sensitivity. In addition, NA-SERS correctly classified with >97% accuracy, ten true clinical throat swab samples previously established by real-time PCR and culture to be positive or negative for M. pneumoniae. Our findings suggest that the unique biochemical specificity of Raman spectroscopy, combined with reproducible spectral enhancement by silver NA, holds great promise as a superior platform for rapid and sensitive detection and identification of M. pneumoniae, with potential for point-of-care application.

  17. A novel quantitative real-time polymerase chain reaction method for detecting toxigenic Pasteurella multocida in nasal swabs from swine.

    Science.gov (United States)

    Scherrer, Simone; Frei, Daniel; Wittenbrink, Max Michael

    2016-12-01

    Progressive atrophic rhinitis (PAR) in pigs is caused by toxigenic Pasteurella multocida. In Switzerland, PAR is monitored by selective culture of nasal swabs and subsequent polymerase chain reaction (PCR) screening of bacterial colonies for the P. multocida toxA gene. A panel of 203 nasal swabs from a recent PAR outbreak were used to evaluate a novel quantitative real-time PCR for toxigenic P. multocida in porcine nasal swabs. In comparison to the conventional PCR with a limit of detection of 100 genome equivalents per PCR reaction, the real-time PCR had a limit of detection of 10 genome equivalents. The real-time PCR detected toxA-positive P. multocida in 101 samples (49.8%), whereas the conventional PCR was less sensitive with 90 toxA-positive samples (44.3%). In comparison to the real-time PCR, 5.4% of the toxA-positive samples revealed unevaluable results by conventional PCR. The approach of culture-coupled toxA PCR for the monitoring of PAR in pigs is substantially improved by a novel quantitative real-time PCR.

  18. Duration of Loxosceles reclusa Venom Detection by ELISA from Swabs

    Science.gov (United States)

    McGLASSON, DAVID L; GREEN, JONATHON A; STOECKER, WILLIAM V; BABCOCK, JAMES L; CALCARA, DAVID A

    2011-01-01

    BACKGROUND Diagnosis of Loxosceles reclusa envenomations is currently based upon clinical presentation. An enzyme-linked immunosorbent assay (ELISA) can detect surface Loxosceles venom at the envenomation site, allowing diagnostic confirmation. The length of time that venom on the skin is recoverable non-invasively is unknown. MATERIALS AND METHODS To investigate duration of recoverable venom antigen, whole venom and fractionated sphingomyelinase D venom aliquots were injected subcutaneously in New Zealand White rabbits. Cotton and Dacron swabs were compared for venom recovery over a 21-day period using a surface swab technique. RESULTS Significant amounts of Loxosceles reclusa antigen were found on the surface of rabbit skin after experimental injection of whole venom and sphingomyelinase D. The duration of recoverable antigen using this experimental model appears to be at least two weeks and as long as 21 days in some cases. CONCLUSIONS Because the duration of the recoverable antigen is seen to be at least two weeks, the ELISA venom test appears capable of detecting venom on most patients presenting with Loxosceles envenomations. This detection system will allow the physician more accurate determination of whether the lesion is from a brown recluse spider or some other agent that can cause this type of necrotic ulcer. PMID:19967916

  19. Duration of Loxosceles reclusa venom detection by ELISA from swabs.

    Science.gov (United States)

    McGlasson, David L; Green, Jonathon A; Stoecker, William V; Babcock, James L; Calcara, David A

    2009-01-01

    Diagnosis of Loxosceles reclusa envenomations is currently based upon clinical presentation. An enzyme-linked immunosorbent assay (ELISA) can detect surface Loxosceles venom at the envenomation site, allowing diagnostic confirmation. The length of time that venom on the skin is recoverable non-invasively is unknown. To investigate duration of recoverable venom antigen, whole venom and fractionated sphingomyelinase D venom aliquots were injected subcutaneously in New Zealand White rabbits. Cotton and Dacron swabs were compared for venom recovery over a 21-day period using a surface swab technique. Significant amounts of Loxosceles reclusa antigen were found on the surface of rabbit skin after experimental injection of whole venom and sphingomyelinase D. The duration of recoverable antigen using this experimental model appears to be at least two weeks and as long as 21 days in some cases. Because the duration of the recoverable antigen is seen to be at least two weeks, the ELISA venom test appears capable of detecting venom on most patients presenting with Loxosceles envenomations. This detection system will allow the physician more accurate determination of whether the lesion is from a brown recluse spider or some other agent that can cause this type of necrotic ulcer.

  20. A study of female genital swabs in primary health care centres in Jos, Nigeria

    Directory of Open Access Journals (Sweden)

    Samuel Nwadioha

    2011-03-01

    Full Text Available Objective: To detect some common microbial agents of female genital discharges in order to improve the current syndromic management of abnormal vaginal discharge. Methods: A prospective study of female genital swabs collected from Primary Health Care Centres, Jos, and analysed for microscopy, culture and sensitivity in Jos University Teaching Hospital, December 2006 to December 2007 was carried out. Results: Microbial agents were detected in 70% (700 of a total 1 000 female genital swabs studied. Candida species peaked with 42.0% (420 out of the 1000 samples, followed by Gardnerella vaginalis, an agent of bacterial vaginosis with 26.0%. The distribution of abnormal vaginal discharge was highest in young adults aged 21 to 30 years. Conclusions: It is concluded that abnormal vaginal discharge is most prevalent in the young sexually active age group with Candida species as the commonest agent. We recommend prevention, early diagnosis and prompt treatment of infective female genital discharge in order to reduce the menace of HIV transmission.

  1. Lights Will Guide You : Sample Preparation and Applications for Integrated Laser and Electron Microscopy

    Science.gov (United States)

    Karreman, M. A.

    2013-03-01

    Correlative microscopy is the combined use of two different forms of microscopy in the study of a specimen, allowing for the exploitation of the advantages of both imaging tools. The integrated Laser and Electron Microscope (iLEM), developed at Utrecht University, combines a fluorescence microscope (FM) and a transmission electron microscope (TEM) in a single set-up. The region of interest in the specimen is labeled or tagged with a fluorescent probe and can easily be identified within a large field of view with the FM. Next, this same area is retraced in the TEM and can be studied at high resolution. The iLEM demands samples that can be imaged with both FM and TEM. Biological specimen, typically composed of light elements, generate low image contrast in the TEM. Therefore, these samples are often ‘contrasted’ with heavy metal stains. FM, on the other hand, images fluorescent samples. Sample preparation for correlative microscopy, and iLEM in particular, is complicated by the fact that the heavy metals stains employed for TEM quench the fluorescent signal of the probe that is imaged with FM. The first part of this thesis outlines preparation procedures for biological material yielding specimen that can be imaged with the iLEM. Here, approaches for the contrasting of thin sections of cells and tissue are introduced that do not affect the fluorescence signal of the probe that marks the region of interest. Furthermore, two novel procedures, VIS2FIXH and VIS2FIX­FS are described that allow for the chemical fixation of thin sections of cryo-immobilized material. These procedures greatly expedite the sample preparation process, and open up novel possibilities for the immuno-labeling of difficult antigens, eg. proteins and lipids that are challenging to preserve. The second part of this thesis describes applications of iLEM in research in the field of life and material science. The iLEM was employed in the study of UVC induced apoptosis (programmed cell death) of

  2. Prevalence of mycoplasmas in the semen and vaginal swabs of Danish stallions and mares

    DEFF Research Database (Denmark)

    Baczynska, Agata; Fedder, Jens; Schougaard, Hans

    2007-01-01

    and semen of stallions, showed the presence of different Mycoplasma species. Therefore our study aimed to find the prevalence of Mycoplasma species and a possible association with fertility problems in Danish riding horses. Eighty semen samples from stallions and 19 vaginal swab samples from mares were......The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections...... may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis...

  3. Prevalence of mycoplasmas in the semen and vaginal swabs of Danish stallions and mares

    DEFF Research Database (Denmark)

    Baczynska, Agata; Fedder, J.; Schougaard, H.

    2006-01-01

    and semen of stallions, showed the presence of different Mycoplasma species. Therefore our study aimed to find the prevalence of Mycoplasma species and a possible association with fertility problems in Danish riding horses. Eighty semen samples from stallions and 19 vaginal swab samples from mares were......The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections...... may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis...

  4. Comparison of three methods for the recovery of skin pathogens from impetigo swabs collected in a remote community of Northern Territory, Australia.

    Science.gov (United States)

    Bowen, Asha C; Tong, Steven Y C; Chatfield, Mark D; Andrews, Ross M; Carapetis, Jonathan R

    2013-06-01

    Impetigo is a common infection in children living in remote areas. Immediate plating of impetigo swabs is the gold standard for bacterial recovery but is rarely feasible in remote regions. Bacterial culture increases our understanding of antibiotic resistance and strain diversity, which guides treatment protocols and epidemiological monitoring. We investigated three practical alternatives for recovering Streptococcus pyogenes and Staphylococcus aureus from transported swabs: dry swabs transported at 4°C with desiccant and plated within 48 h; swabs inoculated into skim milk tryptone glucose glycerol broth (STGGB), transported at 4°C, stored at -70°C and plated within 61 days; and ESwabs inoculated into Amies broth, transported at 4°C and plated within 48 h. Detection of Strep. pyogenes and Staph. aureus from simultaneously collected swabs was compared for the dry vs STGGB (36 sores) and the STGGB vs Amies (39 sores) methods. Swabs were collected from 43 children (75 sores sampled) in a remote community of Northern Territory, Australia in November 2011. The children had impetigo and were participating in the Skin Sore Trial [Australian Clinical Trials Registry ACTRN12609000858291]. Recovery of Strep. pyogenes for dry vs STGGB was 72% (26/36) and 92% (33/36) and for STGGB vs Amies was 92% (36/39) for both methods. Staphylococcus aureus recovery for dry vs STGGB was 69% (25/36) and 72% 26/36) and for STGGB vs Amies was 74% (29/39) and 85% (33/39). STGGB and Amies media provided higher recovery of Strep. pyogenes than dry swabs. These results and the opportunity to batch and store specimens for molecular studies support the use of STGGB transport media for future impetigo research.

  5. Sampling

    CERN Document Server

    Thompson, Steven K

    2012-01-01

    Praise for the Second Edition "This book has never had a competitor. It is the only book that takes a broad approach to sampling . . . any good personal statistics library should include a copy of this book." —Technometrics "Well-written . . . an excellent book on an important subject. Highly recommended." —Choice "An ideal reference for scientific researchers and other professionals who use sampling." —Zentralblatt Math Features new developments in the field combined with all aspects of obtaining, interpreting, and using sample data Sampling provides an up-to-date treat

  6. Contribution of the Ogawa-Kudoh swab culture method to the diagnosis of pulmonary tuberculosis in Brazil.

    Science.gov (United States)

    Palaci, M; Peres, R L; Maia, R; Cunha, E A T; Ribeiro, M O; Lecco, R; de Souza Ribeiro, C; Ferro E Silva, R R; Vinhas, S A; Dietze, R; Vianna, S; de Morais, C G V

    2013-06-01

    To analyse the contribution of the Ogawa-Kudoh (O-K) swab culture method to the diagnosis of pulmonary tuberculosis (PTB) in four different regions of Brazil. This study was carried out in two phases. Phase 1 was designed to compare the direct swab culture method (O-K) with the culture concentrated method (N-acetyl-L-cysteine-sodium hydroxide [NALC-NaOH]); for this purpose, 569 sputum samples were cultured by both methods. Phase 2 was carried out to assess the contribution of the O-K method to the diagnosis of PTB in four different regions in Brazil, based on the evaluation of 19,163 sputum samples. In the first phase of the study, O-K culture had a sensitivity of 94.8% and specificity of 99.8% in cases confirmed by NALC-NaOH/Löwenstein-Jensen (LJ) culture. In the second phase of the study, the overall contribution of O-K culture compared to acid-fast bacilli (AFB) examination (AFB-/culture+) to the diagnosis of PTB was 29.8%. O-K culture contributes significantly to the diagnosis of smear-negative PTB. Importantly, this method allows the recovery of clinical isolates in areas where use of the standard culture centrifuge is impossible, indicating that the O-K swab culture method should become a standard method for TB diagnosis in these regions.

  7. Electronic Cigarettes: Awareness, Recent Use, and Attitudes Within a Sample of Socioeconomically Disadvantaged Australian Smokers.

    Science.gov (United States)

    Twyman, Laura; Bonevski, Billie; Paul, Christine; Bryant, Jamie; Gartner, Coral; Guillaumier, Ashleigh

    2016-05-01

    Electronic cigarette (e-cigarette) awareness, trial of e-cigarettes in the past 12 months, source and perceptions of safety and effectiveness was assessed within a disadvantaged sample of adult Australian smokers receiving welfare aid. A cross-sectional survey was administered to clients who smoke at two community service organizations in New South Wales, Australia from October 2013 to July 2014. E-cigarette awareness, trial in past 12 months, sources of e-cigarettes and perceptions of the safety and effectiveness of e-cigarettes to help people quit were assessed along with sociodemographic and smoking-related variables. In total, 369 participants completed the survey (77% response rate). Awareness and trial of e-cigarettes were reported by 77% (n = 283) and 35% (n = 103) of the sample, respectively. E-cigarettes were most commonly obtained from friends/strangers followed by tobacco shops (tobacconists). Trying e-cigarettes in the past 12 months was significantly associated with positive perceptions of their safety (odds ratio [OR] = 1.8, 95% confidence interval [CI] = 1, 3.1) and effectiveness (OR = 1.9, 95% CI = 1.1, 3.2). Motivation to quit tobacco smoking was also significantly positively associated with positive perceptions of e-cigarette safety (OR = 1.2, 95% CI = 1.1, 1.4) and effectiveness (OR = 1.2, 95% CI = 1.0, 1.3). Rates of awareness and trial of e-cigarettes within a disadvantaged sample of Australian smokers are comparable to rates found within representative samples of the general Australian population. Previously trying e-cigarettes and higher levels of motivation to quit were associated with more positive perceptions of e-cigarette safety and effectiveness. This study demonstrates that socioeconomically disadvantaged smokers are aware of and accessing e-cigarettes in a country with relatively high restrictions covering e-cigarette sale and use. © The Author 2015. Published by Oxford University Press on behalf of the Society for Research on

  8. Space-time sampling strategies for electronically steerable incoherent scatter radar

    Science.gov (United States)

    Swoboda, John

    ncoherent scatter radar (ISR) systems allow researchers to peer into the ionosphere via remote sensing of intrinsic plasma parameters. ISR sensors have been used since the 1950s and until the past decade were mainly equipped with a single mechanically steerable antenna. As such, the ability to develop a two or three dimensional picture of the plasma parameters in the ionosphere has been constrained by the relatively slow mechanical steering of the antennas. A newer class of systems using electronically steerable array (ESA) antennas have broken the chains of this constraint, allowing researchers to create 3-D reconstructions of plasma parameters. There have been many studies associated with reconstructing 3-D fields of plasma parameters, but there has not been a systematic analysis into the sampling issues that arise. Also, there has not been a systematic study as to how to reconstruct these plasma parameters in an optimum sense as opposed to just using different forms of interpolation. The research presented here forms a framework that scientists and engineers can use to plan experiments with ESA ISR capabilities and to better analyze the resulting data. This framework attacks the problem of space-time sampling by ESA ISR systems from the point of view of signal processing, simulation and inverse theoretic image reconstruction. We first describe a physics based model of incoherent scatter from the ionospheric plasma, along with processing methods needed to create the plasma parameter measurements. Our approach leads to development of the space-time ambiguity function, forming a theoretical foundation of the forward model for ISR. This forward model is novel in that it takes into account the shape of the antenna beam and scanning method along with integration time to develop the proper statistics for a desired measurement precision. Once the forward model is developed, we present the simulation method behind the Simulator for ISR (SimISR). SimISR uses input plasma

  9. Electronic noses for monitoring benzene occupational exposure in biological samples of Egyptian workers

    Directory of Open Access Journals (Sweden)

    Ehab I. Mohamed

    2013-02-01

    Full Text Available Objectives: Benzene is commonly emitted in several industries, leading to widespread environmental and occupational exposure hazards. While less toxic solvents have been substituted for benzene, it is still a component of petroleum products and is a trace impurity in industrial products resulting in continued higher occupational exposures in industrial settings in developing countries. Materials and Methods: We investigated the potential use of an electronic nose (e-nose to monitor the headspace volatiles in biological samples from benzene-exposed Egyptian workers and non-exposed controls. The study population comprised 150 non-smoking male workers exposed to benzene and an equal number of matching non-exposed controls. We determined biomarkers of benzene used to estimate exposure and risk including: benzene in exhaled air and blood; and its urinary metabolites such as phenol and muconic acid using gas chromatography technique and a portable e-nose. Results: The average benzene concentration measured in the ambient air of the workplace of all studied industrial settings in Alexandria, Egypt; was 97.56±88.12 μg/m3 (range: 4.69–260.86 μg/m3. Levels of phenol and muconic acid were signifi cantly (p < 0.001 higher in both blood and urine of benzene-exposed workers as compared to non-exposed controls. Conclusions: The e-nose technology has successfully classifi ed and distinguished benzene-exposed workers from non-exposed controls for all measured samples of blood, urine and the exhaled air with a very high degree of precision. Thus, it will be a very useful tool for the low-cost mass screening and early detection of health hazards associated with the exposure to benzene in the industry.

  10. Actinomyces hominis sp. nov., isolated from a wound swab.

    Science.gov (United States)

    Funke, Guido; Englert, Ralf; Frodl, Reinhard; Bernard, Kathryn A; Stenger, Steffen

    2010-07-01

    A coryneform bacterium (strain 1094(T)) was isolated from a wound swab taken from an 89-year-old female patient. Chemotaxonomic investigations suggested that this bacterium was related to the genera Actinomyces, Arcanobacterium and Actinobaculum. Phylogenetic analysis of 16S rRNA gene sequences showed that strain 1094(T) was most closely related to Actinomyces europaeus CCUG 32789 A(T) (94.3 % similarity). Phenotypically, the isolate could be separated from its closest phylogenetic neighbours on the basis of being positive for catalase, CAMP reaction, acid phosphatase, N-acetyl-beta-glucosaminidase and raffinose fermentation. Based on the data presented, it is proposed that strain 1094(T) should be classified in a novel species, Actinomyces hominis sp. nov. The type strain is 1094(T) (=CCUG 57540(T) =DSM 22168(T)).

  11. Effect of lingual gauze swab placement on pulse oximeter readings in anaesthetised dogs and cats.

    Science.gov (United States)

    Mair, A; Martinez-Taboada, F; Nitzan, M

    2017-01-14

    This study aimed to evaluate the effect of lingual gauze swab placement on pulse oximeter readings in anaesthetised dogs and cats. Following anaesthetic induction, the following pulse oximeter probe configurations were performed: no gauze swab (control), placement of a gauze swab between the tongue and the probe, placement of different thicknesses of gauze swab, placement of red cotton fabric, placement of a sheet of white paper and placement of the probe and gauze swab on different locations on the tongue. Oxygen saturation (SpO2) and peripheral perfusion index (PI) were recorded. Placement of a gauze swab between the pulse oximeter probe and the tongue in anaesthetised dogs and cats resulted in significantly higher SpO2 values compared with the control group. In dogs, PI values were significantly higher than the control in all groups except the quarter thickness swab group. In cats, PI was significantly higher in the double thickness swab and white paper groups compared with the control. Cats had significantly higher SpO2 and lower PI values than dogs. The authors propose that increased contact pressure is responsible for significantly higher SpO2 and PI readings with the use of a lingual gauze swab resulting from changes in transmural pressure and arterial compliance. British Veterinary Association.

  12. Equivalence of self- and staff-collected nasal swabs for the detection of viral respiratory pathogens.

    Directory of Open Access Journals (Sweden)

    Manas K Akmatov

    Full Text Available BACKGROUND: The need for the timely collection of diagnostic biosamples during symptomatic episodes represents a major obstacle to large-scale studies on acute respiratory infection (ARI epidemiology. This may be circumvented by having the participants collect their own nasal swabs. We compared self- and staff-collected swabs in terms of swabbing quality and detection of viral respiratory pathogens. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a prospective study among employees of our institution during the ARI season 2010/2011 (December-March. Weekly emails were sent to the participants (n = 84, reminding them to come to the study center in case of new symptoms. The participants self-collected an anterior nasal swab from one nostril, and trained study personnel collected one from the other nostril. The participants self-collected another two swabs (one from each nostril on a subsequent day. Human β-actin DNA concentration was determined in the swabs as a quality control. Viral respiratory pathogens were detected by multiplex RT-PCR (Seeplex RV15 kit, Seegene, Eschborn, Germany. Of 84 participants, 56 (67% reported at least one ARI episode, 18 participants two, and one participant three. Self-swabbing was highly accepted by the participants. The amount of β-actin DNA per swab was higher in the self- than in the staff-collected swabs (p = 0.008. β-actin concentration was lower in the self-swabs collected on day 1 than in those collected on a subsequent day (p<0.0001. A respiratory viral pathogen was detected in 31% (23/75 of staff- and in 35% (26/75 of self-collected swabs (p = 0.36. With both approaches, the most frequently identified pathogens were human rhinoviruses A/B/C (12/75 swabs, 16% and human coronavirus OC43 (4/75 swabs, 5%. There was almost perfect agreement between self- and staff-collected swabs in terms of pathogen detection (agreement = 93%, kappa = 0.85, p<0.0001. CONCLUSIONS/SIGNIFICANCE: Nasal self-swabbing

  13. Equivalence of self- and staff-collected nasal swabs for the detection of viral respiratory pathogens.

    Science.gov (United States)

    Akmatov, Manas K; Gatzemeier, Anja; Schughart, Klaus; Pessler, Frank

    2012-01-01

    The need for the timely collection of diagnostic biosamples during symptomatic episodes represents a major obstacle to large-scale studies on acute respiratory infection (ARI) epidemiology. This may be circumvented by having the participants collect their own nasal swabs. We compared self- and staff-collected swabs in terms of swabbing quality and detection of viral respiratory pathogens. We conducted a prospective study among employees of our institution during the ARI season 2010/2011 (December-March). Weekly emails were sent to the participants (n = 84), reminding them to come to the study center in case of new symptoms. The participants self-collected an anterior nasal swab from one nostril, and trained study personnel collected one from the other nostril. The participants self-collected another two swabs (one from each nostril) on a subsequent day. Human β-actin DNA concentration was determined in the swabs as a quality control. Viral respiratory pathogens were detected by multiplex RT-PCR (Seeplex RV15 kit, Seegene, Eschborn, Germany). Of 84 participants, 56 (67%) reported at least one ARI episode, 18 participants two, and one participant three. Self-swabbing was highly accepted by the participants. The amount of β-actin DNA per swab was higher in the self- than in the staff-collected swabs (p = 0.008). β-actin concentration was lower in the self-swabs collected on day 1 than in those collected on a subsequent day (p<0.0001). A respiratory viral pathogen was detected in 31% (23/75) of staff- and in 35% (26/75) of self-collected swabs (p = 0.36). With both approaches, the most frequently identified pathogens were human rhinoviruses A/B/C (12/75 swabs, 16%) and human coronavirus OC43 (4/75 swabs, 5%). There was almost perfect agreement between self- and staff-collected swabs in terms of pathogen detection (agreement = 93%, kappa = 0.85, p<0.0001). Nasal self-swabbing for identification of viral ARI pathogens proved to be equivalent to

  14. A multi-sample changer coupled to an electron cyclotron resonance source for accelerator mass spectrometry experiments.

    Science.gov (United States)

    Vondrasek, R; Palchan, T; Pardo, R; Peters, C; Power, M; Scott, R

    2014-02-01

    A new multi-sample changer has been constructed allowing rapid changes between samples. The sample changer has 20 positions and is capable of moving between samples in 1 min. The sample changer is part of a project using Accelerator Mass Spectrometry (AMS) at the Argonne Tandem Linac Accelerator System (ATLAS) facility to measure neutron capture rates on a wide range of actinides in a reactor environment. This project will require the measurement of a large number of samples previously irradiated in the Advanced Test Reactor at Idaho National Laboratory. The AMS technique at ATLAS is based on production of highly charged positive ions in an electron cyclotron resonance ion source followed by acceleration in the ATLAS linac. The sample material is introduced into the plasma via laser ablation chosen to limit the dependency of material feed rates upon the source material composition as well as minimize cross-talk between samples.

  15. Rapid diagnosis of diarrhea caused by Shigella sonnei using dipsticks; comparison of rectal swabs, direct stool and stool culture.

    Directory of Open Access Journals (Sweden)

    Claudia Duran

    Full Text Available BACKGROUND: We evaluated a dipstick test for rapid detection of Shigella sonnei on bacterial colonies, directly on stools and from rectal swabs because in actual field situations, most pathologic specimens for diagnosis correspond to stool samples or rectal swabs. METHODOLOGY/PRINCIPAL FINDINGS: The test is based on the detection of S. sonnei lipopolysaccharide (LPS O-side chains using phase I-specific monoclonal antibodies coupled to gold particles, and displayed on a one-step immunochromatographic dipstick. A concentration as low as 5 ng/ml of LPS was detected in distilled water and in reconstituted stools in 6 minutes. This is the optimal time for lecture to avoid errors of interpretation. In distilled water and in reconstituted stools, an unequivocal positive reaction was obtained with 4 x 10(6 CFU/ml of S. sonnei. The specificity was 100% when tested with a battery of Shigella and different unrelated strains. When tested on 342 rectal swabs in Chile, specificity (281/295 was 95.3% (95% CI: 92.9% - 97.7% and sensitivity (47/47 was 100%. Stool cultures and the immunochromatographic test showed concordant results in 95.5 % of cases (328/342 in comparative studies. Positive and negative predictive values were 77% (95% CI: 65% - 86.5% and 100% respectively. When tested on 219 stools in Chile, Vietnam, India and France, specificity (190/198 was 96% (95% CI 92%-98% and sensitivity (21/21 was 100%. Stool cultures and the immunochromatographic test showed concordant results in 96.3 % of cases (211/219 in comparative studies. Positive and negative predictive values were 72.4% (95% CI 56.1%-88.6% and 100 %, respectively. CONCLUSION: This one-step dipstick test performed well for diagnosis of S. sonnei both on stools and on rectal swabs. These data confirm a preliminary study done in Chile.

  16. Closer to the native state. Critical evaluation of cryo-techniques for Transmission Electron Microscopy: preparation of biological samples.

    Science.gov (United States)

    Mielanczyk, Lukasz; Matysiak, Natalia; Michalski, Marek; Buldak, Rafal; Wojnicz, Romuald

    2014-01-01

    Over the years Transmission Electron Microscopy (TEM) has evolved into a powerful technique for the structural analysis of cells and tissues at various levels of resolution. However, optimal sample preservation is required to achieve results consistent with reality. During the last few decades, conventional preparation methods have provided most of the knowledge about the ultrastructure of organelles, cells and tissues. Nevertheless, some artefacts can be introduced at all stagesofstandard electron microscopy preparation technique. Instead, rapid freezing techniques preserve biological specimens as close as possible to the native state. Our review focuses on different cryo-preparation approaches, starting from vitrification methods dependent on sample size. Afterwards, we discuss Cryo-Electron Microscopy Of VItreous Sections (CEMOVIS) and the main difficulties associated with this technique. Cryo-Focused Ion Beam (cryo-FIB) is described as a potential alternative for CEMOVIS. Another post-processing route for vitrified samples is freeze substitution and embedding in resin for structural analysis or immunolocalization analysis. Cryo-sectioning according to Tokuyasu is a technique dedicated to high efficiency immunogold labelling. Finally, we introduce hybrid techniques, which combine advantages of primary techniques originally dedicated to different approaches. Hybrid approaches permit to perform the study of difficult-to-fix samples and antigens or help optimize the sample preparation protocol for the integrated Laser and Electron Microscopy (iLEM) technique.

  17. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-03-30

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  18. U-10Mo Sample Preparation and Examination using Optical and Scanning Electron Microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Prabhakaran, Ramprashad [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Joshi, Vineet V. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rhodes, Mark A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schemer-Kohrn, Alan L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Guzman, Anthony D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lavender, Curt A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2016-10-01

    The purpose of this document is to provide guidelines to prepare specimens of uranium alloyed with 10 weight percent molybdenum (U-10Mo) for optical metallography and scanning electron microscopy. This document also provides instructions to set up an optical microscope and a scanning electron microscope to analyze U-10Mo specimens and to obtain the required information.

  19. Localization of fluorescently labeled structures in frozen-hydrated samples using integrated light electron microscopy

    NARCIS (Netherlands)

    Faas, F.G.A.; Bárcena, M.A.; Agronskaia, A.V.; Gerritsen, H.C.; Moscicka, K.B.; Diebolder, C.A.; Driel, L.F.; Limpens, R.W.A.L.; Bos, E.; Ravelli, R.B.G.; Koning, R.I.; Koster, A.J.

    2013-01-01

    Correlative light and electron microscopy is an increasingly popular technique to study complex biological systems at various levels of resolution. Fluorescence microscopy can be employed to scan large areas to localize regions of interest which are then analyzed by electron microscopy to obtain

  20. Note on in situ (scanning) transmission electron microscopy study of liquid samples.

    Science.gov (United States)

    Jiang, Nan

    2017-08-01

    Liquid cell (scanning) transmission electron microscopy has been developed rapidly, using amorphous SiNx membranes as electron transparent windows. The current interpretations of electron beam effects are mainly based on radiolytic processes. In this note, additional effects of the electric field due to electron-beam irradiation are discussed. The electric field can be produced by the charge accumulation due to the emission of secondary and Auger electrons. Besides various beam-induced phenomena, such as nanoparticle precipitation and gas bubble formation and motion, two other effects need to be considered; one is the change of Gibbs free energy of nucleation and the other is the violation of Brownian motion due to ion drifting driven by the electric field. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Revised SWAB guidelines for antimicrobial therapy of community-acquired pneumonia

    NARCIS (Netherlands)

    Schouten, JA; Prins, JM; Bonten, MJ; Degener, J; Janknegt, RE; Hollander, JMR; Jonkers, RE; Wijnands, WJ; Verheij, TJ; Sachs, APE; Kullberg, BJ

    The Dutch Working Party on Antibiotic Policy (SWAB) develops evidence-based guidelines, aimed at optimalisation of antibiotic use and limitation of the spread of antimicrobial resistance. A revision of the SWAB guideline for the treatment of community-acquired pneumonia (CAP), published in 1998, was

  2. The application of alkaline lysis and pressure cycling technology in the differential extraction of DNA from sperm and epithelial cells recovered from cotton swabs.

    Science.gov (United States)

    Nori, Deepthi V; McCord, Bruce R

    2015-09-01

    This study reports the development of a two-step protocol using pressure cycling technology (PCT) and alkaline lysis for differential extraction of DNA from mixtures of sperm and vaginal epithelial cells recovered from cotton swabs. In controlled experiments, in which equal quantities of sperm and female epithelial cells were added to cotton swabs, 5 min of pressure pulsing in the presence of 0.4 M NaOH resulted in 104 ± 6% recovery of female epithelial DNA present on the swab. Following the pressure treatment, exposing the swabs to a second 5-min alkaline treatment at 95 °C without pressure resulted in the selective recovery of 69 ± 6% of the sperm DNA. The recovery of the vaginal epithelia and sperm DNA was optimized by examining the effect of sodium hydroxide concentration, incubation temperature, and time. Following the alkaline lysis steps, the samples were neutralized with 2 M Tris (pH 7.5) and purified with phenol-chloroform-isoamyl alcohol to permit downstream analysis. The total processing time to remove both fractions from the swab was less than 20 min. Short tandem repeat (STR) analysis of these fractions obtained from PCT treatment and alkaline lysis generated clean profiles of female epithelial DNA and male sperm DNA for 1:1 mixtures of female and male cells and predominant male profiles for mixtures up to 5:1 female to male cells. By reducing the time and increasing the recovery of DNA from cotton swabs, this new method presents a novel and potentially useful procedure for forensic differential extractions.

  3. A Miniaturized Variable Pressure Scanning Electron Microscope (MVP-SEM) for In-Situ Mars Surface Sample Analysis

    Science.gov (United States)

    Edmunson, J.; Gaskin, J. A.; Jerman, G. A.; Harvey, R. P.; Doloboff, I. J.; Neidholdt, E. L.

    2016-01-01

    The Miniaturized Variable Pressure Scanning Electron Microscope (MVP-SEM) project, funded by the NASA Planetary Instrument Concepts for the Advancement of Solar System Observations (PICASSO) Research Opportunities in Space and Earth Sciences (ROSES), will build upon previous miniaturized SEM designs and recent advancements in variable pressure SEM's to design and build a SEM to complete analyses of samples on the surface of Mars using the atmosphere as an imaging medium. This project is a collaboration between NASA Marshall Space Flight Center (MSFC), the Jet Propulsion Laboratory (JPL), electron gun and optics manufacturer Applied Physics Technologies, and small vacuum system manufacturer Creare. Dr. Ralph Harvery and environmental SEM (ESEM) inventor Dr. Gerry Danilatos serve as advisors to the team. Variable pressure SEMs allow for fine (nm-scale) resolution imaging and micron-scale chemical study of materials without sample preparation (e.g., carbon or gold coating). Charging of a sample is reduced or eliminated by the gas surrounding the sample. It is this property of ESEMs that make them ideal for locations where sample preparation is not yet feasible, such as the surface of Mars. In addition, the lack of sample preparation needed here will simplify the sample acquisition process and allow caching of the samples for future complementary payload use.

  4. Efficiency and limitation of periodic sample multiplication to reduce computational load in Monte Carlo simulations of electron swarms in gas under attachment-dominated conditions

    Science.gov (United States)

    Sugawara, Hirotake

    2018-03-01

    In Monte Carlo simulations of electron swarms, sample electrons were copied periodically so that a sufficient number of samples are obtained in equilibrium after relaxation even under a severe attachment-dominated condition where most electrons vanish during the relaxation. The final sampling results were equivalent to those sampled by a conventional method, and the computational time conventionally wasted for the tracking of vanishing electrons was reduced drastically. The time saved can be utilized for tracking more samples to reduce statistical fluctuation. The efficiency of this technique and its limitation are discussed quantitatively together with details on its implementation.

  5. Measurement of electron spin-lattice relaxation times in radical doped butanol samples at 1 K using the NEDOR method

    Energy Technology Data Exchange (ETDEWEB)

    Hess, C., E-mail: hess@ep1.rub.de [Ruhr-Universitaet Bochum, Experimentalphysik I, Universitaetsstr. 150, 44801 Bochum (Germany); Herick, J.; Berlin, A.; Meyer, W.; Reicherz, G. [Ruhr-Universitaet Bochum, Experimentalphysik I, Universitaetsstr. 150, 44801 Bochum (Germany)

    2012-12-01

    The electron spin-lattice relaxation time (T{sub 1e}) of TEMPO- and trityl-doped butanol samples at 2.5 T and temperatures between 0.95 K and 2.17 K was studied by pulsed nuclear magnetic resonance (NMR) using the nuclear-electron double resonance (NEDOR) method. This method is based on the idea to measure the NMR lineshift produced by the local field of paramagnetic impurities, whose polarization can be manipulated. This is of technical advantage as measurements can be performed under conditions typically used for the dynamic nuclear polarization (DNP) process - in our case 2.5 T and temperatures around 1 K - where a direct measurement on the electronic spins would be far more complicated to perform. As T{sub 1e} is a crucial parameter determining the overall efficiency of DNP, the effect of the radical type, its spin concentration, the temperature and the oxygen content on T{sub 1e} has been investigated. For radical concentrations as used in DNP (several 10{sup 19} spins/cm{sup 3}) the relaxation rate (T{sub 1e}{sup -1}) has shown a linear dependence on the paramagnetic electron concentration for both radicals investigated. Experiments with perdeuterated and ordinary butanol have given no indication for any influence of the host materials isotopes. The measured temperature dependence has shown an exponential characteristic. It is further observed that the oxygen content in the butanol samples has a considerable effect on the electron relaxation time and thus influences the nuclear relaxation time and polarization rate during the DNP. The experiments also show a variation in the NMR linewidth, leading to comparable time constants as determined by the lineshift. NEDOR measurements were also performed on irradiated, crystal grains of {sup 6}LiD. These samples exhibited a linewidth behavior similar to that of the cylindrically shaped butanol samples.

  6. Documentary Realism, Sampling Theory and Peircean Semiotics: electronic audiovisual signs (analog or digital as indexes of reality

    Directory of Open Access Journals (Sweden)

    Hélio Godoy

    2007-07-01

    Full Text Available This paper addresses Documentary Realism, focusing on thephysical phenomena of transduction that take place in analog and digital audiovisual systems, herein analyzed in the light of the Sampling Theory, within the framework of Shannon and Weaver’s Information Theory. Transduction is a process by which one type of energy is transformed into another, or by which information is transcodified. Within the scope of Documentary Realism, it cannotbe claimed that electronic audiovisual signs, because of their technical digital features lead to a rupture with reality. Rather, the digital documentary, based on electronic digital cinematography, is still an index of reality.

  7. Size-dependent ultrafast ionization dynamics of nanoscale samples in intense femtosecond x-ray free-electron-laser pulses.

    Science.gov (United States)

    Schorb, Sebastian; Rupp, Daniela; Swiggers, Michelle L; Coffee, Ryan N; Messerschmidt, Marc; Williams, Garth; Bozek, John D; Wada, Shin-Ichi; Kornilov, Oleg; Möller, Thomas; Bostedt, Christoph

    2012-06-08

    All matter exposed to intense femtosecond x-ray pulses from the Linac Coherent Light Source free-electron laser is strongly ionized on time scales competing with the inner-shell vacancy lifetimes. We show that for nanoscale objects the environment, i.e., nanoparticle size, is an important parameter for the time-dependent ionization dynamics. The Auger lifetimes of large Ar clusters are found to be increased compared to small clusters and isolated atoms, due to delocalization of the valence electrons in the x-ray-induced nanoplasma. As a consequence, large nanometer-sized samples absorb intense femtosecond x-ray pulses less efficiently than small ones.

  8. EVALUATION OF COMPUTER-CONTROLLED SCANNING ELECTRON MICROSCOPY APPLIED TO AN AMBIENT URBAN AEROSOL SAMPLE

    Science.gov (United States)

    Recent interest in monitoring and speciation of particulate matter has led to increased application of scanning electron microscopy (SEM) coupled with energy-dispersive x-ray analysis (EDX) to individual particle analysis. SEM/EDX provides information on the size, shape, co...

  9. Pseudomonas aeruginosa contamination of mouth swabs during production causing a major outbreak

    Directory of Open Access Journals (Sweden)

    Lassen Jørgen

    2007-03-01

    Full Text Available Abstract Background In 2002 we investigated an outbreak comprising 231 patients in Norway, caused by Pseudomonas aeruginosa and linked to the use of contaminated mouth swabs called Dent-O-Sept. Here we describe the extent of contamination of the swabs, and identify critical points in the production process that made the contamination possible, in order to prevent future outbreaks. Methods Environmental investigation with microbiological examination of production, ingredients and product, molecular typing of bacteria and a system audit of production. Results Of the 1565 swabs examined from 149 different production batches the outbreak strain of P. aeruginosa was detected in 76 swabs from 12 batches produced in 2001 and 2002. In total more than 250 swabs were contaminated with one or more microbial species. P. aeruginosa was detected from different spots along the production line. The audit revealed serious breeches of production regulations. Health care institutions reported non-proper use of the swabs and weaknesses in their purchasing systems. Conclusion Biofilm formation in the wet part of the production is the most plausible explanation for the continuous contamination of the swabs with P. aeruginosa over a period of at least 30 weeks. When not abiding to production regulations fatal consequences for the users may ensue. For the most vulnerable patient groups only documented quality-controlled, high-level disinfected products and items should be used in the oropharynx.

  10. A Comparison of Tissue versus Swab Culturing of Infected Diabetic Foot Wounds

    Directory of Open Access Journals (Sweden)

    Ying Huang

    2016-01-01

    Full Text Available Objective. To compare the efficacy of swabbing versus tissue biopsy for microbiological diagnosis of diabetic foot infection. Methods. This was a prospective trial. Fifty-six patients with diabetic foot infection were divided into the following 3 groups according to the PEDIS grading system: grade 2 (n=10, grade 3 (n=29, and grade 4 (n=17. Two specimens were collected from each wound for microbial culturing after debridement, including a superficial swab and a deep tissue punch biopsy specimen. Results. Swab culturing identified all of the microorganisms isolated from the corresponding deep tissue specimens in 9/10 of grade 2 wounds (90.0%, and this proportion decreased to 12/29 (41.4% and 7/17 (41.2% for grades 3 and 4 wounds, respectively (p=0.02. Moreover, the sensitivity for identifying Gram-negative bacteria, such as E. coli and Citrobacter, by swabbing was low (33.3%. In addition, some Gram-negative bacteria, such as Serratia and Ralstonia pickettii, were isolated from deep tissues but not from swabs. Conclusions. Swab culturing may be reliable for identification of pathogens in diabetic foot wounds classified as grade 2. However, it is advisable to culture deep tissue specimens for wounds of grade ≥3 because swab culturing is associated with a high risk of missing pathogens, especially Gram-negative bacteria.

  11. The Development of Indicator Cotton Swabs for the Detection of pH in Wounds

    Directory of Open Access Journals (Sweden)

    Cindy Schaude

    2017-06-01

    Full Text Available Indicator cotton swabs have been developed in order to enable faster, less expensive, and simpler information gathering of a wound status. Swabs are normally used for cleaning the wound, but here, they were covalently functionalized with indicator chemistry. Thus, they in principle enable simultaneous wound cleaning and wound pH detection. Using an indicator dye with a color change from yellow to red, combined with an inert dye of blue color, a traffic light color change from green to red is induced when pH increases. The indicator cotton swabs (ICSs show a color change from green (appropriate wound pH to red (elevated wound pH. This color change can be interpreted by the naked eye as well as by an optical color measurement device in order to obtain quantitative data based on the CIE L*a*b* color space. Two types of swabs have been developed—indicator cotton swabs ICS1 with a sensitive range from pH 5 to 7 and swabs ICS2 with a sensitive range from 6.5 to 8.5. The swabs are gamma-sterilized and the effect of sterilization on performance was found to be negligible. Furthermore, cytotoxicity testing shows cell viability and endotoxin levels to be within the allowable range.

  12. The Development of Indicator Cotton Swabs for the Detection of pH in Wounds.

    Science.gov (United States)

    Schaude, Cindy; Fröhlich, Eleonore; Meindl, Claudia; Attard, Jennifer; Binder, Barbara; Mohr, Gerhard J

    2017-06-12

    Indicator cotton swabs have been developed in order to enable faster, less expensive, and simpler information gathering of a wound status. Swabs are normally used for cleaning the wound, but here, they were covalently functionalized with indicator chemistry. Thus, they in principle enable simultaneous wound cleaning and wound pH detection. Using an indicator dye with a color change from yellow to red, combined with an inert dye of blue color, a traffic light color change from green to red is induced when pH increases. The indicator cotton swabs (ICSs) show a color change from green (appropriate wound pH) to red (elevated wound pH). This color change can be interpreted by the naked eye as well as by an optical color measurement device in order to obtain quantitative data based on the CIE L*a*b* color space. Two types of swabs have been developed-indicator cotton swabs ICS1 with a sensitive range from pH 5 to 7 and swabs ICS2 with a sensitive range from 6.5 to 8.5. The swabs are gamma-sterilized and the effect of sterilization on performance was found to be negligible. Furthermore, cytotoxicity testing shows cell viability and endotoxin levels to be within the allowable range.

  13. [Correlation between microbial growth in conjunctival swabs of corneal donors and contamination of organ culture media].

    Science.gov (United States)

    Li, S; Bischoff, M; Schirra, F; Langenbucher, A; Ong, M; Halfmann, A; Herrmann, M; Seitz, B

    2014-06-01

    The aim of the study was to determine the rate of contamination in conjunctival swabs from corneal donors by microbiological investigations and to correlate this with microbial contamination of the culture medium. Contamination of conjunctival swabs and culture media was analyzed retrospectively for the years 2009, 2010 and 2011 at the LIONS corneal bank of Saar-Lor-Lux Trier/Westpfalz at the Saarland University Medical Center. The total annual number of conjunctival swabs was 316 in 2009, 341 in 2010 and 381 in 2011. Conjunctival swabs were taken prior to 1.25% povidone-iodine application. After disinfection donor corneas were harvested by in situ corneoscleral disc excision in all cases. The correlation between positive conjunctival swabs and microbial contamination of the culture medium was analyzed. In every year examined the contamination rate of the culture medium was significantly higher in cases of contaminated conjunctival swabs (p culture medium was contaminated in 16.5%, 11.5% and 7.6% of the donated corneas with positive conjunctival swabs and in 7.2%, 1.9% and 0.6% in donated corneas with negative conjunctival swabs, respectively. A positive correlation was found between contamination of the culture medium and microbial colonization of the conjunctival swabs, Nevertheless, microbial colonization of the conjunctiva was high and contamination of the culture medium was relatively low. For the microbial contamination rate of the donated corneas in the medium, conjunctival disinfection time with iodine solution before explantation of the corneoscleral disc and the addition of antibiotics to the culture medium seem to play a protective role.

  14. Modeling of the interaction of an x-ray free-electron laser with large finite samples

    Science.gov (United States)

    Peyrusse, O.; André, J.-M.; Jonnard, P.; Gaudin, J.

    2017-10-01

    We describe a model for the study of the interaction of short x-ray free-electron laser (XFEL) pulses with large finite samples. Hydrodynamics is solved in one-dimensional planar geometry with consideration of the electron-ion energy exchange and of the possible elastoplastic behavior. From a time-dependent calculation of the complex refractive index and of the underlying atomic physics, XFEL energy deposition is modeled through a calculation of the radiation field in the material. In the case of hard x-ray irradiation, energetic electrons induced by the XFEL absorption can propagate and deposit their energy outside the interaction region. Simulations of the interaction of hard x-ray ultrashort pulses with solid materials Ru and Si at different grazing incidence angles are presented and discussed. The results obtained demonstrate the potential of this approach to predict damage dynamics for materials of interest for x-ray optics.

  15. Current Microbial Isolates from Wound Swab and Their Susceptibility Pattern in a Private Medical College Hospital in Dhaka city

    Directory of Open Access Journals (Sweden)

    Shahin Sultana

    2015-03-01

    Full Text Available Background: Wound infection is one of the major health problems that are caused and aggravated by the invasion of pathogenic organisms where empiric treatment is routine. Objective: To isolate and identify the bacteria causing wound infection and to determine the antimicrobial susceptibility pattern. Materials and method: A total of 263 wound swab and pus samples were collected during the period of January to December 2012 from Delta Medical College and Hospital, Dhaka, Bangladesh. Swabs from the wound were inoculated on appropriate media and cultured and the isolates were identified by standard procedures as needed. Antimicrobial susceptibility testing was performed by disk diffusion method according to ‘The Clinical Laboratory Standard Institute’ guidelines. Results: In this study 220 bacterial isolates were recovered from 263 samples showing an isolation rate of 83.65%. The predominant bacteria isolated from infected wounds were Staphylococcus aureus 89 (40.45% followed by Escherichia coli 62 (28.18%, Pseudomonas aeruginosa 34 (15.45%, Enterococci 18 (8.18%, Acinetobacter 5 (2.27%, Klebsiella 9 (4.09% and Proteus 3 (3.36%. Staphylococcus aureus was sensitive to linezolid (94.38%, fusidic acid (91.01%, vancomycin (87.64%, amikacin (74.15% and gentamicin (73.03%. Among the Gram negative isolates Escherichia coli was predominant and showed sensitivity to imipenem (93.54% amikacin (83.87% colistin (53.22% and piperacillin and tazobactum (53.22% and pseudomonas showed sensitivity to amikacin (73.52%, imipenem (70.58% and colistin (70.58%. Conclusion: Staphylococcus aureus was the most frequently isolated pathogen from wound swab and the antibiotic sensitivity pattern of various isolates help to assist the clinician in appropriate selection of empirical antibiotics against wound infection.

  16. Examination of mycological samples by means of the scanning electron microscope

    Directory of Open Access Journals (Sweden)

    M. Thibaut

    1973-04-01

    Full Text Available Three species of Siphomycetes: Rhizopus arhizus, Rhizopus equinus and Rhizopus nigricans, as well as a Septomycete: Emericella nidulans, have been examined by means of a scanning electron microscope. Among the difjerent Rhizopus, this technique showed differences in the appearance of the sporangia. In Emericella nidulans, scanning microscopy enábled one to ascertain that the "Hull cells" were completely hollow and also demonstrated the ornemented aspect of the ascospores.

  17. Investigation of Deuterium Implantation into Beryllium Sample by Electron Energy Loss Spectroscopy

    Science.gov (United States)

    Afanas’ev, V. P.; Gryazev, A. S.; Kaplya, P. S.; Köppen, M.; Ridzel, O. Yu; Subbotin, N. Yu; Hansen, P.

    2017-11-01

    Quantitative analysis of hydrogen isotopes in first wall as well as in construction materials of future fusion devices plays a crucial role to understand the evolution of those materials under operation conditions. A quantitative understanding of hydrogen in materials is also an important issue for storing energy as well as for fuel cells. A combination of Electron Energy Loss Spectroscopy (EELS) and Elastic Peak Electron Spectroscopy (EPES) is presented in this study to tackle these problems of modern material research for energy production and storage. Accurate inelastic scattering background subtraction is a key part of the presented quantitative measurement of the Be/D ratio. The differential inelastic scattering cross-section is determined by the fitting procedure. The fitting procedure is based on the iterative solution of the direct problem and minimization of the residual between computed and measured spectra. This study also takes into account the difference in electron energy loss laws for surface and bulk. The inelastic scattering cross-sections for different doses of deuterium ions in beryllium substrate (5.5·1021 m‑2 and 2.01·1022 m‑2) were defined in a two-layered model. The analysis is carried out for the EELS spectra. Relative concentration of D atoms is defined.

  18. 3D printing scanning electron microscopy sample holders: A quick and cost effective alternative for custom holder fabrication.

    Science.gov (United States)

    Meloni, Gabriel N; Bertotti, Mauro

    2017-01-01

    A simple and cost effective alternative for fabricating custom Scanning Electron Microscope (SEM) sample holders using 3D printers and conductive polylactic acid filament is presented. The flexibility of the 3D printing process allowed for the fabrication of sample holders with specific features that enable the high-resolution imaging of nanoelectrodes and nanopipettes. The precise value of the inner semi cone angle of the nanopipettes taper was extracted from the acquired images and used for calculating their radius using electrochemical methods. Because of the low electrical resistivity presented by the 3D printed holder, the imaging of non-conductive nanomaterials, such as alumina powder, was found to be possible. The fabrication time for each sample holder was under 30 minutes and the average cost was less than $0.50 per piece. Despite being quick and economical to fabricate, the sample holders were found to be sufficiently resistant, allowing for multiple uses of the same holder.

  19. 3D printing scanning electron microscopy sample holders: A quick and cost effective alternative for custom holder fabrication.

    Directory of Open Access Journals (Sweden)

    Gabriel N Meloni

    Full Text Available A simple and cost effective alternative for fabricating custom Scanning Electron Microscope (SEM sample holders using 3D printers and conductive polylactic acid filament is presented. The flexibility of the 3D printing process allowed for the fabrication of sample holders with specific features that enable the high-resolution imaging of nanoelectrodes and nanopipettes. The precise value of the inner semi cone angle of the nanopipettes taper was extracted from the acquired images and used for calculating their radius using electrochemical methods. Because of the low electrical resistivity presented by the 3D printed holder, the imaging of non-conductive nanomaterials, such as alumina powder, was found to be possible. The fabrication time for each sample holder was under 30 minutes and the average cost was less than $0.50 per piece. Despite being quick and economical to fabricate, the sample holders were found to be sufficiently resistant, allowing for multiple uses of the same holder.

  20. Note: Adhesive stamp electrodes using spider silk masks for electronic transport measurements of supra-micron sized samples

    Science.gov (United States)

    Steven, E.; Jobiliong, E.; Eugenio, P. M.; Brooks, J. S.

    2012-04-01

    A procedure for fabricating adhesive stamp electrodes based on gold coated adhesive tape used to measure electronic transport properties of supra-micron samples in the lateral range 10-100 μm and thickness >1 μm is described. The electrodes can be patterned with a ˜4 μm separation by metal deposition through a mask using Nephila clavipes spider dragline silk fibers. Ohmic contact is made by adhesive lamination of a sample onto the patterned electrodes. The performance of the electrodes with temperature and magnetic field is demonstrated for the quasi-one-dimensional organic conductor (TMTSF)2PF6 and single crystal graphite, respectively.

  1. A new fractionator principle with varying sampling fractions: exemplified by estimation of synapse number using electron microscopy

    DEFF Research Database (Denmark)

    Witgen, Brent Marvin; Grady, M. Sean; Nyengaard, Jens Randel

    2006-01-01

    incorporates a varying sampling fraction paradigm. The method allows for systematic random sampling from blocks of variable slab thickness, thereby eliminating the need for exhaustive serial sectioning through an entire containing space. This novel approach acknowledges the inaccuracy inherent in estimating......The quantification of ultrastructure has been permanently improved by the application of new stereological principles. Both precision and efficiency have been enhanced. Here we report for the first time a fractionator method that can be applied at the electron microscopy level. This new design...

  2. Direct PCR amplification of the HVSI region in mitochondrial DNA from buccal cell swabs

    Directory of Open Access Journals (Sweden)

    Kovačević-Grujičić Nataša

    2012-01-01

    Full Text Available Amplification of human mitochondrial DNA (mtDNA has been widely used in population genetics, human evolutionary and molecular anthropology studies. mtDNA hypervariable segments I and II (HVSI and HVSII were shown to be a suitable tool in genetic analyses due to the unique properties of mtDNA, such as the lack of recombination, maternal mode of inheritance, rapid evolutionary rate and high population-specific polymorphisms. Here we present a rapid and low-cost method for direct PCR amplification of a 330 bp fragment of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this method appropriate for the analysis of a large number of samples in a short period of time. Since the transportation of samples and fieldwork conditions can affect the quality of samples and subsequent DNA analysis, we tested the effects of long-term storage of buccal cell swabs on the suitability of such samples for direct PCR amplification. We efficiently amplified a 330 bp fragment of HVSI even after the long-term storage of buccal cells at room temperature, +4°C or at -20°C, for up to eight months. All examined PCR products were successfully sequenced, regardless of sample storage time and conditions. Our results suggest that the direct PCR amplification of the HVSI region from buccal cells is a method well suited for large-scale mtDNA population studies.[Acknowledgments. This work was supported by the Ministry of Education and Science of the Republic of Serbia (Grant no. III 47025.

  3. An evaluation of sampling methods for the detection of Escherichia coli and Salmonella on Turkey carcasses.

    Science.gov (United States)

    McEvoy, J M; Nde, C W; Sherwood, J S; Logue, C M

    2005-01-01

    The efficacy of rinse, excision, and swab methods for the microbiological analysis of prechill turkey carcasses was investigated. Aerobic plate counts from a 50-cm2 area of the breast sampled by excision and by swabbing were compared. Escherichia coli and Salmonella recoveries were determined from turkeys sampled by a carcass rinse (CR), a modified rinse with the carcass supported in a swing (MCR), a two-site swab of 50 cm2 at the back and thigh (2S), a one-site swab of 50 cm2 beneath the wing (1S), a whole-carcass swab of the inner and outer carcass surface (WS), and excision of 25 g of neck skin tissue (NE). The effect of diluent volume (25, 50, and 100 ml) on E. coli counts from swab samples was also assessed. The aerobic plate count from breast tissue sampled by excision was greater than that by swabbing (P diluent (P diluent, E. coli recoveries by the MCR, 2S, 1S, and WS methods were similar. For swabs stomached in 50 ml of diluent, Salmonella recoveries by the WS and MCR methods were higher than those by the 2S and 1S methods. Excision was more effective than swabbing for obtaining total bacterial counts from reduced turkey carcass areas. Whole-carcass sampling by rinsing or swabbing is necessary for optimum Salmonella recovery. Sampling a reduced area of the carcass is sufficient for E. coli analysis.

  4. Composition of plastics from waste electrical and electronic equipment (WEEE) by direct sampling.

    Science.gov (United States)

    Martinho, Graça; Pires, Ana; Saraiva, Luanha; Ribeiro, Rita

    2012-06-01

    This paper describes a direct analysis study carried out in a recycling unit for waste electrical and electronic equipment (WEEE) in Portugal to characterize the plastic constituents of WEEE. Approximately 3400 items, including cooling appliances, small WEEE, printers, copying equipment, central processing units, cathode ray tube (CRT) monitors and CRT televisions were characterized, with the analysis finding around 6000 kg of plastics with several polymer types. The most common polymers are polystyrene, acrylonitrile-butadiene-styrene, polycarbonate blends, high-impact polystyrene and polypropylene. Additives to darken color are common contaminants in these plastics when used in CRT televisions and small WEEE. These additives can make plastic identification difficult, along with missing polymer identification and flame retardant identification marks. These drawbacks contribute to the inefficiency of manual dismantling of WEEE, which is the typical recycling process in Portugal. The information found here can be used to set a baseline for the plastics recycling industry and provide information for ecodesign in electrical and electronic equipment production. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Self-collected vaginal swabs for HPV screening: An exploratory study of rural Black Mississippi women

    Directory of Open Access Journals (Sweden)

    Richard A. Crosby

    2017-09-01

    Conclusions: Black rural women from the deep-south are generally comfortable self-collecting cervico-vaginal swabs for HPV testing. Given that nearly 30% tested positive for oncogenic HPV, and that fatalism as well a lack of trust in doctors predicted prevalence, a reasonable screening alternative to Pap testing may be community-based testing for HPV using self-collected vaginal swabs.

  6. Mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swab specimens in Brazil

    OpenAIRE

    dos Santos, Danielle Caldeira Martins; da Costa, Thaina Miranda; Rabello, Renata Fernandes; Alves, F?bio Aguiar; de Mondino, Silvia Susana Bona

    2015-01-01

    The isolation of mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swabs is reported. Among the 59 isolates, 9 (15%) isolates were mannitol-negative; all of these isolates were categorized as staphylococcal cassette chromosome mec (SCCmec) type IVa. This report emphasizes that mannitol fermentation on mannitol salt agar should not be used as the sole criterion when screening nasal swab specimens for S. aureus.

  7. Mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swab specimens in Brazil

    Directory of Open Access Journals (Sweden)

    Danielle Caldeira Martins dos Santos

    2015-06-01

    Full Text Available The isolation of mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swabs is reported. Among the 59 isolates, 9 (15% isolates were mannitol-negative; all of these isolates were categorized as staphylococcal cassette chromosome mec (SCCmec type IVa. This report emphasizes that mannitol fermentation on mannitol salt agar should not be used as the sole criterion when screening nasal swab specimens for S. aureus.

  8. Mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swab specimens in Brazil.

    Science.gov (United States)

    dos Santos, Danielle Caldeira Martins; da Costa, Thaina Miranda; Rabello, Renata Fernandes; Alves, Fábio Aguiar; de Mondino, Silvia Susana Bona

    2015-06-01

    The isolation of mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swabs is reported. Among the 59 isolates, 9 (15%) isolates were mannitol-negative; all of these isolates were categorized as staphylococcal cassette chromosome mec (SCCmec) type IVa. This report emphasizes that mannitol fermentation on mannitol salt agar should not be used as the sole criterion when screening nasal swab specimens for S. aureus.

  9. "DETECTION OF BACTERIAL, METHICILLIN RESISTANCE, AND β-LACTAMASE GENES FOUND IN WOUND SWABS BY MULTIPLEX POLYMERASE CHAIN REACTION"

    Directory of Open Access Journals (Sweden)

    S. Sadeghian

    2004-05-01

    Full Text Available Coagulase-positive and coagulase negative, methicillin-resistant staphylococci are major causes of serious nosocomial infections and it is very important to have a reliable test to detect these bacteria. A multiplex polymerase chain reaction (mPCR was used on 100 clinical samples for simultaneous amplification of the universal bacterial, mec-A encoding the penicillin binding protein 2a, which is associated with staphylococcal methicillin resistance and TEM-1 encoding the β-lactamase, which accounts for the majority of all cases of the plasmid β-lactamase resistance worldwide. Out of 100 wound swabs tested, 99% with universal primers, 26% with TEM-1 primers and 6% with mec-A primers were positive. Dot blot Digoxigenin hybridization on the 30 samples was carried out to confirm identified bacteria with specific bacterial probes. Out of 100 wound swabs, 38% were positive with Staphylococcus aureus probe, 23% were positive with enteric bacteria probe, 7% were positive with Streptococcus agalactia probe and 1% were positive with Haemophilus influenza probe. The mPCR method used in this study, was designed to be incorporated into the workflow of the clinical microbiology laboratory and allows for the identification of intrinsic resistance in a timely and reliable manner.

  10. Examination of mycological samples by means of the scanning electron microscope

    Directory of Open Access Journals (Sweden)

    M. Thibaut

    1973-04-01

    Full Text Available Three species of Siphomycetes: Rhizopus arhizus, Rhizopus equinus and Rhizopus nigricans, as well as a Septomycete: Emericella nidulans, have been examined by means of a scanning electron microscope. Among the difjerent Rhizopus, this technique showed differences in the appearance of the sporangia. In Emericella nidulans, scanning microscopy enábled one to ascertain that the "Hull cells" were completely hollow and also demonstrated the ornemented aspect of the ascospores.Três espécies de Sifomicetas: Rhizopus arhizus, Rhizopus equinus, Rhizopus nigricans e um Septomiceta: Emericella nidulans foram examinados em microscopia de exploração. Esta técnica mostrou detalhes não evidenciáveis ao poder de resolução do microscópio óptico, demonstrando ser útil para o diagnóstico em micologia.

  11. The Use of Multiplex PCR to Determine the Prevalence of Enterotoxigenic Staphylococcus aureus Isolated from Raw Milk, Feta Cheese, and Hand Swabs.

    Science.gov (United States)

    Zeinhom, Mohamed M A; Abdel-Latef, Gihan K; Jordan, Kieran

    2015-12-01

    Staphylococcus aureus (S. aureus) can cause mastitis in cattle and, therefore, can be present in milk. This study was undertaken to determine the prevalence of coagulase positive S. aureus and its enterotoxin genes sea, seb, and sec in isolates recovered from raw milk, feta cheese, and human hand swabs of milk and cheese handlers in Beni-Suef province, Egypt. A total of 100 samples of raw milk and 50 samples of pasteurized-milk feta cheese were collected. In addition, 50 hand swabs from milk handlers and 25 hand swabs from cheese handlers were examined for the presence of coagulase positive S. aureus. The isolates were characterized by multiplex PCR for detection of sea, seb, and sec genes, and for resistance to 5 classes of commonly used antibiotics. Twelve (12/100), 12 (6/50), and 17% (13/75) of milk, cheese, and hand swab samples, respectively, were positive for coagulase positive S. aureus. One isolate was obtained from each positive sample (31 isolates), and none contained genes for SEA or SEC production. Twenty-five percent, 33%, and 31%, respectively, of the isolates contained the genes for SEB, resulting in 3%, 4%, and 5% of samples being positive for toxin producing coagulase positive S. aureus, respectively. At least one isolate was resistant to each of the antibiotics tested. Despite the low potential for SEB production shown, preventative measures, such as maintenance of the cold-chain and good hygienic practices should be implemented to further reduce the potential risk to public health from SEB, and to reduce the spread of antimicrobial resistance. © 2015 Institute of Food Technologists®

  12. ``Low-cost Electronic nose evaluated on Thai-herb of Northern-Thailand samples using multivariate analysis methods''

    Science.gov (United States)

    na ayudhaya, Paisarn Daungjak; Klinbumrung, Arrak; Jaroensutasinee, Krisanadej; Pratontep, Sirapat; Kerdcharoen, Teerakiat

    2009-05-01

    In case of species of natural and aromatic plant originated from the northern Thailand, sensory characteristics, especially odours, have unique identifiers of herbs. The instruments sensory analysis have performed by several of differential of sensing, so call `electronic nose', to be a significantly and rapidly for chemometrics. The signal responses of the low cost electronic nose were evaluated by principal component analysis (PCA). The aims of this paper evaluated various of Thai-herbs grown in Northern of Thailand as data preprocessing tools of the Low-cost electronic nose (enNU-PYO1). The essential oil groups of Thai herbs such as Garlic, Lemongrass, Shallot (potato onion), Onion, Zanthoxylum limonella (Dennst.) Alston (Thai name is Makaen), and Kaffir lime leaf were compared volatilized from selected fresh herbs. Principal component analysis of the original sensor responses did clearly distinguish either all samples. In all cases more than 97% for cross-validated group were classified correctly. The results demonstrated that it was possible to develop in a model to construct a low-cost electronic nose to provide measurement of odoriferous herbs.

  13. Monte Carlo Simulation of Characteristic Secondary Fluorescence in Electron Probe Microanalysis of Homogeneous Samples Using the Splitting Technique.

    Science.gov (United States)

    Petaccia, Mauricio; Segui, Silvina; Castellano, Gustavo

    2015-06-01

    Electron probe microanalysis (EPMA) is based on the comparison of characteristic intensities induced by monoenergetic electrons. When the electron beam ionizes inner atomic shells and these ionizations cause the emission of characteristic X-rays, secondary fluorescence can occur, originating from ionizations induced by X-ray photons produced by the primary electron interactions. As detectors are unable to distinguish the origin of these characteristic X-rays, Monte Carlo simulation of radiation transport becomes a determinant tool in the study of this fluorescence enhancement. In this work, characteristic secondary fluorescence enhancement in EPMA has been studied by using the splitting routines offered by PENELOPE 2008 as a variance reduction alternative. This approach is controlled by a single parameter NSPLIT, which represents the desired number of X-ray photon replicas. The dependence of the uncertainties associated with secondary intensities on NSPLIT was studied as a function of the accelerating voltage and the sample composition in a simple binary alloy in which this effect becomes relevant. The achieved efficiencies for the simulated secondary intensities bear a remarkable improvement when increasing the NSPLIT parameter; although in most cases an NSPLIT value of 100 is sufficient, some less likely enhancements may require stronger splitting in order to increase the efficiency associated with the simulation of secondary intensities.

  14. Experimental Design for a Macrofoam Swab Study Relating the Recovery Efficiency and False Negative Rate to Low Concentrations of Two Bacillus anthracis Surrogates on Four Surface Materials

    Energy Technology Data Exchange (ETDEWEB)

    Piepel, Gregory F.; Hutchison, Janine R.

    2014-04-16

    This report describes the experimental design for a laboratory study to quantify the recovery efficiencies and false negative rates of a validated, macrofoam swab sampling method for low concentrations of Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus (BG) spores on four surface materials (stainless steel, glass, vinyl tile, plastic light cover panel). Two analytical methods (plating/counting and polymerase chain reaction) will be used. Only one previous study has investigated false negative as a function of affecting test factors. The surrogates BAS and BG have not been tested together in the same study previously. Hence, this study will provide for completing gaps in the available information on the performance of macrofoam swab sampling at low concentrations.

  15. Experimental Design for a Macrofoam-Swab Study Relating the Recovery Efficiency and False Negative Rate to Low Concentrations of Two Bacillus anthracis Surrogates on Four Surface Materials

    Energy Technology Data Exchange (ETDEWEB)

    Piepel, Gregory F. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2014-12-05

    This report describes the experimental design for a laboratory study to quantify the recovery efficiencies and false negative rates of a validated, macrofoam-swab sampling method for low concentrations of Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus (BG) spores on four surface materials (stainless steel, glass, vinyl tile, plastic light cover panel). Two analytical methods (culture and polymerase chain reaction) will be used. Only one previous study has investigated how the false negative rate depends on test factors. The surrogates BAS and BG have not been tested together in the same study previously. Hence, this study will provide for completing gaps in the available information on the performance of macrofoam-swab sampling at low concentrations.

  16. MRT letter: localization of endogenous hydrogen peroxide by modified processes of sample preparation for transmission electron microscope in Escherichia coli.

    Science.gov (United States)

    Li, Xin; Hu, Rongliu; Zhu, Wenxue; Fan, Jinling; Pang, Xinyue; Wang, Na; Wang, Liping; Yang, Lipeng; Zhao, Chunyan; He, Chenyang

    2013-02-01

    The bacterial endogenous hydrogen peroxide (H(2)O(2)) was detected cytochemically by its reaction with cerium chloride (CeCl(3)) to produce electron-dense deposits of cerium perhydroxides. The sequence of fixation and CeCl(3) staining of H(2)O(2) in the processing of transmission electron microscope (TEM) sample preparation is crucial to the localization of endogenous H(2)O(2) in Escherichia coli. In this study, results confirmed that the process that fixation simultaneously with CeCl(3) staining provided optimum effects for H(2)O(2) localization in E. coli. The modified process of TEM provides very efficient protection for H(2)O(2) localization and more accurate quantization for the H(2)O(2) accumulation in bacterial cells. Copyright © 2012 Wiley Periodicals, Inc.

  17. Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru.

    Science.gov (United States)

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Reyes-Batlle, María; González, Ana C; Guerra, Humberto; Gotuzzo, Eduardo; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Free Living Amoebae (FLA) of Acanthamoeba genus are widely distributed in the environment and can be found in the air, soil and water; and have also been isolated from air-conditioning units. In humans, they are causative agents of a sight-threating infection of the cornea, Acanthamoeba keratitis (AK) and a fatal infection of the central nervous system known as Granulomatous Amoebic Encephalitis (GAE). In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in nasal swabs from individuals in two regions of Peru. Identification of isolates was based on cyst morphology and PCR-sequencing of the Diagnostic Fragment 3 to identify strains at the genotype level. The pathogenic potential of the isolates was also assayed using temperature and osmotolerance assays and extracellular proteases zymograms. The obtained results revealed that all isolated strains exhibited pathogenic potential. After sequencing the highly variable DF3 (Diagnostic Fragment 3) region in the 18S rRNA gene as previously described, genotype T4 was found to be the most common one in the samples included in this study but also genotype T15 was identified. To the best of our knowledge, this is the first study on the characterization of Acanthamoeba strains at the genotype level and the first report of genotype T4 and T15 in Peru. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Chicken parvovirus viral loads in cloacal swabs from malabsorption syndrome-affected and healthy broilers.

    Science.gov (United States)

    Finkler, Fabrine; de Lima, Diane Alves; Cerva, Cristine; Cibulski, Samuel Paulo; Teixeira, Thais Fumaco; Dos Santos, Helton Fernandes; de Almeida, Laura Lopes; Roehe, Paulo Michel; Franco, Ana Cláudia

    2016-12-01

    Chicken parvovirus (ChPV) has been associated with malabsorption syndrome (MAS) in broilers. However, the participation of this virus in such syndrome is unclear, since it may be detected in diseased and healthy chickens. In the course of these studies, it was argued whether ChPV genome loads might be correlated to the occurrence of MAS. To check such a hypothesis, a SYBR green-based quantitative polymerase chain reaction was developed to detect and quantify ChPV genomes. Cloacal swabs from 68 broilers with MAS and 59 from healthy animals were collected from different poultry farms. Genomes of ChPV were detected in all samples, regardless of their health status. However, viral genome loads in MAS-affected broilers were significantly higher (1 × 10 5 genome copies per 100 ng DNA) than in healthy animals (1.3 × 10 3 GC/100 ng DNA). These findings indicate that there is an association between high ChPV genome loads and the occurrence of MAS in broilers.

  19. Sample Injector Fabrication and Delivery Method Development for Serial Crystallography using Synchrotrons and X-ray Free Electron Lasers

    Science.gov (United States)

    Nelson, Garrett Charles

    Sample delivery is an essential component in biological imaging using serial diffraction from X-ray Free Electron Lasers (XFEL) and synchrotrons. Recent developments have made possible the near-atomic resolution structure determination of several important proteins, including one G protein-coupled receptor (GPCR) drug target, whose structure could not easily have been determined otherwise (Appendix A). In this thesis I describe new sample delivery developments that are paramount to advancing this field beyond what has been accomplished to date. Soft Lithography was used to implement sample conservation in the Gas Dynamic Virtual Nozzle (GDVN). A PDMS/glass composite microfluidic injector was created and given the capability of millisecond fluidic switching of a GDVN liquid jet within the divergent section of a 2D Laval-like GDVN nozzle, providing a means of collecting sample between the pulses of current XFELs. An oil/water droplet immersion jet was prototyped that suspends small sample droplets within an oil jet such that the sample droplet frequency may match the XFEL pulse repetition rate. A similar device was designed to use gas bubbles for synchronized "on/off" jet behavior and for active micromixing. 3D printing based on 2-Photon Polymerization (2PP) was used to directly fabricate reproducible GDVN injectors at high resolution, introducing the possibility of systematic nozzle research and highly complex GDVN injectors. Viscous sample delivery using the "LCP injector" was improved with a method for dealing with poorly extruding sample mediums when using full beam transmission from the Linac Coherent Light Source (LCLS), and a new viscous crystal-carrying medium was characterized for use in both vacuum and atmospheric environments: high molecular weight Polyethylene Glycol.

  20. Evaluation of Presto(plus) assay and LightMix kit Trichomonas vaginalis assay for detection of Trichomonas vaginalis in dry vaginal swabs.

    Science.gov (United States)

    de Waaij, Dewi J; Ouburg, Sander; Dubbink, Jan Henk; Peters, Remco P H; Morré, Servaas A

    2016-08-01

    This is an evaluation study of the Presto(plus) Assay for T. vaginalis by comparing to the TIB MOLBIOL LightMix Kit Trichomonas vaginalis Assay using 615 dry collected vaginal and rectal swabs. Discordant samples were analyzed by the Qiagen® Microbial DNA qPCR for TV Assay. Both assays showed comparable performances (McNemar p>0.05). Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Etiologic predictive value of a rapid immunoassay for the detection of group A Streptococcus antigen from throat swabs in patients presenting with a sore throat.

    Science.gov (United States)

    Orda, Ulrich; Gunnarsson, Ronny; Orda, Sabine; Fitzgerald, Mark; Rofe, Geoff; Dargan, Anna

    2016-04-01

    Clinical reasoning utilizing certain symptoms and scores has not proven to be a reliable decision-making tool to determine whether or not to suspect a group A Streptococcus (GAS) infection in the patient presenting with a sore throat. Culture as the so-called 'gold standard' is impracticable because it takes 1 to 2 days (and even longer in remote locations) for a result, and thus treatment decisions will be made without the result available. Rapid diagnostic antigen tests have demonstrated sufficient sensitivities and specificities in detecting GAS antigens to identify GAS throat infections. Throat swab samples were collected from patients attending the Mount Isa Hospital emergency department for a sore throat; these samples were compared to swab samples collected from healthy controls who did not have a sore throat. Both groups were aged 3-15 years. All swab samples were analyzed with a point-of-care test (Alere Test Pack +Plus with OBC Strep A). The etiologic predictive value (EPV) of the throat swab was calculated. The 95% confidence interval for positive EPV was 88-100% and for negative EPV was 97-99%, depending on assumptions made. This study demonstrates that the point-of-care test Alere Test Pack +Plus Strep A has a high positive predictive value and is able to rule in GAS infection as long as the proportion of carriers is low. Also the negative predictive value for ruling out GAS as the etiologic agent is very high irrespective of the carrier rate. Hence, this test is always useful to rule out GAS infection. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Apparatus for Sampling Surface Contamination

    Science.gov (United States)

    Wells, Mark

    2008-01-01

    An apparatus denoted a swab device has been developed as a convenient means of acquiring samples of contaminants from surfaces and suspending the samples in liquids. (Thereafter, the liquids can be dispensed, in controlled volumes, into scientific instruments for analysis of the contaminants.) The swab device is designed so as not to introduce additional contamination and to facilitate, simplify, and systematize the dispensing of controlled volumes of liquid into analytical instruments. The swab device is a single apparatus into which are combined all the equipment and materials needed for sampling surface contamination. The swab device contains disposable components stacked together on a nondisposable dispensing head. One of the disposable components is a supply cartridge holding a sufficient volume of liquid for one complete set of samples. (The liquid could be clean water or another suitable solvent, depending on the application.) This supply of liquid is sealed by Luer valves. At the beginning of a sampling process, the user tears open a sealed bag containing the supply cartridge. A tip on the nondisposable dispensing head is engaged with a Luer valve on one end of the supply cartridge and rotated, locking the supply cartridge on the dispensing head and opening the valve. The swab tip includes a fabric swab that is wiped across the surface of interest to acquire a sample. A sealed bag containing a disposable dispensing tip is then opened, and the swab tip is pushed into the dispensing tip until seated. The dispensing head contains a piston that passes through a spring-loaded lip seal. The air volume displaced by this piston forces the liquid out of the supply cartridge, over the swab, and into the dispensing tip. The piston is manually cycled to enforce oscillation of the air volume and thereby to cause water to flow to wash contaminants from the swab and cause the resulting liquid suspension of contaminants to flow into the dispensing tip. After several cycles

  3. Detection of Bordetella avium by TaqMan real-time PCR in tracheal swabs from wildlife birds.

    Science.gov (United States)

    Stenzel, T; Pestka, D; Tykałowski, B; Śmiałek, M; Koncicki, A; Bancerz-Kisiel, A

    2017-03-28

    Bordetella avium, the causing agent of bordetellosis, a highly contagious infection of the respiratory tract in young poultry, causes significant losses in poultry farming throughout the world. Wildlife birds can be a reservoir of various pathogens that infect farm animals. For this reason the studies were conducted to estimate the prevalence of Bordetella avium in wildlife birds in Poland. Tracheal swab samples were collected from 650 birds representing 27 species. The bacterial DNA was isolated directly from the swabs and screened for Bordetella avium by TaqMan real-time PCR. The assay specificity was evaluated by testing DNA isolated from 8 other bacteria that can be present in avian respiratory tract, and there was no amplification from non-Bordetella avium agents. Test sensitivity was determined by preparing standard tenfold serial dilutions of DNA isolated from positive control. The assay revealed to be sensitive, with detection limit of approximately 4.07x10^2 copies of Bordetella avium DNA. The genetic material of Bordetella avium was found in 54.54% of common pheasants, in 9.09% of Eurasian coots, in 3.22% of black-headed gulls and in 2.77% of mallard ducks. The results of this study point to low prevalence of Bordetella avium infections in wildlife birds. The results also show that described molecular assay proved to be suitable for the rapid diagnosis of bordetellosis in the routine diagnostic laboratory.

  4. Drop-on-demand sample delivery for studying biocatalysts in action at X-ray free-electron lasers.

    Science.gov (United States)

    Fuller, Franklin D; Gul, Sheraz; Chatterjee, Ruchira; Burgie, E Sethe; Young, Iris D; Lebrette, Hugo; Srinivas, Vivek; Brewster, Aaron S; Michels-Clark, Tara; Clinger, Jonathan A; Andi, Babak; Ibrahim, Mohamed; Pastor, Ernest; de Lichtenberg, Casper; Hussein, Rana; Pollock, Christopher J; Zhang, Miao; Stan, Claudiu A; Kroll, Thomas; Fransson, Thomas; Weninger, Clemens; Kubin, Markus; Aller, Pierre; Lassalle, Louise; Bräuer, Philipp; Miller, Mitchell D; Amin, Muhamed; Koroidov, Sergey; Roessler, Christian G; Allaire, Marc; Sierra, Raymond G; Docker, Peter T; Glownia, James M; Nelson, Silke; Koglin, Jason E; Zhu, Diling; Chollet, Matthieu; Song, Sanghoon; Lemke, Henrik; Liang, Mengning; Sokaras, Dimosthenis; Alonso-Mori, Roberto; Zouni, Athina; Messinger, Johannes; Bergmann, Uwe; Boal, Amie K; Bollinger, J Martin; Krebs, Carsten; Högbom, Martin; Phillips, George N; Vierstra, Richard D; Sauter, Nicholas K; Orville, Allen M; Kern, Jan; Yachandra, Vittal K; Yano, Junko

    2017-04-01

    X-ray crystallography at X-ray free-electron laser sources is a powerful method for studying macromolecules at biologically relevant temperatures. Moreover, when combined with complementary techniques like X-ray emission spectroscopy, both global structures and chemical properties of metalloenzymes can be obtained concurrently, providing insights into the interplay between the protein structure and dynamics and the chemistry at an active site. The implementation of such a multimodal approach can be compromised by conflicting requirements to optimize each individual method. In particular, the method used for sample delivery greatly affects the data quality. We present here a robust way of delivering controlled sample amounts on demand using acoustic droplet ejection coupled with a conveyor belt drive that is optimized for crystallography and spectroscopy measurements of photochemical and chemical reactions over a wide range of time scales. Studies with photosystem II, the phytochrome photoreceptor, and ribonucleotide reductase R2 illustrate the power and versatility of this method.

  5. Campylobacter sp in eggs from cloacal swab positive breeder hens Campylobacter sp em ovos provenientes de matrizes pesadas com swab cloacal positivo

    Directory of Open Access Journals (Sweden)

    Belchiolina Beatriz Fonseca

    2006-12-01

    Full Text Available Campylobacter sp is a microaerophilic, thermotolerating Gram negative bacterium, known to be one of the main causes of food-borne human infections. Among the foods that carry these microorganisms, the chicken is outstanding. In Brazil, a large chicken exporting country, few researches are conducted about their prevalence in breeder hens and the transmission through eggs. The aim of this research was to verify the presence of Campylobacter sp in the shells and within the eggs from positive cloacal swab breeder hens. Microbiological analyses were made on cloacal swabs of 140 weighed breeder hens. The positive breeder hens were set aside and in a total of 244 of their eggs, Campylobacter sp was present in macerated shells and yolk contents during 7 weeks. Out of the 140 researched breeder hens, 25 (17.8% were positive from cloacal swabs, however the eggs were not positive. The physiological characteristics of the birds, their eggs and Campylobacter sp favor the bacterium entering and surviving in the eggs, but in this study, no positive result was found in macerated shells or in the yolks, indicating that vertical transmission is probably an unusual event.Campylobacter sp é reconhecida como uma das principais causas de gastrenterite humana de origem alimentar. Dentre os alimentos veiculadores desses microrganismos, a carne de frango tem sido a mais implicada. Os estudos existentes sobre a transmissão vertical da Campylobacter são escassos e não conclusivos. O objetivo desse estudo foi verificar a presença de Campylobacter sp na casca e interior de ovos de matrizes positivas em swabs cloacais e a possibilidade de transmissão vertical. Foram analisados swabs cloacais de 140 matrizes pesadas e seus ovos colhidos para análise durante 7 semanas consecutivas. Dos 244 ovos colhidos, 129 foram fumigados e 115 analisados sem tratamento. Foram analisados o macerado da casca e a gema. Das 140 matrizes pesquisadas, 25 (17,8% foram positivas em swabs

  6. Self-collected vaginal swabs for the quantitative real-time polymerase chain reaction assay of Atopobium vaginae and Gardnerella vaginalis and the diagnosis of bacterial vaginosis.

    Science.gov (United States)

    Menard, J-P; Fenollar, F; Raoult, D; Boubli, L; Bretelle, F

    2012-04-01

    The aim of this study was to assess the feasibility of using self-collected vaginal specimens for the quantitative real-time polymerase chain reaction (qPCR) assays of bacterial vaginosis (BV)-associated bacteria versus practitioner-collected swabs. A cross-sectional study included 190 pregnant women enrolled before 20 weeks' gestation from September 2008 to November 2009. Self- and practitioner-collected swabs were taken during the same prenatal visit for each woman, qPCR assays performed for each, and the results compared. The quantification of the human albumin gene was used as an internal control to ensure sampling quality and accurate comparisons. The level of agreement of the qPCR assays for each microorganism was calculated with the Spearman product moment correlation coefficient and the kappa statistic. In all, 370 vaginal samples (185 self- and 185 practitioner-collected swabs) had a narrow range of values for the number of albumin gene copies and a significant correlation coefficient (Spearman's rho = 0.532; p Gardnerella vaginalis; p vaginalis (≥10(9) copies/mL; kappa value = 0.903; p vaginalis, and A. vaginae.

  7. Comparison of dacron and nylon-flocked self-collected vaginal swabs and urine for the detection of Trichomonas vaginalis using analyte-specific reagents in a transcription-mediated amplification assay.

    Science.gov (United States)

    Jang, Dan; Gilchrist, Jodi; Portillo, Eder; Smieja, Marek; Toor, Ramandeep; Chernesky, Max

    2012-04-01

    To compare self-collected vaginal swab (SCVS) types and first-catch urine (FCU) to diagnose Trichomonas vaginalis using analyte-specific reagents designed to be used in a transcription-mediated amplification assay. A total of 241 women (group A) collected a FCU and a SCVS using a dacron swab (APTIMA collection kit). A second group of 289 women (group B) collected two SCVS using one dacron swab and one nylon-flocked swab. Of 75 young women (street youth) determined to be infected with T vaginalis only seven reported symptoms of vaginal discharge or irritation. Using a cutoff of 50,000 relative light units, the sensitivity and specificity was 97.2% and 97.6%, respectively for dacron SCVS compared with 41.7% and 100% for FCU in group A; 92.3% and 98.8% for dacron SCVS and 92.3% and 99.2% for flocked-nylon SCVS in group B. The assay tested 96 samples in 6 h. Dacron and nylon-flocked SCVS performed equally well and significantly better than FCU using analyte-specific reagents in the APTIMA transcription-mediated amplification assay. Either swab type could be used for self-collection.

  8. A miniaturized and integrated gel post platform for multiparameter PCR detection of herpes simplex viruses from raw genital swabs.

    Science.gov (United States)

    Manage, Dammika P; Lauzon, Jana; Atrazhev, Alexey; Morrissey, Yuen C; Edwards, Ann L; Stickel, Alexander J; Crabtree, H John; Pabbaraju, Kanti; Zahariadis, George; Yanow, Stephanie K; Pilarski, Linda M

    2012-05-07

    Herpes simplex virus (HSV) is one of the most prevalent viruses, with acute and recurrent infections in humans. The current gold standard for the diagnosis of HSV is viral culture which takes 2-14 days and has low sensitivity. In contrast, DNA amplification by polymerase chain reaction (PCR) can be performed within 1-2 h. We here describe a multiparameter PCR assay to simultaneously detect HSV-1 and HSV-2 DNA templates, together with integrated positive and negative controls, with product detection by melting curve analysis (MCA), in an array of semi-solid polyacrylamide gel posts. Each gel post is 0.67 μL in volume, and polymerized with all the components required for PCR. Both PCR and MCA can currently be performed in one hour and 20 min. Unprocessed genital swabs collected in universal transport medium were directly added to the reagents before or after polymerization, diffusing from atop the gel posts. The gel post platform detects HSV templates in as little as 2.5 nL of raw sample. In this study, 45 genital swab specimens were tested blindly as a preliminary validation of this platform. The concordance of PCR on gel posts with conventional PCR was 91%. The primer sequestration method introduced here (wherein different primers are placed in different sets of posts) enables the simultaneous detection of multiple pathogens for the same sample, together with positive and negative controls, on a single chip. This platform accepts unprocessed samples and is readily adaptable to detection of multiple different pathogens or biomarkers for point-of-care diagnostics.

  9. Evaluation of Liquid-Based Swab Transport Systems against the New Approved CLSI M40-A2 Standard.

    Science.gov (United States)

    Gizzie, Nina; Adukwu, Emmanuel

    2016-04-01

    Following revised information pertaining to newer swab types and testing protocols in the new CLSI M40-A2 standard, we evaluated three liquid swab transport systems for the recovery of aerobic, anaerobic, and fastidious organisms at room temperature and at 4°C. All tested liquid swab transport systems were fully compliant with the M40-A2 standard, with acceptable performance at both temperatures after the full specified holding period, using both qualitative (roll-plate) and quantitative (swab elution) methods. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  10. Sample preparation of biological macromolecular assemblies for the determination of high-resolution structures by cryo-electron microscopy.

    Science.gov (United States)

    Stark, Holger; Chari, Ashwin

    2016-02-01

    Single particle cryo-EM has recently developed into a powerful tool to determine the 3D structure of macromolecular complexes at near-atomic resolution, which allows structural biologists to build atomic models of proteins. All technical aspects of cryo-EM technology have been considerably improved over the last two decades, including electron microscopic hardware, image processing software and the ever growing speed of computers. This leads to a more widespread use of the technique, and it can be anticipated that further automation of electron microscopes and image processing tools will soon fully shift the focus away from the technological aspects, onto biological questions that can be answered. In single particle cryo-EM, no crystals of a macromolecule are required. In contrast to X-ray crystallography, this significantly facilitates structure determination by cryo-EM. Nevertheless, a relatively high level of biochemical control is still essential to obtain high-resolution structures by cryo-EM, and it can be anticipated that the success of the cryo-EM technology goes hand in hand with further developments of sample purification and preparation techniques. This will allow routine high-resolution structure determination of the many macromolecular complexes of the cell that until now represent evasive targets for X-ray crystallographers. Here we discuss the various biochemical tools that are currently available and the existing sample purification and preparation techniques for cryo-EM grid preparation that are needed to obtain high-resolution images for structure determination. © The Author 2015. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Determination of pentachlorophenol in wood samples using liquid chromatography with UV absorbance, amperometric and electron-capture detection.

    Science.gov (United States)

    Goewie, C E; Berkhof, R J; Maris, F A; Treskes, M; Brinkman, U A

    1986-01-01

    The potential of reversed-phase liquid chromatography (RPLC) with UV and amperometric detection (AD), and of normal-phase liquid chromatography (NPLC) with UV and electron-capture detection (ECD) for the determination of pentachlorophenol (PCP) in wood samples has been studied. When PCP concentrations of at least 1-5 ppm have to be determined, RPLC-UV and RPLC-AD on C18-modified silica are useful techniques, provided a two- or three-step sample-preparation step is used. NPLC-UV on bare silica columns does not offer any advantage over RPLC-UV. NPLC-ECD on bare silica and with an acidified toluene-hexane mixture as eluent offers good selectivity and sensitivity, as well as satisfactory linearity and reproducibility for the determination of PCP in wood samples down to low ppb levels. Use of the two-step clean-up procedure is sufficient, and even a single-step procedure has been utilized. In the latter case, analysis times are longer because of the presence of late-eluting ECD-active interferences. The two-step clean-up procedure generally used involves a liquid-liquid extraction with dichloromethane, and solid-liquid sorption using a Sep Pak C18 cartridge. PCP recovery over the 0.2-10 ppm range is 75-100%. Several wood samples containing 1-50 ppm of PCP have successfully been analyzed, and the good potential of NPLC-ECD for trace-level determination of PCP has been demonstrated.

  12. Mannheimiose pulmonar experimental em bezerros: swab nasal e nasofaringeano como auxílio diagnóstico Experimental pneumonic mannheimiosis in calves: nasal and nasopharingeal swabs for diagnostic

    Directory of Open Access Journals (Sweden)

    Adriana de Souza Coutinho

    2009-01-01

    Full Text Available Um modelo experimental de mannheimiosepneumônica bovina (MPB foi utilizado com o objetivo de avaliar as espécies bacterianas das cavidades nasais e nasofaringeanas em diferentes momentos do curso da doença, bem como verificar a eficiência diagnóstica do exame microbiológico dos swabs nasais (SN e nasofaringeanos (SNF. Um total de 28 bezerros foi distribuído aleatoriamente em quatro grupos experimentais (G1 a G4. SN e SNF foram colhidos sete dias antes e 12 (G1, 24 (G2, 48 (G3 e 72 (G4 horas após a inoculação intrabronquial de Mannheimia haemolytica. Após a indução da MPB, a bactéria M. haemolytica biotipo A foi predominante nos SN e SNF, sendo isolada em todos os momentos avaliados, com exceção de um SN colhido 24 horas após a indução da infecção. Não houve diferença significativa nas taxas de isolamento de Pasteurella multocida nos SN ou SNF, colhidos antes e após a indução da MPB. Contudo, esta bactéria passou a ser isolada mais freqüentemente após a indução da MPB, principalmente no SNF. Portanto, pode-se concluir que o exame microbiológico de SN e SNF é um teste auxiliar no diagnóstico da MPB.An experimental model of bovine pneumonic mannheimiosis (BPM was used to evaluate the nasal and nasopharynx bacterial species of calves during the course of the disease and for checking the diagnostic efficiency of nasal swab (NS and nasopharingeal swab (NPS microbiological exams. A total of 28 calves were randomized into four experimental groups (G1-G4. NS and NPS were obtained 7 days before and 12 (G1, 24 (G2, 48 (G3 e 72 (G4 hours after intrabronchial inoculation of Mannheimia haemolytica. After the induction of BPM, M. haemolytica biotype A was the predominant isolated bacterium in NS and NPS in all evaluated sampling times, except for one NS (harvested 24 hours. There were no significant statistical differences for the rates of Pasteurella multocida isolation in NS and NPS, harvested before and after the induction

  13. Comparison of illumigene Group A Streptococcus Assay with Culture of Throat Swabs from Children with Sore Throats in the New Zealand School-Based Rheumatic Fever Prevention Program.

    Science.gov (United States)

    Upton, Arlo; Bissessor, Liselle; Farrell, Elizabeth; Shulman, Stanford T; Zheng, Xiaotian; Lennon, Diana

    2016-01-01

    Group A streptococcal (GAS) pharyngitis is a particularly important condition in areas of New Zealand where the incidence of acute rheumatic fever remains unacceptably high. Prompt diagnosis and treatment of GAS pharyngitis are cornerstones of the Rheumatic Fever Prevention Programme, but these are hindered by the turnaround time of culture. Tests with excellent performance and rapid turnaround times are needed. For this study, throat swabs (Copan ESwabs) were collected from schoolchildren self-identifying with a sore throat. Samples were tested by routine culture and the illumigene GAS assay using loop-mediated isothermal amplification. Discrepant results were resolved by retesting of the same specimen by an alternative molecular assay. Seven hundred fifty-seven throat swab specimens were tested by both methods. The performance characteristics of the illumigene assay using culture on blood agar as the "gold standard" and following discrepancy analysis were as follows: sensitivity, 82% and 87%, respectively; specificity, 93% and 98%, respectively; positive predictive value, 61% and 88%, respectively; and negative predictive value, 97% and 97%, respectively. In our unique setting of a school-based throat swabbing program, the illumigene assay did not perform quite as well as described in previous reports. Despite this, its improved sensitivity and rapid turnaround time compared with those of culture are appealing. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  14. Genetic Characterization of Fungi Isolated from the Environmental Swabs collected from a Compounding Center Known to Cause Multistate Meningitis Outbreak in United States Using ITS Sequencing

    Directory of Open Access Journals (Sweden)

    Irshad M. Sulaiman

    2014-08-01

    Full Text Available A multistate fungal meningitis outbreak started in September of 2012 which spread in 20 states of the United States. The outbreak has been fatal so far, and has affected 751 individuals with 64 deaths among those who received contaminated spinal injections manufactured by a Compounding Center located in Massachusetts. In a preliminary study, Food and Drug Administration (FDA investigated the outbreak in collaboration with Centers for Disease Control and Prevention (CDC, state and local health departments, and identified four fungal and several bacterial contaminations in the recalled unopened injection vials. This follow-up study was carried out to assess DNA sequencing of the ITS1 region of rRNA gene for rapid identification of fungal pathogens during public health outbreak investigations. A total of 26 environmental swabs were collected from several locations at the manufacturing premises of the Compounding Center known to have caused the outbreak. The swab samples were initially examined by conventional microbiologic protocols and a wide range of fungal species were recovered. Species-identification of these microorganisms was accomplished by nucleotide sequencing of ITS1 region of rRNA gene. Analysis of data confirmed 14 additional fungal species in the swabs analyzed.

  15. HIV-1 and herpes simplex virus type-2 genital shedding among co-infected women using self-collected swabs in Chiang Rai, Thailand.

    Science.gov (United States)

    Forhan, S E; Dunne, E F; Sternberg, M R; Whitehead, S J; Leelawiwat, W; Thepamnuay, S; Chen, C; Evans-Strickfaden, Tt; McNicholl, J M; Markowitz, L E

    2012-08-01

    We analysed 528 genital self-collected swabs (SCS) from 67 HIV-1 and herpes simplex virus type-2 (HSV-2) co-infected women collected during the placebo month of a randomized crossover clinical trial of suppressive acyclovir in Chiang Rai, Thailand. In this first longitudinal study of HIV-1 and HSV-2 co-infected women using genital SCS specimens, we found frequent mucosal HIV-1 shedding. Overall, 372 (70%) swabs had detectable HIV-1 RNA with median HIV-1 viral load of 2.61 log(10) copies/swab. We found no statistically significant association between detectable HIV-1 RNA and HSV-2 DNA in the same SCS specimen (adjusted odds ratio [aOR] 1.40; 95% confidence intervals [CI], 0.78-2.60, P = 0.25). Only baseline HIV-1 plasma viral load was independently associated with genital HIV-1 RNA shedding (aOR, 7.6; 95% CI, 3.3-17.2, P genital sampling, and inclusion of genital sites other than the cervix.

  16. A Study of Isolates from Female Genital Swab Specimens in a ...

    African Journals Online (AJOL)

    To detect some common microbial agents of vaginal discharge in order to improve the current syndromic management of abnormal vaginal discharge. A prospective study of female genital swabs collected from Obstetrics and Gynecology units of Aminu Kano Teaching Hospital, Kano Nigeria and analyzed for microscopy, ...

  17. Evaluation of the reliability of Levine method of wound swab for ...

    African Journals Online (AJOL)

    The aim of this paper is to evaluate the reliability of Levine swab in accurate identification of microorganisms present in a wound and identify the necessity for further studies in this regard. Methods: A semi structured questionnaire was administered and physical examination was performed on patients with chronic wounds ...

  18. Acceptability of Sexually Transmitted Infection Testing Using Self-Collected Vaginal Swabs among College Women

    Science.gov (United States)

    Fielder, Robyn L.; Carey, Kate B.; Carey, Michael P.

    2013-01-01

    Objective: To assess the acceptability of sexually transmitted infection (STI) testing using self-collected vaginal swabs (SCVS) among college women. Participants: First-year female students ("N" = 483). Methods: Participants were offered free testing for 3 STIs using SCVS in April 2010 and later completed a survey regarding their…

  19. Mechanism of electron multiplication due to charging for a SiO2sample with a buried microstructure in SEM: A simulation analysis.

    Science.gov (United States)

    Wang, Fang; Feng, Guobao; Zhang, Xiusheng; Cao, Meng

    2016-11-01

    This study investigates the mechanism of electron redistribution and multiplication for a SiO 2 sample with a buried structure in scanning electron microscopy by numerical simulation. The simulation involved electron scattering and internal charge transport in the sample, the tracking of emitted secondary electrons (SEs), and the generation of tertiary electrons (TEs) produced by returned SEs due to charging of the sample. The results show that a buried grounded structure causes a non-uniform distribution of surface potential, and an electric field above the surface. As a result, although the number of escaped SEs above the margin of the buried structure decreases, the number of generated TEs increases more, leading to a final current of electrons that include escaped SEs and increased TEs. This multiplication of SEs might make a crucial contribution to the abnormal negative-charging contrast in SEM. During the electron beam irradiation, the variation in the number of total escaped electrons presents an obvious increase after an initial slight decrease, which corresponded to the transient characteristics of gray levels in SEM images from dark to abnormally bright. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Detection of newly produced T and B lymphocytes by digital PCR in blood stored dry on nylon flocked swabs.

    Science.gov (United States)

    Tessitore, Marion Vaglio; Sottini, Alessandra; Roccaro, Aldo M; Ghidini, Claudia; Bernardi, Simona; Martellosio, Giovanni; Serana, Federico; Imberti, Luisa

    2017-04-05

    A normal number of T-cell receptor excision circles (TRECs) and K-deleting recombination excision circles (KRECs) is considered a biomarker for adequate new T- and B-cell production. In newborns, detection of TRECs and KRECs by real time PCR from dried blood spotted on filter paper is used for the screening of severe immunodeficiency. In adults, elderly and during diseases, where the number of TRECs is lower than in newborns and children, a large amount of DNA and a sensitive method of amplification are necessary to identify newly produced lymphocytes. DNA was prepared from blood of 203 healthy adults (range: 18-91 years old) absorbed for 10 s on flocked swabs and let to dry, or from peripheral blood mononuclear cells. DNA was subjected to digital PCR and to well established conventional real time PCR-based method using TREC- and KREC-specific primers and probes. The number of TRECs and KRECs was expressed per mL of blood. Statistical analysis was performed by nested ANOVA, Pearson coefficient of determination, and by linear regression tests. The novel method for the storage of dried blood on nylon flocked swabs and the use of digital PCR allow quantification of TRECs and KRECs with high degree of sensitivity, specificity, accuracy, and precision. TRECs and KRECs were amplified by digital PCR in all tested blood samples, including those obtained from elderly individuals (>70 years old) and that were negative by real time PCR. Furthermore, values of TRECs and KRECs obtained by digital PCR were in the range of those acquired by real time PCR. Our findings demonstrate that DNA isolation from dried blood on flocked swabs followed by digital PCR-based analysis represents a useful tool for studying new lymphocyte production in adults and elderly individuals. This suggests the potential use of the methodology when monitoring of clinical variables is limited by the number of molecules that can be amplified and detected, such as in patients with immunodeficiency or under

  1. [Antibiotic sensitivity of beta-hemolytic streptococci isolated from throat swabs and purulent material].

    Science.gov (United States)

    Jachna-Sawicka, Katarzyna; Pietrzak, Anna; Bogiel, Tomasz; Gospodarek, Eugenia

    2010-01-01

    The aim of this study was to evaluate the prevalence and susceptibility of beta-hemolytic streptococci isolated from throat swabs (142--29.9%) and purulent material (333--70.1%) taken from patients treated at University Hospital dr. A. Jurasz in Bydgoszcz Collegium Medicum. L. Rydygier in Bydgoszcz, Nicolaus Copernicus University in Torun in 2005-2009. Of the 475 tested strains, 156 (32.8%) were identified as S. pyogenes. This species accounted for 38.8% of strains isolated from purulent material and 19.0% of swabs from the throat. Among the strains isolated from throat swabs of 62 (43.7%) were identified as Streptococcus group C. Only 5.1% strains were identified as Streptococcus group F. All strains of beta-hemolytic streptococci were susceptible to ampicillin or penicillin, fluoroquinolones, vancomycin and linezolid. Erythromycin-susceptible strains was 83.8%, and 89.1% for clindamycin. A total of 51.3% of erythromycin resistance strains had the cMLS(B) phenotype (63.3% for strains from throat swabs and 46.3% of the purulent materials). Sensitivity to tetracycline was characterized by 51.2% of strains of beta-hemolytic streptococci. The percentage of strains susceptible to this antibiotic among isolates from throat swabs was 63.1%, and purulent material--48.0%. The lowest percentage of strains susceptible to tetracycline (14.1%) were found among S. agalactiae and Streptococcus group G (33.6%) strains. During the study time, saw an increase in the percentage of strains susceptible to tetracycline and erythromycin.

  2. Assessing the ability of swab data to determine the true burden of infection for the amphibian pathogen Batrachochytrium dendrobatidis.

    Science.gov (United States)

    Clare, Frances; Daniel, Olivia; Garner, Trent; Fisher, Matthew

    2016-06-01

    Batrachochytrium dendrobatidis (Bd) is a pathogenic fungus which causes the disease chytridiomycosis in amphibians by infecting the animals' epidermis. The most commonly applied method for the detection of Bd is the use of a sterile swab, rubbed over the keratinized areas of an amphibian and then processed to yield DNA for detection by qPCR. This method has been used to infer a threshold of lethal infection in some species; however, how reliable and reproducible the swabbing method is at detecting the true burden of infection suffered by individuals is not known. European midwife toads, Alytes obstetricans, are susceptible to chytridiomycosis and are highly parasitised by Bd across Europe. By quantifying Bd-load throughout the entire skin and comparing this to swab results taken from the same individual, we determined whether epidermal swabs provide a quantifiable and accurate indication of the true fungal burden suffered. Further, we examined whether we could infer a threshold for lethal infection based on comparison of swab data taken from infected A. obstetricans exhibiting different clinical states. From swab data, we detected significantly higher fungal burdens from moribund metamorphs compared to visually healthy individuals; however, the ability of these swab data to provide an accurate indication of the true fungal burden was not reliable. These data suggest that fungal load dynamics play an important role in disease-induced mortality in A. obstetricans at these sites, but that using swab data to infer an exact threshold for Bd-associated mortality might be inappropriate and misleading.

  3. Sample Preparation of Nano-sized Inorganic Materials for Scanning Electron Microscopy or Transmission Electron Microscopy: Scientific Operating Procedure SOP-P-2

    Science.gov (United States)

    2015-07-01

    sample. Nano- sized particles have a tendency to agglomerate during sample preparation. ERDC/GSL SR-15-1 4 3 Scope This SOP is used to determine the...conductive vs. nonconductive samples. The preparation of nanomaterial samples for imaging can be challenging as these materials tend to agglomerate or...aggregated or agglomerated samples. Another way is to extract the material from the liquid. In selected cases, imaging of the nanoparticles is aided

  4. Comparison of eight methods for the extraction of Bacillus atrophaeus spore DNA from eleven common interferents and a common swab.

    Directory of Open Access Journals (Sweden)

    Helen L Rose

    Full Text Available Eight DNA extraction products or methods (Applied Biosystems PrepFiler Forensic DNA Extraction Kit; Bio-Rad Instagene Only, Bio-Rad Instagene & Spin Column Purification; EpiCentre MasterPure DNA & RNA Kit; FujiFilm QuickGene Mini80; Idaho Technologies 1-2-3 Q-Flow Kit; MoBio UltraClean Microbial DNA Isolation Kit; Sigma Extract-N-Amp Plant and Seed Kit were adapted to facilitate extraction of DNA under BSL3 containment conditions. DNA was extracted from 12 common interferents or sample types, spiked with spores of Bacillus atropheaus. Resulting extracts were tested by real-time PCR. No one method was the best, in terms of DNA extraction, across all sample types. Statistical analysis indicated that the PrepFiler method was the best method from six dry powders (baking, biological washing, milk, plain flour, filler and talcum and one solid (Underarm deodorant, the UltraClean method was the best from four liquids (aftershave, cola, nutrient broth, vinegar, and the MasterPure method was the best from the swab sample type. The best overall method, in terms of DNA extraction, across all sample types evaluated was the UltraClean method.

  5. Surge and swab pressures in wells with cross-section changes; Pressoes de surge and swab em pocos com variacao de secao

    Energy Technology Data Exchange (ETDEWEB)

    Fedevjcyk, Joao Victor; Junqueira, Silvio Luiz de Mello; Negrao, Cezar Otaviano Ribeiro [Universidade Tecnologica Federal do Parana (UTFPR). Laboratorio de Ciencias Termicas (Lacit) (Brazil)], e-mails: silvio@utfpr.edu.br, negrao@utfpr.edu.br

    2009-12-15

    Well drilling is performed by rotating and applying a weighted drill bit to the geological formation. Well diameter variations and the use of drill pipe accessories might cause changes to the annular cross section space between the drill pipe and the borehole. It should be noted cross section changes influence pressure losses within the well. This study proposes a mathematical/ numerical model to simulate the surge and swab problem in wells with variable cross section areas. The fluid flow yielded by the drill pipe motion is considered to be one-dimensional, isothermal, compressible and transient. The proposed model features the mass and momentum conservation equations, along with a state equation and a constitutive equation for Bingham or Power Law fluids. The governing equations were discretized by the Finite Volume Method. The well is assumed to be impermeable and the drill pipe end to be closed. The results were compared to measured data obtained at the Taquipe experimental well with good agreement. Predictions can now be made as to how changes in cross section areas may significantly affect the transient surge and swab pressures. (author)

  6. Standards for the management of swabs, needles and instruments in the operating theatre

    Directory of Open Access Journals (Sweden)

    A. Astrop

    1996-05-01

    Full Text Available The management of swabs, needles and instruments in the operating theatre is a high-risk and problem-prone area for the operating theatre nurse. The purpose of this research is to formulate specific standards on the management of swabs, needles and instruments in the operating theatre to ensure the safety of the patient. An exploratory and descriptive research design was used and executed in 3 hospitals of a private hospital group in Gauteng. A structured two phase process was followed, ie the development phase and the validation phase. This last phase was done by means of deliberate debate. It is recommended that these standards be implemented, tested and validated on a national basis and a monitoring and evaluation system should be developed to ensure nursing compliance with these standards.

  7. Absence of Neisseria meningitidis from throat swabs of Kuwaiti pilgrims after returning from the Hajj.

    Science.gov (United States)

    Husain, Entesar H; Dashti, Ali A; Electricwala, Qudsiya Y; Abdulsamad, Abdulsamad M; Al-Sayegh, Safeya

    2010-01-01

    To investigate whether or not Neisseria meningitidis was present in the throat of Kuwaiti pilgrims after returning from the Hajj. Throat swabs were taken from 177 participants 1 week after returning from the Hajj. The participants were asked about: associated medical conditions, meningococcal vaccination status and the intake of ciprofloxacin before leaving Mecca for Kuwait. There was no throat colonization with N. meningitidis on any of the throat swabs. Of the 177 pilgrims, 163 (92%) were vaccinated with meningococcal quadrivalent vaccine before leaving to Saudi Arabia. Ninety-seven of the pilgrims (83%) had received one dose of ciprofloxacin before leaving Mecca. The result showed that vaccination before leaving Kuwait and ciprofloxacin prophylaxis were effective in preventing throat colonization with Neisseria meningitidis. Copyright 2010 S. Karger AG, Basel.

  8. Cryo-electron Microscopy Structures of Expanded Poliovirus with VHHs Sample the Conformational Repertoire of the Expanded State.

    Science.gov (United States)

    Strauss, Mike; Schotte, Lise; Karunatilaka, Krishanthi S; Filman, David J; Hogle, James M

    2017-02-01

    By using cryo-electron microscopy, expanded 80S-like poliovirus virions (poliovirions) were visualized in complexes with four 80S-specific camelid VHHs (Nanobodies). In all four complexes, the VHHs bind to a site on the top surface of the capsid protein VP3, which is hidden in the native virus. Interestingly, although the four VHHs bind to the same site, the structures of the expanded virus differ in detail in each complex, suggesting that each of the Nanobodies has sampled a range of low-energy structures available to the expanded virion. By stabilizing unique structures of expanded virions, VHH binding permitted a more detailed view of the virus structure than was previously possible, leading to a better understanding of the expansion process that is a critical step in infection. It is now clear which polypeptide chains become disordered and which become rearranged. The higher resolution of these structures also revealed well-ordered conformations for the EF loop of VP2, the GH loop of VP3, and the N-terminal extensions of VP1 and VP2, which, in retrospect, were present in lower-resolution structures but not recognized. These structural observations help to explain preexisting mutational data and provide insights into several other stages of the poliovirus life cycle, including the mechanism of receptor-triggered virus expansion. When poliovirus infects a cell, it undergoes a change in its structure in order to pass RNA through its protein coat, but this altered state is short-lived and thus poorly understood. The structures of poliovirus bound to single-domain antibodies presented here capture the altered virus in what appear to be intermediate states. A careful analysis of these structures lets us better understand the molecular mechanism of infection and how these changes in the virus lead to productive-infection events. Copyright © 2017 American Society for Microbiology.

  9. Cryo-electron Microscopy Structures of Expanded Poliovirus with VHHs Sample the Conformational Repertoire of the Expanded State

    Science.gov (United States)

    Strauss, Mike; Schotte, Lise; Karunatilaka, Krishanthi S.; Filman, David J.

    2016-01-01

    ABSTRACT By using cryo-electron microscopy, expanded 80S-like poliovirus virions (poliovirions) were visualized in complexes with four 80S-specific camelid VHHs (Nanobodies). In all four complexes, the VHHs bind to a site on the top surface of the capsid protein VP3, which is hidden in the native virus. Interestingly, although the four VHHs bind to the same site, the structures of the expanded virus differ in detail in each complex, suggesting that each of the Nanobodies has sampled a range of low-energy structures available to the expanded virion. By stabilizing unique structures of expanded virions, VHH binding permitted a more detailed view of the virus structure than was previously possible, leading to a better understanding of the expansion process that is a critical step in infection. It is now clear which polypeptide chains become disordered and which become rearranged. The higher resolution of these structures also revealed well-ordered conformations for the EF loop of VP2, the GH loop of VP3, and the N-terminal extensions of VP1 and VP2, which, in retrospect, were present in lower-resolution structures but not recognized. These structural observations help to explain preexisting mutational data and provide insights into several other stages of the poliovirus life cycle, including the mechanism of receptor-triggered virus expansion. IMPORTANCE When poliovirus infects a cell, it undergoes a change in its structure in order to pass RNA through its protein coat, but this altered state is short-lived and thus poorly understood. The structures of poliovirus bound to single-domain antibodies presented here capture the altered virus in what appear to be intermediate states. A careful analysis of these structures lets us better understand the molecular mechanism of infection and how these changes in the virus lead to productive-infection events. PMID:27852863

  10. Comparison between Saliva and Nasopharyngeal Swab Specimens for Detection of Respiratory Viruses by Multiplex Reverse Transcription-PCR.

    Science.gov (United States)

    Kim, Young-Gon; Yun, Seung Gyu; Kim, Min Young; Park, Kwisung; Cho, Chi Hyun; Yoon, Soo Young; Nam, Myung Hyun; Lee, Chang Kyu; Cho, Yun-Jung; Lim, Chae Seung

    2017-01-01

    Nasopharyngeal swabs (NPSs) are being widely used as specimens for multiplex real-time reverse transcription (RT)-PCR for respiratory virus detection. However, it remains unclear whether NPS specimens are optimal for all viruses targeted by multiplex RT-PCR. In addition, the procedure to obtain NPS specimens causes coughing in most patients, which possibly increases the risk of nosocomial spread of viruses. In this study, paired NPS and saliva specimens were collected from 236 adult male patients with suspected acute respiratory illnesses. Specimens were tested for 16 respiratory viruses by multiplex real-time RT-PCR. Among the specimens collected from the 236 patients, at least 1 respiratory virus was detected in 183 NPS specimens (77.5%) and 180 saliva specimens (76.3%). The rates of detection of respiratory viruses were comparable for NPS and saliva specimens (P = 0.766). Nine virus species and 349 viruses were isolated, 256 from NPS specimens and 273 from saliva specimens (P = 0.1574). Adenovirus was detected more frequently in saliva samples (P saliva samples was excluded by direct sequencing. In conclusion, neither of the sampling methods was consistently more sensitive than the other. We suggest that these cost-effective methods for detecting respiratory viruses in mixed NPS-saliva specimens might be valuable for future studies. Copyright © 2016 American Society for Microbiology.

  11. The effect of sample matrix on electron density, electron temperature and gas temperature in the argon inductively coupled plasma examined by Thomson and Rayleigh scattering

    Science.gov (United States)

    Hanselman, D. S.; Sesi, N. N.; Huang, M.; Hieftje, G. M.

    1994-05-01

    Spatially-resolved electron temperature ( Te), electron number density ( ne) and gas-kinetic temperature ( Tg) maps of the inductively coupled plasma (ICP) have been obtained for two central-gas flow rates, four heights above the load coil (ALC) and in the presence and absence of interferants with a wide range of first ionization potentials. The radial profiles demonstrate how the directly measured fundamental parameters neTe and Tg can be significantly enhanced and/or depressed with added interferent, depending upon plasma operating conditions and observation region. In general, the magnitude of ne, and Te change is found to be an inverse function of interferent ionization potential; furthermore, ne enhancements in the central channel might be the result of electron redistribution from high to low electron density regions rather than from ionization of the matrix. The large measured increases in ne cannot be attributed solely to matrix ionization, especially when measurement uncertainties and the probable over-estimation in calculated ne, enhancements are taken into account. Changes in ne and Te have been correlated with axial Ca atom and ion emission profiles. A brief review of the mechanisms most likely involved in interelement matrix interferences is given within the context of the present study. This article is an electronic publication in Spectrochimica Acta Electronica (SAE), the electronic section of Spectrochimica Acta Part B (SAB). The hardcopy text is accompanied by a disk for the Macintosh computer with data files stored in ASCII format. The main article discusses the scientific aspects of the subject and gives an interpretation of the results contained in the data files.

  12. Evaluation of a PCR test for detection of treponema pallidum in swabs and blood.

    Science.gov (United States)

    Grange, P A; Gressier, L; Dion, P L; Farhi, D; Benhaddou, N; Gerhardt, P; Morini, J P; Deleuze, J; Pantoja, C; Bianchi, A; Lassau, F; Avril, M F; Janier, M; Dupin, N

    2012-03-01

    Syphilis diagnosis is based on clinical observation, serological analysis, and dark-field microscopy (DFM) detection of Treponema pallidum subsp. pallidum, the etiological agent of syphilis, in skin ulcers. We performed a nested PCR (nPCR) assay specifically amplifying the tpp47 gene of T. pallidum from swab and blood specimens. We studied a cohort of 294 patients with suspected syphilis and 35 healthy volunteers. Eighty-seven of the 294 patients had primary syphilis, 103 had secondary syphilis, 40 had latent syphilis, and 64 were found not to have syphilis. The T. pallidum nPCR results for swab specimens were highly concordant with syphilis diagnosis, with a sensitivity of 82% and a specificity of 95%. Reasonable agreement was observed between the results obtained with the nPCR and DFM methods (kappa = 0.53). No agreement was found between the nPCR detection of T. pallidum in blood and the diagnosis of syphilis, with sensitivities of 29, 18, 14.7, and 24% and specificities of 96, 92, 93, and 97% for peripheral blood mononuclear cell (PBMC), plasma, serum, and whole-blood fractions, respectively. HIV status did not affect the frequency of T. pallidum detection in any of the specimens tested. Swab specimens from mucosal or skin lesions seemed to be more useful than blood for the efficient detection of the T. pallidum genome and, thus, for the diagnosis of syphilis.

  13. On the role of the gas environment, electron-dose-rate, and sample on the image resolution in transmission electron microscopy

    DEFF Research Database (Denmark)

    Ek, Martin; Jespersen, Sebastian Pirel Fredsgaard; Damsgaard, Christian Danvad

    2016-01-01

    The introduction of gaseous atmospheres in transmission electron microscopy offers the possibility of studying materials in situ under chemically relevant environments. The presence of a gas environment can degrade the resolution. Surprisingly, this phenomenon has been shown to depend on the elec...

  14. Simplicimonas-like DNA in vaginal swabs of cows and heifers cross-reacting in the real-time PCR for T. foetus.

    Science.gov (United States)

    Frey, Caroline F; Müller, Norbert; Stäuber, Norbert; Marreros, Nelson; Hofmann, Larissa; Hentrich, Brigitte; Hirsbrunner, Gaby

    2017-04-15

    Cows on an alpine pasture were presented with severe signs of vaginitis. To rule out infection with Tritrichomonas foetus, vaginal swabs were taken and real-time PCR based on detection via fluorescence resonance energy transfer (FRET) probes and targeting the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA (rDNA) was performed. PCR was positive in 25 of totally 34 assessed cows. However, the melting profiles of the probes targeting the diagnostic PCR products differed from the T. foetus positive control. Subsequent sequencing of the amplicons revealed 91% identity to Simplicimonas sp. sequences deposited in GenBank™. Furthermore, there was no clear association between positive PCR result and presence of vaginitis. To investigate the distribution of this Simplicimonas-like organism in cows, more herds grazing on the same alpine pastures as well as unrelated cows were tested. In total, 133 cows and 16 heifers were sampled, 53 cows and 6 heifers even twice. Vaginitis was evident in 43 cows and 4 heifers. All-over-positivity of PCR was 44%, including nine tests performed on heifers. Melting peak analysis indicated Simplicimonas-like organisms in all positive samples. Culture attempts in bovine InPouch ™ TF failed. No association between a positive PCR result and the presence of vaginitis was found. This is, to the best of our knowledge, the first report on Simplicimonas-like DNA in vaginal swabs of female cattle. Our data suggest that when testing vaginal swabs of cattle by means of T. foetus PCR, false positive reactions due to Simplicimonas-like organisms may occur. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Detection and genetic characterization of foot‐and‐mouth disease viruses in samples from clinically healthy animals in endemic settings

    DEFF Research Database (Denmark)

    Jamal, Syed Muhammad; Ferrari, G.; Hussain, M.

    2012-01-01

    A total of 1501 oral swab samples from Pakistan, Afghanistan and Tajikistan were collected from clinically healthy animals between July 2008 and August 2009 and assayed for the presence of foot‐and‐mouth disease virus (FMDV) RNA. The oral swab samples from two (of four) live animal markets in Pak...

  16. ANTIBIOTIC RESISTANCE IN ENTEROBACTERIACEAE STRAINS ISOLATED FROM CHICKEN AND MILK SAMPLES

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2015-02-01

    Full Text Available Antibiotic resistance and identification of strains in Enterobacteriaceae genera isolated from milk, milk products and rectal swabs of chicken was examined in this experiment. After samples collection cultivation and identification of bacterial strain was done. MALDI TOF MS Biotyper for identification of Enterobacteriaceae strains was used. For susceptibility testing disc diffusion methodology was used according by EUCAST. Results showed high level of ampicillin resistance in isolates from milk and milk samples. The highest streptomycin resistance was detected in isolates from rectal swabs of chicken. After identification, we determined that S. enterica ser. Typhimurium, which was isolated from rectal swabs of chicken showed the most multi-resistance from all identificated strains of Enterobacteriaceae. The most isolates bacterial strain was E. coli, which showed resistance against four antibiotics from rectal swabs of chicken. Also our results showed that the higher resistance level is in rectal swabs of chicken like in milk samples.

  17. Cryogenic coherent X-ray diffraction imaging of biological samples at SACLA: a correlative approach with cryo-electron and light microscopy.

    Science.gov (United States)

    Takayama, Yuki; Yonekura, Koji

    2016-03-01

    Coherent X-ray diffraction imaging at cryogenic temperature (cryo-CXDI) allows the analysis of internal structures of unstained, non-crystalline, whole biological samples in micrometre to sub-micrometre dimensions. Targets include cells and cell organelles. This approach involves preparing frozen-hydrated samples under controlled humidity, transferring the samples to a cryo-stage inside a vacuum chamber of a diffractometer, and then exposing the samples to coherent X-rays. Since 2012, cryo-coherent diffraction imaging (CDI) experiments have been carried out with the X-ray free-electron laser (XFEL) at the SPring-8 Ångstrom Compact free-electron LAser (SACLA) facility in Japan. Complementary use of cryo-electron microscopy and/or light microscopy is highly beneficial for both pre-checking samples and studying the integrity or nature of the sample. This article reports the authors' experience in cryo-XFEL-CDI of biological cells and organelles at SACLA, and describes an attempt towards reliable and higher-resolution reconstructions, including signal enhancement with strong scatterers and Patterson-search phasing.

  18. Screening for methicillin-resistant Staphylococcus aureus in clinical swabs using a high-throughput real-time PCR-based method

    DEFF Research Database (Denmark)

    Ornskov, D; Kolmos, B; Bendix Horn, P

    2008-01-01

    The presence of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals and the community is a serious problem. Accordingly, a comprehensive plan has been implemented in the County of Vejle, Denmark, to identify colonised and/or infected individuals and to control the spread of MRSA. Since...... 2005, all patients and healthcare personnel have been screened for MRSA colonisation, involving analysis of 300-400 samples daily. To deal with this number of samples, a PCR-based method customised for high-throughput analysis and a system for fast reporting of MRSA carrier status were developed. Swab...... samples were incubated overnight in a selective tryptone soya broth and were analysed by PCR the following day. Using this strategy, non-colonised individuals were identified within 24 h, while MRSA-positive samples were analysed further by traditional microbiological methods to determine the resistance...

  19. Blotting-free and lossless cryo-electron microscopy grid preparation from nanoliter-sized protein samples and single-cell extracts.

    Science.gov (United States)

    Arnold, Stefan A; Albiez, Stefan; Bieri, Andrej; Syntychaki, Anastasia; Adaixo, Ricardo; McLeod, Robert A; Goldie, Kenneth N; Stahlberg, Henning; Braun, Thomas

    2017-03-01

    We present a sample preparation method for cryo-electron microscopy (cryo-EM) that requires only 3-20nL of sample to prepare a cryo-EM grid, depending on the protocol used. The sample is applied and spread on the grid by a microcapillary. The procedure does not involve any blotting steps, and real-time monitoring allows the water film thickness to be assessed and decreased to an optimum value prior to vitrification. We demonstrate that the method is suitable for high-resolution cryo-EM and will enable alternative electron microscopy approaches, such as single-cell visual proteomics. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  20. The use of buccal swabs as a minimal-invasive method for detecting effects of pesticide exposure on enzymatic activity in common wall lizards.

    Science.gov (United States)

    Mingo, Valentin; Lötters, Stefan; Wagner, Norman

    2017-01-01

    Habitat loss and environmental pollution are among the main causes responsible for worldwide biodiversity loss. The resulting species and population declines affect all vertebrates including reptiles. Especially in industrialized countries, pollution by agrochemicals is of remarkable importance. Here, habitat loss has historically been associated with expansion of agriculture. Species persisting in such environments do not only need to cope with habitat loss, but more recently, also with chemical intensification, namely pesticide exposure. In this study, we examined effects of different fungicide and herbicide applications on the common wall lizard (Podarcis muralis) in grape-growing areas. We used three enzymatic biomarkers (GST, GR, AChE) and for the first time saliva from buccal swabs as a minimal-invasive sampling method for detection. Our results demonstrate absorption of substances by lizards and effects of pesticide exposure on enzymatic activities. Our findings are in accordance with those of previous laboratory studies, although samples were retrieved from natural habitats. We conclude that buccal swabs could become a useful tool for the detection of pesticide exposure in reptiles and have the potential to replace more invasive methods, such as organ extraction or cardiac puncture. This is an important finding, as reptiles are non-target organisms of pesticide applications, and there is a strong need to integrate them into pesticide risk assessments. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Screening femoral heads from living donors: a prospective study comparing swab-agar versus bone fragment-broth culture.

    Science.gov (United States)

    Saegeman, Veroniek; Verhaegen, Jan; Simon, Jean-Pierre

    2011-06-01

    Femoral heads are an important source of allograft bone used in reconstructive orthopaedic surgery. The sterility of donor material is of major importance for the recipient. Femoral heads intraoperatively retrieved during hip arthroplasty from medically screened living donors are routinely checked with a surface swab to exclude microbiological contamination. There is, however, evidence that swab cultures have limited sensitivity. We therefore prospectively compared two ways of screening femoral heads. Bacterial recovery from swabs in Amies transport medium taken intraoperatively, subsequently transported to the microbiology laboratory and inoculated on agar and in broth was compared with the recovery from a bone fragment also taken intraoperatively but immediately inoculated into Wilkins Chalgren broth. Forty femoral heads were tested with both methods. Bacteria were cultured neither from the femoral surface swabs nor from the femoral fragments. Consequently no distinct conclusions regarding the sensitivity of both techniques could be drawn. In addition the bacterial yield of two swabs in Amies transport medium streaked on a variety of culture media other than the conventional agar plates was also studied. Culturing of these swabs resulted in the detection of bacteria that are predominantly considered contaminants.

  2. Observational study of Streptococcus pyogenes isolated from vaginal swabs of adult women in a hospital and community laboratory.

    Science.gov (United States)

    Upton, Arlo; Taylor, Susan

    2013-12-01

    Streptococcus pyogenes or group A streptococcus (GAS) is a common cause of vulvo-vaginitis in pre-pubertal females but is uncommonly isolated from the vaginal swabs of adult females. We aimed to describe the clinical and laboratory findings of adult females with GAS isolated from vaginal swabs in a community and hospital laboratory. Over a 19 week period the two laboratories identified females ≥ 15 years of age with GAS isolated from vaginal swabs. At least 2 weeks after reporting, the referring doctor or midwife was telephoned by the authors for clinical information or the clinical notes were reviewed. Laboratory data were also collected. One hundred adult females with GAS isolated from vaginal swabs were identified from approximately 4500-5000 community laboratory, and 20 from approximately 2000 hospital laboratory swabs. Community patients were more likely to have presented with vaginal symptoms such as discharge, while hospital patients were more likely to have ascending infection related to pregnancy/recent delivery. Of the community patients, 15% were asymptomatic compared with 5% of the hospital patients. Review of Gram stain and culture quantification was not found to be particularly useful for discriminating between clinical infection and asymptomatic colonisation. Isolation of GAS from the vaginal swabs of adult females is uncommon. In the community setting it may represent infection with vulvo-vaginitis or asymptomatic colonisation. In the hospital setting, its isolation is frequently associated with pregnancy-related infectious complications.

  3. Development for PRESPEC: New front end electronic for multi sampling ionization chambers of the GSI-fragment separator

    Energy Technology Data Exchange (ETDEWEB)

    Pietri, Stephane; Gerl, Juergen; Kurz, Nik; Nociforo, Chiara; Schaffner, Henning; Simon, Haik [GSI, Planckstrasse 1, 64291 Darmstadt (Germany)

    2009-07-01

    To study detailed structure effects during in-beam gamma spectroscopy experiments at the GSI fragment separator high particles rates are needed at the final focal plane. The use of new position detector (TPC) having higher rate capability than the previous Multi-Wire Proportional Counter put the ionization chambers (MUSICs) used for the {delta}-E measurement as the limiting factor. The current electronics used for those detector does not allow to sustain rates of up to 100 kHz that the coming experimental program will request. Indeed at those rates the analog electronic is not able to disentangle pile-up events and thus give a wrong Z identification for the incoming nucleus. The ongoing work on new digital electronic and on signal characterization to allow higher rates capability of those detectors is presented.

  4. Scaling down the two-dimensional electron gas spin resonance (ESR) phenomena in GaAs/AlGaAs heterostructures to sub-micron samples

    Science.gov (United States)

    Bandaru, Prabhakar; Yablonovitch, Eli; Jiang, Hong-Wen

    2002-03-01

    Electron Spin Resonance (ESR) has been proposed as a technique for achieving single electron and subsequently single spin control, important for the emerging fields of spintronics and quantum computing. In this paper, we report on ESR in the quantum Hall regime, of sub-micron structures containing a few hundred electrons. These phenomena are contrasted with ESR phenomena in structures containing 10^7 - 10^9 electrons, which have been performed so far (Reference 1). There are several novel features observed in the ESR of small structures, such as a very large decrease of resistance and shift in the quantum Hall minima to lower magnetic fields after the resonance. These imply a reduction in the number of electrons and could result from the greater influence of the surface and impurity potential fluctuations intrinsic to a small sample. The ESR peak intensity is hypothesized to result from the transfer of electrons from the localized states to the extended states. References: 1.H.W.Jiang and E. Yablonovitch, Phys. Rev.B., 64, R041307, (2001) 2.M.Dobers, K.v. Klitzing and G. Weimann,Phys. Rev. B, 38, 5453, (1988).

  5. Hidden in plain sight: bias towards sick patients when sampling patients with sufficient electronic health record data for research.

    Science.gov (United States)

    Rusanov, Alexander; Weiskopf, Nicole G; Wang, Shuang; Weng, Chunhua

    2014-06-11

    To demonstrate that subject selection based on sufficient laboratory results and medication orders in electronic health records can be biased towards sick patients. Using electronic health record data from 10,000 patients who received anesthetic services at a major metropolitan tertiary care academic medical center, an affiliated hospital for women and children, and an affiliated urban primary care hospital, the correlation between patient health status and counts of days with laboratory results or medication orders, as indicated by the American Society of Anesthesiologists Physical Status Classification (ASA Class), was assessed with a Negative Binomial Regression model. Higher ASA Class was associated with more points of data: compared to ASA Class 1 patients, ASA Class 4 patients had 5.05 times the number of days with laboratory results and 6.85 times the number of days with medication orders, controlling for age, sex, emergency status, admission type, primary diagnosis, and procedure. Imposing data sufficiency requirements for subject selection allows researchers to minimize missing data when reusing electronic health records for research, but introduces a bias towards the selection of sicker patients. We demonstrated the relationship between patient health and quantity of data, which may result in a systematic bias towards the selection of sicker patients for research studies and limit the external validity of research conducted using electronic health record data. Additionally, we discovered other variables (i.e., admission status, age, emergency classification, procedure, and diagnosis) that independently affect data sufficiency.

  6. Rapid detection of Van genes in rectal swabs by real time PCR in Southern Brazil

    Directory of Open Access Journals (Sweden)

    Vlademir Cantarelli

    2011-10-01

    Full Text Available INTRODUCTION: Laboratory-based surveillance is an important component in the control of vancomycin resistant enterococci (VRE. METHODS: The study aimed to evaluate real-time polymerase chain reaction (RT-PCR (genes vanA-vanB for VRE detection on 115 swabs from patients included in a surveillance program. RESULTS: Sensitivity of RT-PCR was similar to primary culture (75% and 79.5%, respectively when compared to broth enriched culture, whereas specificity was 83.1%. CONCLUSIONS: RT-PCR provides same day results, however it showed low sensitivity for VRE detection.

  7. Comparison of serum, ear notches, and nasal and saliva swabs for Bovine viral diarrhea virus antigen detection in colostrum-fed persistently infected (PI) calves and non-PI calves.

    Science.gov (United States)

    Lanyon, Sasha R; Sims, Sarah K; Cockcroft, Peter D; Reichel, Michael P

    2014-11-01

    The diagnosis of neonatal and young calves persistently infected (PI) with Bovine viral diarrhea virus (BVDV) by antigen-capture enzyme-linked immunosorbent assay (ACE) may be complicated by interference from colostrum-derived specific antibodies. Ten calves, with 3 calves identified as PI and 7 as non-PI were used in the current study. All non-PI calves were shown to be seropositive for BVDV-specific antibodies by antibody enzyme-linked immunosorbent assay (Ab-ELISA) on serum. Serum samples, ear notch samples, and nasal and saliva swabs were collected from each calf from birth until 12 weeks of age and tested by ELISA for BVDV-specific antigen and antibodies. Following colostrum ingestion, Ab-ELISA sample-to-positive (S/P) ratios rose by a mean of 0.95 (95% confidence interval [CI] = 0.64-1.25) and 1.72 (95% CI = 1.55-1.89) in seropositive, non-PI calves and in PI calves, respectively. The mean S/P ratios then declined to approximately 1.1 in non-PI calves and 0.5 in PI calves at between 60 and 80 days of age. In PI calves, testing for antigen in serum and nasal and saliva swabs was subject to interference by colostrum-derived antibodies in calves up to 3 weeks of age. Nasal swabs were less affected than serum and saliva swabs. Ear notches maintained positive ACE corrected optical densities at all sample times, despite a drop in the signal following the ingestion of colostrum. © 2014 The Author(s).

  8. Evaluation of Simplexa™ Group A Strep Direct Kit compared to Hologic® Group A streptococcal direct assay for detection of Group A Streptococcus (GAS) in throat swabs.

    Science.gov (United States)

    Church, Deirdre L; Lloyd, Tracie; Larios, Oscar; Gregson, Daniel B

    2018-01-05

    Diagnosis of bacterial pharyngitis is confirmed by detection of Group A Streptococcus (GAS) in patient throat samples. Testing of throat samples has historically relied on culture but new molecular methods allow much faster test turnaround time (i.e., same day vs. 48-72h for culture). Our laboratory uses the Hologic® GAS Direct (GASD) assay for screening more than 125,000 throat samples per annum. Simplexa™ GAS Direct is a new real-time PCR (qPCR) assay that does not require initial DNA extraction. Performance of Simplexa qPCR was compared to GASD. 289 throat swabs were collected from patients attending ambulatory clinics in Calgary. A total of 60 (20.8%) of the samples were initially GAS positive by either method; 54 by both methods, 4 by Simplex qPCR alone, and 2 by GASD alone. An in-house PCR using a unique GAS primer set was used to resolve the 6 discrepant results. Overall, GASD compared to Simplexa qPCR had sensitivity, specificity, positive predictive value and negative predictive value of 93.1% vs 100%, 100% vs. 100%, 100% vs. 100% and 98.31% vs. 100% respectively. Implementation of Simplexa qPCR in our laboratory setting would cost more but allow the high sample volume to be reported in half the time and save 0.62 MLT FTE. In comparison to culture, the implementation of Simplexa qPCR would save 2.79 MLA FTE plus 0.94 MLT FTE. Simplexa qPCR has improved performance and diagnostic efficiency in a high-volume laboratory compared to GASD for GAS detection in throat swabs. Copyright © 2018 American Society for Microbiology.

  9. Polymerase chain reaction for detection of Chlamydia trachomatis in conjunctival swabs

    Science.gov (United States)

    Elnifro, E.; Storey, C.; Morris, D.; Tullo, A.

    1997-01-01

    AIMS/BACKGROUND—Ocular Chlamydia trachomatis infection in the west occurs as ophthalmia neonatorum, acquired from the mother, or adult paratrachoma which is also associated with current genital tract infection. Accurate rapid laboratory diagnosis facilitates management, but the relative merits of antigen detection or DNA amplification tests are unresolved.
METHODS—A polymerase chain reaction (PCR) test was developed which amplified part of the plasmid shared by all the serovars of C trachomatis. Conjunctival swabs were tested using an in house immune dot-blot test (IDBT) for chlamydial lipopolysaccharide antigen, a commercial direct fluorescent antibody (DFA) test for chlamydial elementary bodies, and the PCR (DNA extracted using guanidinium lysis buffer).
RESULTS—The PCR achieved a detection limit of 100 plasmid copies (10 elementary bodies). In a combined retrospective and prospective clinical evaluation, the PCR and IDBT gave identical results with 21 positive and 57 negative eye swabs. However, interpretation of the DFA test required meticulous examination of the stained smear, sometimes by two microscopists.
CONCLUSIONS—The PCR is likely to play an increasing role in the diagnosis of ocular C trachomatis infection because of its excellent sensitivity and specificity.

 PMID:9274416

  10. Rapid analysis of pyrethroid insecticides in aquaculture seawater samples via membrane-assisted solvent extraction coupled with gas chromatography-electron capture detection

    OpenAIRE

    Shi, X.Z.; Song, S.Q.; Sun, A.L.; Liu, J.H.; Li, D.X.; Chen, J.

    2012-01-01

    A simple, efficient, and environmentally friendly membrane-assisted solvent extraction (MASE) method for the extraction and preconcentration of six pyrethroid insecticides from aquaculture seawater samples followed by gas chromatography-electron capture detection (GC-ECD) was successfully proposed. The operating conditions for MASE, such as the extraction solvent, solvent volume, NaCl concentration, stirring rate, extraction time, and temperature, were optimized. Compared to conventional Flor...

  11. Electrostatic sampling of trace DNA from clothing.

    Science.gov (United States)

    Zieger, Martin; Defaux, Priscille Merciani; Utz, Silvia

    2016-05-01

    During acts of physical aggression, offenders frequently come into contact with clothes of the victim, thereby leaving traces of DNA-bearing biological material on the garments. Since tape-lifting and swabbing, the currently established methods for non-destructive trace DNA sampling from clothing, both have their shortcomings in collection efficiency and handling, we thought about a new collection method for these challenging samples. Testing two readily available electrostatic devices for their potential to sample biological material from garments made of different fabrics, we found one of them, the electrostatic dust print lifter (DPL), to perform comparable to well-established sampling with wet cotton swabs. In simulated aggression scenarios, we had the same success rate for the establishment of single aggressor profiles, suitable for database submission, with both the DPL and wet swabbing. However, we lost a substantial amount of information with electrostatic sampling, since almost no mixed aggressor-victim profiles suitable for database entry could be established, compared to conventional swabbing. This study serves as a proof of principle for electrostatic DNA sampling from items of clothing. The technique still requires optimization before it might be used in real casework. But we are confident that in the future it could be an efficient and convenient contribution to the toolbox of forensic practitioners.

  12. Airborne inorganic fibre level monitoring by transmission electron microscope (TEM): comparison of direct and indirect sample transfer methods.

    Science.gov (United States)

    Sahle, W; Laszlo, I

    1996-02-01

    The direct and indirect sample transfer techniques for measuring airborne inorganic fibres concentrations were studied by TEM for airborne rockwool fibres, chrysotile, amosite and tungsten oxide whiskers. The number and mass concentrations of fibres with aspect ratios > or = 5, prepared by these two techniques for fibres longer than 0.5 mu m and 5 mu m, and the aspect ratios were evaluated and compared. It was found that the indirect sample transfer technique affects the fibre size distribution of different materials differently, and that the mass concentrations did not relate to fibre number concentrations of ambient air, irrespective of the sample transfer method used or the materials studied. It is concluded that the direct sample transfer method incorporating the etching stage should be preferred over the indirect method. Moreover, a need for an extra stage in-between the etching and carbon coating stages for the direct sample transfer method is suggested; this would be a stage where soluble materials could be removed.

  13. Development of a quantitative real time PCR assay to detect and enumerate Escherichia coli O157 and O26 serogroups in bovine recto-anal swabs.

    Science.gov (United States)

    Lawal, Dolapo; Burgess, Catherine; McCabe, Evonne; Whyte, Paul; Duffy, Geraldine

    2015-07-01

    Escherichia coli O157 and O26 shedding patterns in cattle are known to vary widely. To address gaps in the understanding of the underlying factors which impact on shedding dynamics, sensitive and rapid quantitative methods which can be applied in surveillance studies on cattle are required. Current approaches for enumeration of verocytotoxigenic E. coli (VTEC) in cattle faeces are based on direct plating onto selective agars, most probable number (MPN) or real time PCR applied directly to faecal samples, all of which have limitations in terms of the labour involved or their sensitivity. The objective of this study was to develop a sensitive real time quantitative PCR assay, to quantify O157 and O26 in bovine recto-anal junction (RAJ) swabs. The approach was to target serogroup specific genes rfbE and wzx, and to couple a short enrichment, with the use of a standard calibration curve relating real time PCR cycle threshold (Ct) values against the initial concentration of the pathogen in the sample. Following initial experiments in broth culture, a 5h enrichment in modified tryptone soya broth with novobiocin (20 mg/l) (mTSBn) was found to be optimal, and a linear correlation between inocula (Log10 1 to 6 CFU ml(-1)) and the PCR Ct values for both E. coli O157 (R(2)=0.99, rsd=0.58) and E. coli O26 (R(2)=0.99, rsd=0.44) was confirmed. The developed method was then applied to bovine RAJ swab samples (n=153), which were inoculated with E. coli O157 or O26 (Log10 1 to 7 CFU swab(-1)). Calibration curves yielded correlations for E. coli O157 of R(2)=0.86, rsd=0.72 and for O26 (R(2)=0.88, rsd=0.69). In conclusion, a sensitive method for detection and enumeration of two significant VTEC serogroups in bovine RAJ samples has been developed and validated, and will support studies on the bovine shedding dynamics of these pathogens in cattle. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Comparison of sputum and nasopharyngeal swab specimens for molecular diagnosis of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila.

    Science.gov (United States)

    Cho, Min-Chul; Kim, Hyewon; An, Dongheui; Lee, Miyoung; Noh, Shin-Ae; Kim, Mi-Na; Chong, Young Pil; Woo, Jun Hee

    2012-03-01

    Differentiation of atypical pathogens is important for community-acquired pneumonia (CAP). In this study, we compared sputum and nasopharyngeal swabs (NPS) for use in detection of Mycoplasma pneumoniae (MP), Chlamydophila pneumoniae (CP), and Legionella pneumophila (LP), using Seeplex PneumoBacter ACE Detection Assay (PneumoBacter; Seegene). Sputum and NPS specimens were collected from patients in 15 hospitals. DNA was extracted from sputum using QIAamp DNA Stool Mini Kit (Qiagen) and from NPS using easyMAG (bioMérieux). Both types of specimens were evaluated by multiplex PCR using PneumoBacter. To determine the diagnostic performance of this assay, sputum samples were also tested using BD ProbeTec ET Atypical Pneumonia Assay (APA; Becton Dickinson). Among 217 sputum and NPS, 20 (9.2%), 2 (0.9%), and 0 sputum were positive for MP, LP, and CP, respectively, whereas 8 (3.7%) NPS were positive for MP. The sputum APA test yielded 186, 206, and 204 interpretable results for MP, LP, and CP, respectively. Of these, 21 (11.3%) were positive for MP, 2 (1.0%) were positive for LP, and 0 samples were positive for CP. Compared to APA, the sensitivity and specificity of the sputum assay for MP were 95.2% and 100.0%, respectively, whereas for the NPS assay, these were 38.1% and 93.9%. Sputum testing was more sensitive than NPS testing (P=0.002). For LP and CP diagnosis, PneumoBacter and APA tests agreed 100%. Specimen type is crucial and sputum is preferred over NPS for simultaneous detection of MP, LP, and CP using multiplex PCR in CAP.

  15. Analysis of hexachlorocyclohexanes in aquatic samples by one-step microwave-assisted headspace controlled-temperature liquid-phase microextraction and gas chromatography with electron capture detection.

    Science.gov (United States)

    Tsai, Ming-Yuen; Kumar, Ponnusamy Vinoth; Li, Hong-Ping; Jen, Jen-Fon

    2010-03-19

    A microwave-assisted headspace controlled-temperature liquid-phase microextraction (HS-CT-LPME) technique was applied for the one-step sample extraction of hexachlorocyclohexanes (HCHs) from aqueous samples with complicate matrices, followed by gas chromatographic (GC) analysis with electron capture detector (ECD). Microwave heating was applied to accelerate the evaporation of HCHs into the headspace and an external-cooling system was used to control the temperature in the sampling zone for HS-LPME. Parameters affecting extraction efficiency, such as LPME solvent, sampling position and temperature, microwave power and irradiation time (the same as sampling time), sample pH, and salt addition were thoroughly investigated. From experimental results, the following conditions were selected for the extraction of HCHs from 10-mL water sample (pH 2.0) by using 1-octanol as the LPME solvent, with sampling done at 38 degrees C for 6 min under 167 W of microwave irradiation. The detections were linear in the concentration of 0.1-10 microg/L for alpha-HCH and gamma-HCH, and 1-100 microg/L for beta-HCH and delta-HCH. Detection limits were 0.05, 0.4, 0.03 and 0.1 microg/L for alpha-, beta-, gamma- and delta-HCH, respectively. Environmental water samples were analyzed with recovery between 86.4% and 102.4% for farm-field water, and between 92.2% and 98.6% for river water. The proposed method proved to serve as a simple, rapid, sensitive, inexpensive, and eco-friendly procedure for the determination of HCHs in aqueous samples. Copyright 2010 Elsevier B.V. All rights reserved.

  16. Use and Limitations of Electron Flood Gun Control of Surface Potential During XPS: Two Non-homogeneous Sample Types

    Energy Technology Data Exchange (ETDEWEB)

    Baer, Donald R.; Engelhard, Mark H.; Gaspar, Dan J.; Lea, Alan S.; Windisch, Charles F.

    2002-10-01

    The ability of charge compensation methods to control the surface potentials for two types of non-homogenous samples is examined. Results demonstrate that two newer types of charge compensation systems have improved performance in relation to some previous flood gun methods and reaffirm the concept that a primary objective of charge compensation is to find conditions for which the surface potential of the specimen is as uniform as possible. However, experiments involving both flood gun use and specimen grounding, demonstrate that peak broadening and shifting can occur when two (or more) potentials are present in the region of analysis. Finally, the ability of interface charge to shift specimen potentials and measured binding energies demonstrates fundamental limitations to the absolute accuracy of binding energy measurements, but also remind us that charging phenomena can be used to obtain important information about the sample.

  17. Human papillomavirus self-sampling for screening nonattenders: Opt-in pilot implementation with electronic communication platforms.

    Science.gov (United States)

    Lam, Janni Uyen Hoa; Rebolj, Matejka; Møller Ejegod, Ditte; Pedersen, Helle; Rygaard, Carsten; Lynge, Elsebeth; Thirstrup Thomsen, Louise; Krüger Kjaer, Susanne; Bonde, Jesper

    2017-05-15

    In organized cervical screening programs, typically 25% of the invited women do not attend. The Copenhagen Self-sampling Initiative (CSi) aimed to gain experiences on participation among screening nonattenders in the Capital Region of Denmark. Here, we report on the effectiveness of different communication platforms used in the pilot with suggestions for strategies prior to a full-implementation. Moreover, an innovative approach using self-sampling brushes with unique radio frequency identification chips allowed for unprecedented levels patient identification safety. Nonattenders from the capital region of Denmark were identified via the organized national invitation module. Screening history was obtained via the nationwide pathology registry. Twenty-four thousand women were invited, and as an alternative to the regular communication platforms (letter and phone), women could request a home test via a mobile-friendly webpage. Instruction material and video-animation in several languages were made available online. Chi-square test was used to test differences. Out of all invited, 31.7% requested a home test, and 20% returned it to the laboratory. In addition, 10% were screened at the physician after receiving the invitation. Stratified by screening history, long-term unscreened women were less likely to participate than intermittently screened women (28% vs. 16%, p < 0.001). Of all contacts received, 64% (63-65) came via letter, and 31% (95CI: 30-32%) via webpage/mobile-app. Self-sampling was well-accepted among nonattenders. Adopting modern technology-based platforms into the current organized screening program would serve as a convenient communication method between health authority and citizens, allowing easy access for the citizen and reducing the work load in administrating self-sampling approaches. © 2017 The Authors International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.

  18. Isolation of lymphotropic baboon herpesvirus (HVP) from oral swabs of hamadryas baboons of the Sukhumi monkey colony.

    Science.gov (United States)

    Agrba, V Z; Lapin, B A; Timanovskaya, V V; Dzhachvliany, M C; Kokosha, L V; Chuvirov, G N; Djatchenko, A G

    1980-01-01

    Ways of lymphotropic baboon herpesvirus (HVP) secretion and its excretion into the environment were investigated. Oral swabs and feces from the Sukhumi main stock hamadryas baboons characterized by a high risk for malignant lymphoma and the baboon stock living in isolation in the forest were used as materials for the investigations. Macaque groups of the Sukhumi stock were used as controls. It could be shown that the HVP was resistent in the oral cavity of the main stock baboons and was isolated from oral swabs of these animals both from those with malignant lymphoma and clinically healthy individuals. No virus was isolated from feces of these animals. The virus could not be isolated from oral swabs of the isolated baboon stock and macaques.

  19. Investigation of synthesized Be-bearing silicate glass as laboratory reference sample at X-ray electron probe microanalysis of silicates

    Science.gov (United States)

    Belozerova, Olga Yu.; Mikhailov, Mikhail A.; Demina, Tamara V.

    2017-01-01

    The article discusses estimates of the stability and homogeneity in Be-Mg-Al-silicate glass produced by the authors and its applicability as a laboratory reference sample for X-ray electron probe microanalysis (EPMA) of Be-bearing silicate matters: crystals and quenching melt (glasses), silicates and oxides. The results were obtained using Superprobe-733 and Superprobe JXA-8200 (JEOL Ltd, Japan) devices. The sample homogeneity was studied on macro (10-100 μm) and micro (1-10 μm) levels and was evaluated by the scheme of dispersion analysis. The applicability of Be-bearing silicate glass as a reference sample for Mg, Al, Si determinations was tested on the international certified reference glasses and laboratory reference samples of minerals with a known composition. The obtained experimental metrological characteristics correspond to the "applied geochemistry" type of analysis (second category) and suggest that Be-bearing silicate glass is appropriate as a laboratory reference sample for EPMA of Be-bearing silicate matters, silicates and oxides. Using Be-Mg-Al-silicate glass as a reference sample we obtained satisfactory data on the composition of both some minerals including cordierite and beryllium cordierite, beryllium indialite, beryl and metastable phases (chrysoberyl, compounds with structure of β-quartz and petalite).

  20. Sample pretreatment optimization for the analysis of short chain chlorinated paraffins in soil with gas chromatography-electron capture negative ion-mass spectrometry.

    Science.gov (United States)

    Chen, Laiguo; Huang, Yumei; Han, Shuang; Feng, Yongbin; Jiang, Guo; Tang, Caiming; Ye, Zhixiang; Zhan, Wei; Liu, Ming; Zhang, Sukun

    2013-01-25

    Accurately quantifying short chain chlorinated paraffins (SCCPs) in soil samples with gas chromatograph coupled with electron capture negative ionization mass spectrometry (GC-ECNI-MS) is difficult because many other polychlorinated pollutants are present in the sample matrices. These pollutants (e.g., polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and toxaphene) can cause serious interferences during SCCPs analysis with GC-MS. Four main columns packed with different adsorbents, including silica gel, Florisil and alumina, were investigated in this study to determine their performance for separating interfering pollutants from SCCPs. These experimental results suggest that the optimum cleanup procedure uses a silica gel column and a multilayer silica gel-Florisil composite column. This procedure completely separated 22 PCB congeners, 23 OCPs and three toxaphene congeners from SCCPs. However, p,p'-DDD, cis-nonachlor and o,p'-DDD were not completely removed and only 53% of the total toxaphene was removed. This optimized method was successfully and effectively applied for removing interfering pollutants from real soil samples. SCCPs in 17 soil samples from different land use areas within a suburban region were analyzed with the established method. The concentrations of SCCPs in these samples were between 7 and 541 ng g(-1) (mean: 84 ng g(-1)). Similar homologue SCCPs patterns were observed between the soil samples collected from different land use areas. In addition, lower chlorinated (Cl(6/7)) C(10)- and C(11)- SCCPs were the dominant congeners. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Viral load of equine herpesviruses 2 and 5 in nasal swabs of actively racing Standardbred trotters: Temporal relationship of shedding to clinical findings and poor performance.

    Science.gov (United States)

    Back, Helena; Ullman, Karin; Treiberg Berndtsson, Louise; Riihimäki, Miia; Penell, Johanna; Ståhl, Karl; Valarcher, Jean-François; Pringle, John

    2015-09-30

    The equine gamma herpesviruses 2 and 5 (EHV-2 and -5) have frequently been observed in the equine population and until recently presumed low to nonpathogenic. However, recent reports linking presence of equine gamma herpesviruses with clinical signs of mild to severe lung disease, suggest that the role of these viruses in respiratory disease and poor performance syndrome is still unclear. Moreover, baseline data regarding the temporal pattern of shedding of EHV-2 and EHV-5 within stables and within individual actively racing horses have been lacking. In a prospective longitudinal study, we followed elite racing Standardbred trotters at monthly intervals for 13 months, to investigate whether the amount of EHV-2 and EHV-5 shedded in nasal secretions varied over time within and between individual horses. Sixty-six elite horses were investigated by analyzing nasal swabs and serum samples, a health check and evaluation of athletic performance monthly during the study period. Nasal swabs were analyzed with two newly developed qPCR assays for EHV-2 and EHV-5, respectively. Of 663 samples, 197 (30%) were positive for EHV-2 and 492 (74%) positive for EHV-5. Furthermore, 176 (27%) of the samples were positive for both EHV-2 and EHV-5 simultaneously. There was considerable variation in the amount and frequency of shedding of EHV-2 and EHV-5 within and between individual horses. Viral load varied seasonally, but neither EHV-2 nor EHV-5 viral peaks were associated with clinical respiratory disease and/or poor performance in racing Standardbred trotters. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

    OpenAIRE

    May, Lisa A.; Higgins, Julie L.; Woodley, Cheryl M.

    2011-01-01

    The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activit...

  3. Numerical model for surge and swab pressures on wells with cross-section variation

    Energy Technology Data Exchange (ETDEWEB)

    Fedevjcyk, Joao Victor; Junqueira, Silvio Luiz de Mello; Negrao, Cezar Otaviano Ribeiro [UTFPR - Federal University of Technology - Parana - Curitiba, PR (Brazil)], e-mails: silvio@utfpr.edu.br, negrao@utfpr.edu.br

    2010-07-01

    Drilling is one of the most complex steps in petroleum exploration. The process is accomplished by rotating a drill bit to compress the rock formation. During drilling, a fluid is pumped into the well to lubricate and cool down the drill bit, to clean up the well, to avoid the formation fluid influx to the well and also to stabilize the borehole walls. Fluid circulation, however, can be interrupted for maintenance reasons and the drill pipe can be moved to remove the drill bit. The downward or upward movement of the drill pipe displaces the fluid within the well causing either under pressure (swab) or over pressure (surge), respectively. If the pressure at the well bore overcomes the formation fracture pressure, a loss of circulation can take place. On the other way round, the upward movement may reduce the pressure below the pore pressure and an inflow of fluid to the well (kick) can occur. An uncontrolled kick may cause a blowout with serious damages. The transient flow induced by the axial movement of the drill pipe is responsible for the pressure changes at the well bore. Nevertheless, the well bore cross section variation may modify the pressure change within the pipe. In this paper, the effects of diameter variation of the drilling well on the surge and swab pressures are investigated. The equations that represent the phenomenon (mass and momentum conservation) are discretized by the finite volume method. Despite its non-Newtonian properties, the fluid is considered Newtonian in this first work. The drill pipe is considered closed and the flow is assumed as single-phased, one-dimensional, isothermal, laminar, compressible and transient. A sensitivity analysis of the flow parameters is carried out. The cross-section changes cause the reflection of the pressure wave, and consequently pressure oscillations. (author)

  4. Marker-free method for accurate alignment between correlated light, cryo-light, and electron cryo-microscopy data using sample support features.

    Science.gov (United States)

    Anderson, Karen L; Page, Christopher; Swift, Mark F; Hanein, Dorit; Volkmann, Niels

    2017-11-04

    Combining fluorescence microscopy with electron cryo-tomography allows, in principle, spatial localization of tagged macromolecular assemblies and structural features within the cellular environment. To allow precise localization and scale integration between the two disparate imaging modalities, accurate alignment procedures are needed. Here, we describe a marker-free method for aligning images from light or cryo-light fluorescence microscopy and from electron cryo-microscopy that takes advantage of sample support features, namely the holes in the carbon film. We find that the accuracy of this method, as judged by prediction errors of the hole center coordinates, is better than 100 nm. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Electronic states of model hydrocarbon chromophores investigated by Synchrotron Radiation Linear Dichroism (SRLD) spectroscopy on aligned samples

    DEFF Research Database (Denmark)

    Nguyen, Duy Duc; Hoffmann, Søren Vrønning; Jones, Nykola

    2010-01-01

    Conventional UV-VIS absorption spectroscopy provides information on transition energies and intensities. Linear dichroism (LD) spectroscopy on aligned molecular samples yields additional information on transition moment directions, thereby frequently leading to resolution of otherwise overlapping...... for four hydrocarbons which are of interest as model compounds for molecular wires and switches: 1)  Diphenylethyne (tolane, DPA) 2)  1,4-Bis(phenylethynyl)benzene (BPEB) 3)  (E)-1,2-Diphenylethene (stilbene, DPE) 4)  (E,E’)-1,4-Diphenyl-1,3-butadiene (DPB)...

  6. Determination of toxic elements in plastics from waste electrical and electronic equipment by slurry sampling electrothermal atomic absorption spectrometry.

    Science.gov (United States)

    Santos, Mirian C; Nóbrega, Joaquim A; Baccan, Nivaldo; Cadore, Solange

    2010-06-15

    Cadmium, chromium, lead and antimony were determined in slurries prepared using pulverized samples of personal computers and mobile phones dispersed in dimethylformamide medium. Determinations were carried out by electrothermal atomic absorption spectrometry (ETAAS) using a graphite furnace atomic absorption spectrometer. The optimization of the experimental conditions (chemical modifier, pyrolysis time, pyrolysis temperature and atomization temperatures) was accomplished by evaluating pyrolysis and atomization curves. Optimization was also used to determine the temperatures corresponding to the best sensitivities and the lowest background signals. The pyrolysis temperatures were fixed at 600 degrees C (for Cd), 700 degrees C (for Pb), 1100 degrees C (for Sb), and 1200 degrees C (for Cr); atomization temperatures were established as 1400 degrees C (for Cd), 1300 degrees C (for Pb), 1900 degrees C (for Sb), and 2300 degrees C (for Cr), and the chemical modifier (50microg NH(4)H(2)PO(4)+3microg Mg(NO(3))(2) was used for Cd and Pb while 5microg Pd+3microg Mg(NO(3))(2) was used for Sb). The use of a chemical modifier for Cr determination was not necessary. The characteristic masses were 1.9pg for Cd, 32.3pg for Pb, 54.1pg for Sb, and 9.1pg for Cr. Calibration was performed using standard additions in a range of 5-20microgL(-1) for Cd, 5-30microgL(-1) for Cr, 12.5-50microgL(-1) for Pb, and 25-100microgL(-1) for Sb with linear correlation coefficients higher than 0.99. Limits of detection were 0.9, 1.4, 6.8, and 2.9microgL(-1) for Cd, Pb, Sb, and Cr, respectively. The results indicate that recoveries for all metals agreed at a 95% confidence level when a paired t-test was applied and presented good precision. The accuracy of the proposed method was evaluated by addition-recovery experiments, showing results in the 96-112% range, and also by comparison of the results using Student's t-test with another method developed using ETAAS for digested samples. Analyte

  7. Prevalence of population smoking cessation by electronic cigarette use status in a national sample of recent smokers.

    Science.gov (United States)

    Giovenco, Daniel P; Delnevo, Cristine D

    2018-01-01

    Amid decreasing rates of cigarette smoking and a rise in e-cigarette use, there is a need to understand population patterns of use to inform tobacco control efforts and evaluate whether e-cigarettes may play a role in tobacco harm reduction. This study merged data from the 2014 and 2015 National Health Interview Survey (NHIS) and restricted the sample to recent smokers [i.e., current smokers and former smokers who quit in 2010 or later (n=15,532)]. Log-binomial regression estimated adjusted prevalence ratios (aPR) for being quit by e-cigarette use status (i.e., daily, some day, former trier, never). All analyses controlled for factors traditionally correlated with smoking cessation. A quarter of the sample (25.2%) were former smokers. The prevalence of being quit was significantly higher among daily e-cigarette users compared to those who had never used e-cigarettes [52.2% vs. 28.2%, aPR: 3.15 (2.66, 3.73)]. Those who used e-cigarettes on some days were least likely to be former smokers (12.1%). These relationships held even after accounting for making a quit attempt and use of other tobacco products. Among those with a recent history of smoking, daily e-cigarette use was the strongest correlate of being quit at the time of the survey, suggesting that some smokers may have quit with frequent e-cigarette use or are using the products regularly to prevent smoking relapse. However, the low prevalence of cessation among infrequent e-cigarette users highlights the need to better understand this subgroup, including the individual factors and/or product characteristics that may inhibit cessation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. In-focus electrostatic Zach phase plate imaging for transmission electron microscopy with tunable phase contrast of frozen hydrated biological samples.

    Science.gov (United States)

    Frindt, Nicole; Oster, Marco; Hettler, Simon; Gamm, Björn; Dieterle, Levin; Kowalsky, Wolfgang; Gerthsen, Dagmar; Schröder, Rasmus R

    2014-02-01

    Transmission electron microscopy (TEM) images of beam sensitive weak-phase objects such as biological cryo samples usually show a very low signal-to-noise ratio. These samples have almost no amplitude contrast and instead structural information is mainly encoded in the phase contrast. To increase the sample contrast in the image, especially for low spatial frequencies, the use of phase plates for close to focus phase contrast enhancement in TEM has long been discussed. Electrostatic phase plates are favorable in particular, as their tunable potential will allow an optimal phase shift adjustment and higher resolution than film phase plates as they avoid additional scattering events in matter. Here we show the first realization of close to focus phase contrast images of actin filament cryo samples acquired using an electrostatic Zach phase plate. Both positive and negative phase contrast is shown, which is obtained by applying appropriate potentials to the phase plate. The dependence of phase contrast improvement on sample orientation with respect to the phase plate is demonstrated and single-sideband artifacts are discussed. Additionally, possibilities to reduce contamination and charging effects of the phase plate are shown.

  9. Development of a sampling method for carbonyl compounds released due to the use of electronic cigarettes and quantitation of their conversion from liquid to aerosol.

    Science.gov (United States)

    Jo, Sang-Hee; Kim, Ki-Hyun

    2016-01-15

    In this study, an experimental method for the collection and analysis of carbonyl compounds (CCs) released due to the use of electronic cigarettes (e-cigarettes or ECs) was developed and validated through a series of laboratory experiments. As part of this work, the conversion of CCs from a refill solution (e-solution) to aerosol also was investigated based on mass change tracking (MCT) approach. Aerosol samples generated from an e-cigarette were collected manually using 2,4-dinitrophenylhydrazine (DNPH) cartridges at a constant sampling (puffing) velocity of 1 L min(-1) with the following puff conditions: puff duration (2s), interpuff interval (10s), and puff number (5, 10, and 15 times). The MCT approach allowed us to improve the sampling of CCs through critical evaluation of the puff conditions in relation to the consumed quantities of refill solution. The emission concentrations of CCs remained constant when e-cigarettes were sampled at or above 10 puff. Upon aerosolization, the concentrations of formaldehyde and acetaldehyde increased 6.23- and 58.4-fold, respectively, relative to their concentrations in e-solution. Furthermore, a number of CCs were found to be present in the aerosol samples which were not detected in the initial e-solution (e.g., acetone, butyraldehyde, and o-tolualdehyde). Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Procedure for the determination of uranium on cellulose air-sampling filters by photon-electron-rejecting-alpha-liquid-scintillation spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    McDowell, W.J.; Case, G.N.

    1986-08-01

    A procedure is described for obtaining from cellulose air-sampling filters the total uranium content whether it be in the form of metal, oxide, tetrafluoride or most other salts of uranium. It is demonstrated that the uranium content can be accurately assayed by low-temperature ashing of the filter paper, dissolving the ash in a mixed nitrate-sulfate system, extracting the uranium selectively into a scintillator containing a high-molecular-weight amine sulfate, and counting the extract using a Photon-Electron-Rejecting-Alpha-Liquid-Scintillation (PERALS) spectrometer. 2 refs., 4 figs., 2 tabs.

  11. Lacrimal sac dacryoliths (86 samples): chemical and mineralogic analyses.

    Science.gov (United States)

    Komínek, Pavel; Doškářová, Sárka; Svagera, Zdeněk; Lach, Karel; Cervenka, Stanislav; Zeleník, Karol; Matoušek, Petr

    2014-03-01

    Because dacryoliths occur at low frequency, few studies have focused on their composition. We aimed to present findings from morphological, chemical, and mineralogic analysis of 86 dacryoliths. We studied 86 dacryoliths obtained during 832 dacryocystorhinostomies (DCR) performed for postsaccal obstruction. We examined the samples with atomic infrared spectrometry (80 samples), amino acid analysis (17 samples), scanning electron microscopy, and an electron microprobe with an energy dispersive detector (seven samples). Dacryoliths were found in 86/832 DCRs (10.3 %), mostly in patients with primary acquired nasolacrimal duct obstruction. All the dacryoliths were soft, composed of organic material, including proteins and mucoproteins, with approximately 20 % amino acid content. There were no "hard" dacryoliths composed of calcium phosphate. The stones were composed of lobes and lobules built on an amorphous core material with small cavities, probably as a result of various chemical processes that produced a gaseous product. The most frequent elements found in inorganic inclusions were silicon, magnesium, sulfur, potassium, calcium, sodium, and chlorine. Also, some particles had high contents of bismuth, titanium, iron, and organic fibers. The fibers found in the core of dacryoliths suggested a potential origin from cotton swabs used in cosmetics. Dacryoliths are composed almost exclusively of organic material, including proteins and mucoproteins, with approximately 20 % amino acid content.

  12. Advantages and Limitations of Direct PCR Amplification of Bacterial 16S-rDNA from Resected Heart Tissue or Swabs Followed by Direct Sequencing for Diagnosing Infective Endocarditis: A Retrospective Analysis in the Routine Clinical Setting

    Directory of Open Access Journals (Sweden)

    Daniela Maneg

    2016-01-01

    Full Text Available Infective endocarditis (IE is a life-threatening disease that is associated with high morbidity and mortality. Its long-term prognosis strongly depends on a timely and optimized antibiotic treatment. Therefore, identification of the causative pathogen is crucial and currently based on blood cultures followed by characterization and susceptibility testing of the isolate. However, antibiotic treatment starting prior to blood sampling or IE caused by fastidious or intracellular microorganisms may cause negative culture results. Here we investigate the additional diagnostic value of broad-range PCR in combination with direct sequencing on resected heart tissue or swabs in patients with tissue or swab culture-negative IE in a routine clinical setting. Sensitivity, specificity, and positive and negative predictive values of broad-range PCR from diagnostic material in our patients were 33.3%, 76.9%, 90.9%, and 14.3%, respectively. We identified a total of 20 patients (21.5% with tissue or culture-negative IE who profited by the additional application of broad-range PCR. We conclude that broad-range PCR on resected heart tissue or swabs is an important complementary diagnostic approach. It should be seen as an indispensable new tool for both the therapeutic and diagnostic management of culture-negative IE and we thus propose its possible inclusion in Duke’s diagnostic classification scheme.

  13. Comparison of SVM, RF and ELM on an Electronic Nose for the Intelligent Evaluation of Paraffin Samples

    Directory of Open Access Journals (Sweden)

    Hong Men

    2018-01-01

    Full Text Available Paraffin odor intensity is an important quality indicator when a paraffin inspection is performed. Currently, paraffin odor level assessment is mainly dependent on an artificial sensory evaluation. In this paper, we developed a paraffin odor analysis system to classify and grade four kinds of paraffin samples. The original feature set was optimized using Principal Component Analysis (PCA and Partial Least Squares (PLS. Support Vector Machine (SVM, Random Forest (RF, and Extreme Learning Machine (ELM were applied to three different feature data sets for classification and level assessment of paraffin. For classification, the model based on SVM, with an accuracy rate of 100%, was superior to that based on RF, with an accuracy rate of 98.33–100%, and ELM, with an accuracy rate of 98.01–100%. For level assessment, the R2 related to the training set was above 0.97 and the R2 related to the test set was above 0.87. Through comprehensive comparison, the generalization of the model based on ELM was superior to those based on SVM and RF. The scoring errors for the three models were 0.0016–0.3494, lower than the error of 0.5–1.0 measured by industry standard experts, meaning these methods have a higher prediction accuracy for scoring paraffin level.

  14. Comparison of SVM, RF and ELM on an Electronic Nose for the Intelligent Evaluation of Paraffin Samples

    Science.gov (United States)

    Men, Hong; Fu, Songlin; Yang, Jialin; Cheng, Meiqi; Shi, Yan

    2018-01-01

    Paraffin odor intensity is an important quality indicator when a paraffin inspection is performed. Currently, paraffin odor level assessment is mainly dependent on an artificial sensory evaluation. In this paper, we developed a paraffin odor analysis system to classify and grade four kinds of paraffin samples. The original feature set was optimized using Principal Component Analysis (PCA) and Partial Least Squares (PLS). Support Vector Machine (SVM), Random Forest (RF), and Extreme Learning Machine (ELM) were applied to three different feature data sets for classification and level assessment of paraffin. For classification, the model based on SVM, with an accuracy rate of 100%, was superior to that based on RF, with an accuracy rate of 98.33–100%, and ELM, with an accuracy rate of 98.01–100%. For level assessment, the R2 related to the training set was above 0.97 and the R2 related to the test set was above 0.87. Through comprehensive comparison, the generalization of the model based on ELM was superior to those based on SVM and RF. The scoring errors for the three models were 0.0016–0.3494, lower than the error of 0.5–1.0 measured by industry standard experts, meaning these methods have a higher prediction accuracy for scoring paraffin level. PMID:29346328

  15. To tilt or not to tilt: Correction of the distortion caused by inclined sample surfaces in low-energy electron diffraction

    Energy Technology Data Exchange (ETDEWEB)

    Sojka, Falko, E-mail: falko.sojka@uni-jena.de [Friedrich Schiller University Jena, Institute of Solid State Physics, Helmholtzweg 5, 07743 Jena (Germany); Meissner, Matthias; Zwick, Christian; Forker, Roman [Friedrich Schiller University Jena, Institute of Solid State Physics, Helmholtzweg 5, 07743 Jena (Germany); Vyshnepolsky, Michael; Klein, Claudius; Horn-von Hoegen, Michael [University of Duisburg-Essen, Department of Physics, Lotharstr. 1, 47057 Duisburg (Germany); Fritz, Torsten, E-mail: torsten.fritz@uni-jena.de [Friedrich Schiller University Jena, Institute of Solid State Physics, Helmholtzweg 5, 07743 Jena (Germany)

    2013-10-15

    Low-energy electron diffraction (LEED) is a widely employed technique for the structural characterization of crystalline surfaces and epitaxial adsorbates. For technical reasons the accessible reciprocal space is limited at a given primary electron energy E. This limitation may be overcome by sweeping E to observe higher diffraction orders decisively enhancing the quantitative examination. Yet, in many cases, such as molecular films with rather large unit cells, the adsorbate reflexes become less pronounced at energies high enough to observe substrate reflexes. One possibility to overcome this problem is an intentional inclination of the sample surface during the measurement at the expense of the quantitative interpretability of then severely distorted diffraction patterns. Here, we introduce a correction method for the axially symmetric distortion in LEED images of tilted samples. We provide experimental confirmation for micro-channel plate LEED and spot-profile analysis LEED instruments using the (7×7) reconstructed surface of a Si(111) single crystal as a reference sample. Finally, we demonstrate that the correction of this distortion considerably improves the quantitative analysis of diffraction patterns of adsorbates since substrate and adsorbate reflexes can be evaluated simultaneously. As an illustrative example we have chosen an epitaxial monolayer of 3,4,9,10-perylenetetracarboxylic dianhydride on Ag(111) that is known to form a commensurate superstructure. - Highlights: • We introduce a method to correct distortions in LEED patterns of tilted surfaces. • Higher diffraction orders unobservable at higher beam energies can be evaluated. • Our procedure makes LEED patterns of tilted samples quantitatively analyzable. • Experimental confirmation with SPA-LEED and MCP-LEED is presented. • The method is applied to PTCDA on Ag(111) confirming earlier literature values.

  16. Rapid Detection and Classification of Salmonella enterica Shedding in Feedlot Cattle Utilizing the Roka Bioscience Atlas Salmonella Detection Assay for the Analysis of Rectoanal Mucosal Swabs.

    Science.gov (United States)

    Chaney, W Evan; Agga, Getahun E; Nguyen, Scott V; Arthur, Terrance M; Bosilevac, Joseph M; Dreyling, Erin; Rishi, Anantharama; Brichta-Harhay, Dayna

    2017-10-01

    With an increasing focus on preharvest food safety, rapid methods are required for the detection and quantification of foodborne pathogens such as Salmonella enterica in beef cattle. We validated the Atlas Salmonella Detection Assay (SEN), a nucleic acid amplification technology that targets Salmonella rRNA, for the qualitative detection of S. enterica with sample enrichment using immunomagnetic separation as a reference test, and we further evaluated its accuracy to predict pathogen load using SEN signal-to-cutoff (SCO) values from unenriched samples to classify animals as high or nonhigh shedders. Rectoanal mucosal swabs (RAMS) were collected from 238 beef cattle from five cohorts located in the Midwest or southern High Plains of the United States between July 2015 and April 2016. Unenriched RAMS samples were used for the enumeration and SEN SCO analyses. Enriched samples were tested using SEN and immunomagnetic separation methods for the detection of Salmonella. The SEN method was 100% sensitive and specific for the detection of Salmonella from the enriched RAMS samples. A SEN SCO value of 8, with a sensitivity of 93.5% and specificity of 94.3%, was found to be an optimum cutoff value for classifying animals as high or nonhigh shedders from the unenriched RAMS samples. The SEN assay is a rapid and reliable method for the qualitative detection and categorization of the shedding load of Salmonella from RAMS in feedlot cattle.

  17. Electronic cigarette use in the European Union: analysis of a representative sample of 27 460 Europeans from 28 countries.

    Science.gov (United States)

    Farsalinos, Konstantinos E; Poulas, Konstantinos; Voudris, Vassilis; Le Houezec, Jacques

    2016-11-01

    To assess prevalence of electronic cigarette (e-cigarette) use, reported changes in smoking status due to e-cigarette use and correlates of e-cigarette use in the European Union (EU) member states in 2014. Cross-sectional survey of EU citizens representative of the population (Special Eurobarometer 429). All 28 Member States of the EU. A total of 27 460 EU citizens aged ≥ 15 years (after excluding those who responded 'Do not know' to the questions about smoking status and e-cigarette use). Descriptive analysis [%, 95% confidence interval (CI)] of e-cigarette use prevalence (current use, past use and past experimentation) according to smoking status, self-reported changes in smoking status according to patterns of e-cigarette use and logistic regression analysis to examine correlates of e-cigarette use, especially socio-demographic factors and smoking status. Ever e-cigarette use was reported by 31.1% (95% CI = 30.0-32.2%) of current smokers, 10.8% (95% CI = 10.0-11.7%) of former smokers and 2.3% (95% CI = 2.1-2.6%) of never smokers. Past experimentation [7.2% (95% CI = 6.9-7.5%)] was more common than current [1.8% (95% CI = 1.6-1.9%)] and past use [2.6% (95% CI = 2.4-2.8%)]. Extrapolated to the whole population, approximately 48.5 million EU citizens were ever e-cigarette users, with 76.8% using nicotine-containing e-cigarettes. An estimated 6.1 and 9.2 million EU citizens had quit and reduced smoking with the help of e-cigarettes, respectively. Initiation with e-cigarettes was reported by 0.8% (95% CI = 0.6-0.9%) of participants who reported ever use of any tobacco-related product. Only 1.3% (95% CI = 1.1-1.5%) of never smokers used nicotine-containing e-cigarettes, with 0.09% (95% CI = 0.04-0.14%) reporting daily nicotine use. Smoking cessation with the help of e-cigarettes was reported by 35.1% (95% CI = 30.7-39.5%) of current e-cigarette users, while a further 32.2% (95% CI = 29.9-36.5%) reported smoking reduction. Being current

  18. Development of a questionnaire for assessing dependence on electronic cigarettes among a large sample of ex-smoking E-cigarette users.

    Science.gov (United States)

    Foulds, Jonathan; Veldheer, Susan; Yingst, Jessica; Hrabovsky, Shari; Wilson, Stephen J; Nichols, Travis T; Eissenberg, Thomas

    2015-02-01

    Electronic cigarettes (e-cigs) are becoming increasingly popular, but little is known about their dependence potential. This study aimed to assess ratings of dependence on electronic cigarettes and retrospectively compare them with rated dependence on tobacco cigarettes among a large sample of ex-smokers who switched to e-cigs. A total of 3,609 current users of e-cigs who were ex-cigarette smokers completed a 158-item online survey about their e-cig use, including 10 items designed to assess their previous dependence on cigarettes and 10 almost identical items, worded to assess their current dependence on e-cigs (range 0-20). Scores on the 10-item Penn State (PS) Cigarette Dependence Index were significantly higher than on the comparable PS Electronic Cigarette Dependence Index (14.5 vs. 8.1, p e-cigarette users reported being less dependent on e-cigarettes than they retrospectively reported having been dependent on cigarettes prior to switching. E-cig dependence appears to vary by product characteristics and liquid nicotine concentration, and it may increase over time. © The Author 2014. Published by Oxford University Press on behalf of the Society for Research on Nicotine and Tobacco. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Measuring PM2.5, Ultrafine Particles, Nicotine Air and Wipe Samples Following the Use of Electronic Cigarettes.

    Science.gov (United States)

    Melstrom, Paul; Koszowski, Bartosz; Thanner, Meridith Hill; Hoh, Eunha; King, Brian; Bunnell, Rebecca; McAfee, Tim

    2017-09-01

    Few studies have examined the extent of inhalation or dermal contact among bystanders following short-term, secondhand e-cigarette exposure. Measure PM2.5 (particles cigarette exposure. E-cigarettes were used ad libitum by three experienced users for 2 hours during two separate sessions (disposable e-cigarettes, then tank-style e-cigarettes, or "tanks") in a 1858 ft3 room. We recorded: uncorrected PM2.5 (using SidePak); UF (using P-Trak); air nicotine concentrations (using air samplers; SKC XAD-4 canisters); ambient air exchange rate (using an air capture hood). Wipe samples were taken by wiping 100 cm2 room surfaces pre- and post- both sessions, and clean cloth wipes were worn during the exposure and collected at the end. Uncorrected PM2.5 and UF were higher (p cigarette use can produce: elevated PM2.5; elevated UF; nicotine in the air; and accumulation of nicotine on surfaces and clothing. Short-term indoor e-cigarette use produced accumulation of nicotine on surfaces and clothing, which could lead to dermal exposure to nicotine. Short-term e-cigarette use produced elevated PM2.5 and ultrafine particles, which could lead to secondhand inhalation of these particles and any chemicals associated with them by bystanders. We measured significant differences in PM2.5 and ultrafine particles between disposable e-cigarettes and tank-style e-cigarettes, suggesting a difference in the exposure profiles of e-cigarette products.

  20. [Influence of distinct criteria for selecting patients for swabbing on estimation of the effectiveness of the influenza vaccine].

    Science.gov (United States)

    Martínez-Baz, Iván; Guevara, Marcela; Elía, Fernando; Ezpeleta, Carmen; Fernández Alonso, Mirian; Castilla, Jesús

    2014-01-01

    To estimate the effectiveness of the influenza vaccine under different criteria for selecting patients for swabbing. A case-control study was performed of laboratory-confirmed cases (n=909) and negative controls for influenza (n=732) in the 2010-2011 to 2012-2013 seasons in Navarre (Spain). The adjusted vaccine effectiveness was estimated by including all swabs from patients with influenza-like-illness and selecting only the first two cases per physician and week. The first two patients per physician and week were less frequently vaccinated against influenza (7.9% vs. 12.5%, p=0.021) and less often received confirmation of influenza (53.6% vs. 66.4%, p <0.001) than subsequent patients. These differences decreased after adjustment for covariates. The effectiveness of the influenza vaccine was 49% (95% CI: 23-66%) when all swabs were included and was 55% (95% CI: 27-72%) when we selected the first two swabs per week and physician. The selection of the first two patients per physician and week may bias assessment of the effectiveness of the influenza vaccine, although this bias was small in the seasons analyzed. Copyright © 2013 SESPAS. Published by Elsevier Espana. All rights reserved.

  1. Evaluation of penicillin G residues by kidney inhibition swab tests in sow body fluids and tissues following intramuscular injection

    Science.gov (United States)

    In 2011, the USDA-Food Safety and Inspection Service (FSIS) changed the method used for screening swine tissues for antimicrobial residues from the Fast Antimicrobial Screen Test to the Kidney Inhibition Swab (KIS(TM)). Here, we describe the use of KIS(TM) test for the detection of penicillin G res...

  2. Optimisation of the antibiotic guidelines in The Netherlands. VII. SWAB guidelines for antimicrobial therapy in adult patients with infectious endocarditis

    NARCIS (Netherlands)

    Verhagen, D. W. M.; van der Feltz, M.; Plokker, H. W. M.; Buiting, A. G. M.; Tjoeng, M. M.; van der Meer, J. T. M.

    2003-01-01

    The Working Party on Antibiotic Policy (Dutch acronym is SWAB) is a Dutch organisation that develops guidelines for in-hospital antimicrobial therapy of bacterial infectious diseases. This present guideline describes the antimicrobial treatment for adult patients with infective endocarditis. The

  3. [Optimizing antibiotics policy in the Netherlands. VI. SWAB advice: no selective decontamination of intensive care patients on mechanical ventilation

    NARCIS (Netherlands)

    Bonten, M.J.; Kullberg, B.J.; Filius, P.M.

    2001-01-01

    The Working Party on Antibiotic Policy (Dutch acronym is SWAB) has issued a guideline in which the pro and cons of the routine use of selective decontamination (SD) in patients in intensive care (IC) on mechanical ventilation are compared in order to decide whether SD is indicated. The effectiveness

  4. Evaluation of a New Environmental Sampling Protocol for Detection of Human Norovirus on Inanimate Surfaces

    Science.gov (United States)

    Lee, David; Treffiletti, Aimee; Hrsak, Mario; Shugart, Jill; Vinjé, Jan

    2015-01-01

    Inanimate surfaces are regarded as key vehicles for the spread of human norovirus during outbreaks. ISO method 15216 involves the use of cotton swabs for environmental sampling from food surfaces and fomites for the detection of norovirus genogroup I (GI) and GII. We evaluated the effects of the virus drying time (1, 8, 24, or 48 h), swab material (cotton, polyester, rayon, macrofoam, or an antistatic wipe), surface (stainless steel or a toilet seat), and area of the swabbed surface (25.8 cm2 to 645.0 cm2) on the recovery of human norovirus. Macrofoam swabs produced the highest rate of recovery of norovirus from surfaces as large as 645 cm2. The rates of recovery ranged from 2.2 to 36.0% for virus seeded on stainless-steel coupons (645.0 cm2) to 1.2 to 33.6% for toilet seat surfaces (700 cm2), with detection limits of 3.5 log10 and 4.0 log10 RNA copies. We used macrofoam swabs to collect environmental samples from several case cabins and common areas of a cruise ship where passengers had reported viral gastroenteritis symptoms. Seventeen (18.5%) of 92 samples tested positive for norovirus GII, and 4 samples could be sequenced and had identical GII.1 sequences. The viral loads of the swab samples from the cabins of the sick passengers ranged from 80 to 31,217 RNA copies, compared with 16 to 113 RNA copies for swab samples from public spaces. In conclusion, our swab protocol for norovirus may be a useful tool for outbreak investigations when no clinical samples are available to confirm the etiology. PMID:26116675

  5. Gardnerella vaginalis and Mollicute detection in rectal swabs from men who have sex with men.

    Science.gov (United States)

    Cox, Ciara; Watt, Alison P; McKenna, James P; Coyle, Peter V

    2017-06-01

    The numbers of rectal sexually transmitted infections are on the rise especially among men who have sex with men. Males from men who have sex with men population are encouraged to send a rectal swab to the laboratory for sexually transmitted infection screening at their visit to the Genitourinary Medicine Clinic. In healthy asymptomatic males, the range of pathogens tested is limited therefore other pathogens may be left untreated allowing infections to persist among sexual partners. Molecular techniques have revolutionarised sexually transmitted infection testing enabling the detection of previously difficult-to-culture pathogens in extra-genital sites and have increased the evidence base for their clinical significance. The present study tests 107 rectal swabs from men who have sex with men negative for Chlamydia trachomatis and Neisseria gonorrhoeae against quantitative polymerase chain reaction (qPCR) assays targeting five common sexually transmitted bacteria which include Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum and Gardnerella vaginalis. The pathogenic role of these five bacteria in men who have sex with men is currently unknown. Amongst the 107 patients, a positive qPCR was obtained respectively for G. vaginalis 89 (83.2%); U. urealyticum 26 (24.3%); M. hominis 26 (24.3%); M. genitalium 10 (9.3%) and U. parvum 5 (4.7%). Bacterial loads in single and co-infections were compared for each organism. G. vaginalis and M. hominis loads were significantly ( p = 0.007 and p = 0.005, respectively) higher when co-infecting with at least one other organism. Amongst co-infections, the loads of each organism were assessed to determine possible synergies. G. vaginalis and M. hominis displayed a synergistic pattern ( r = 0.51; p = 0.02) which is in keeping with a similar synergy detected previously in the vagina of women with bacterial vaginosis. This study outlines that potential significant infections are being

  6. Consensus of microbiology reporting of ear swab results to primary care clinicians in patients with otitis externa.

    Science.gov (United States)

    Geyer, M; Howell-Jones, R; Cunningham, R; McNulty, C

    2011-01-01

    Otitis externa is a ubiquitous inflammatory disease; although it arises most commonly from an infection, there is no consensus in the UK for the reporting of ear swab culture results. This study aims to review current microbiology laboratory reporting of ear swab specimens to primary care and reach an evidence-based consensus for a reporting policy. Fifty consecutive ear swab reports were reviewed from each of 12 laboratories in the South West region to determine and discuss reporting practice. The Health Protection Agency (HPA) GP Microbiology Laboratory Use Group reviewed the underlying evidence and worked towards a consensus of expert microbiology opinion for laboratory reporting of ear swab results using a modified version of the Delphi technique. A total of 487 reports from primary care were reviewed (54% female; 46% male). Cultures most commonly yielded Pseudomonas species (36%), Staphylococcus species (21%), Streptococcus species (15%) and fungi (11%). Five reporting policies were agreed: Policy 1: Common pathogens such as group A beta-haemolytic streptococci, Streptococcus pneumoniae, Staphylococcus aureus - Always reported by name with antibiotic susceptibilities. Policy 2: Pseudomonas species - Always reported, but antibiotic susceptibilities only reported in severe disease. Policy 3: Aspergillus, Candida, coliforms and Proteus species, as well as non-group A streptococci and anaerobes - Only reported if moderate numbers of colonies and it is the predominant organism present; if appropriate report antibiotic susceptibilities. Policy 4: Coagulase-negative staphylococci, diphtheroids and enterococci - Not reported by name; generic terms used and antibiotic susceptibilities not reported. Policy 5: When antibiotic susceptibilities reported these must include susceptibility to a topical antibiotic. It is suggested that laboratories should consider adopting this evidence-based reporting consensus for ear swab culture results from primary care patients with

  7. Flow injection of liquid samples to a mass spectrometer with ionization under vacuum conditions: a combined ion source for single-photon and electron impact ionization.

    Science.gov (United States)

    Schepler, Claudia; Sklorz, Martin; Passig, Johannes; Famiglini, Giorgio; Cappiello, Achille; Zimmermann, Ralf

    2013-09-01

    Electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI), and atmospheric pressure photo-ionization (APPI) are the most important techniques for the ionization of liquid samples. However, working under atmospheric pressure conditions, all these techniques involve some chemical rather than purely physical processes, and therefore, side reactions often yield to matrix-dependent ionization efficiencies. Here, a system is presented that combines both soft single-photon ionization (SPI) and hard 70 eV electron impact ionization (EI) of dissolved compounds under vacuum conditions. A quadrupole mass spectrometer was modified to enable direct EI, a technique developed by Cappiello et al. to obtain library-searchable EI mass spectra as well as soft SPI mass spectra of sample solutions. An electron beam-pumped rare gas excimer lamp working at 126 nm was used as well as a focusable vacuum UV light source for single-photon ionization. Both techniques, EI and SPI, were applied successfully for flow injection experiments providing library-matchable EI fragment mass spectra and soft SPI mass spectra, showing dominant signals for the molecular ion. Four model compounds were analyzed: hexadecane, propofol, chlorpropham, and eugenol, with detection limits in the picomolar range. This novel combination of EI and SPI promises great analytical benefits, thanks to the possibility of combining database alignment for EI data and molecular mass information provided by SPI. Possible applications for the presented ionization technology system are a matrix-effect-free detection and a rapid screening of different complex mixtures without time-consuming sample preparation or separation techniques (e.g., for analysis of reaction solutions in combinatorial chemistry) or a switchable hard (EI) and soft (SPI) MS method as detection step for liquid chromatography.

  8. Evaluation of a Self-Administered Intravaginal Swab for PCR Detection of Genitourinary Tract Infections Including Chlamydia, Gonorrhea, Trichomonas and Human Papillomavirus in Active Duty Military Women

    Science.gov (United States)

    1998-10-01

    Trichomonas bmcei subsp. rhodesiense, K02836; Trypanosoma cmzi, M97956; Toxop/asma gon- gallinae ATCC 30002, Giardia lamblia ATCC SF-741 30888, Chilomastix...Administered Intravaginal Swab for PCR Detection of Genitourinary Tract Infections Including Chlamydia, Gonorrhea, Trichomonas and Human...FUNDING NUMBERS Intravaginal Swab for PCR Detection of Genitourinary DAMD17-96-1-6309 Tract Infections Including Chlamydia, Gonorrhea, Trichomonas

  9. Isolation of thermotolerant campylobacters and C. hyointestinalis from rectal swabs of healthy pigs

    Directory of Open Access Journals (Sweden)

    Mrenoski Slavco

    2007-11-01

    Full Text Available Thermotolerant campylobacters are the most common bacterial etiological agents of human infectious gastroenteritis worldwide. The most frequent isolated species among them are Campylobacter jejuni and C. coli, and less frequent C. upsaliensis and C. lari. Also C. hyointestinalis, that not belong to the group of thermotolerant campylobacters, has been indicate as an agent of human infectious gastroenteritis. Natural reservoir of all named campylobacters is the intestinal tract of many mammals and birds, including domestic animals. In these animals, campylobacters are commonly present as commensals and their feces is considered as a prime source for environmental contamination. Unlike the human feces which is usually examined in the cases of diarrhea, thermotolerant campylobacters and C. hyointestinalis in the animal feces are generally present in a much lesser amount and the isolation very often could be unsuccessful. The aim of this study was to estimate the validity of applied procedure for isolation (and identification of thermotolerant campylobacters and C. hyointestinalis from pig rectal swabs, as a procedure for detection of healthy animal carriers.

  10. Impact of Rectal Swabs on Infectious Complications after Transrectal Prostate Biopsy.

    Science.gov (United States)

    Trujillo, Carlos Gustavo; Plata, Mauricio; Caicedo, Juan Ignacio; Cataño Cataño, Juan Guillermo; Mariño Alvarez, Angela Marcela; Castelblanco, Diana; Robledo, Daniela

    2016-01-01

    To determine the impact of rectal swabs (RSs) on infectious complications (IC) following prostate biopsy (PB). A retrospective cohort study was conducted including all patients subjected to PB between 2009 and 2013. Group B consisted of patients with a RS and group A of patients without. RS reported the presence of gram-positive or negative germs, sensitive or resistant to ciprofloxacin. Antimicrobial prophylaxis was adjusted to the result. Frequency of IC in each group was determined. Group B had 548 (47.20%) patients and group A 613 (52.80%). From group B, 250 (45.62%) of the RSs showed fluoroquinolone (FQ)-resistant germs. Forty nine (16.44%) patients with sensitive germs vs. 147 (59.51%) with resistant germs had a history of previous FQ treatment (p < 0.0001). IC were observed in 33 (5.49%) patients from group A and in 7 (1.28%) patients from group B (p < 0.0001), requiring hospitalization in 4.99 vs. 1.28%, respectively. IC and hospital admissions were reduced in 76.68 and 74.34%, respectively, following the implementation of RS. RS and targeted antibiotic prophylaxis prior to PB was associated with a significant reduction in IC and hospital admissions. Ceftriaxone could be an alternative in cases of known resistance. Past history of FQ treatment is associated with increased resistance. © 2016 S. Karger AG, Basel.

  11. ANALYSIS AND IDENTIFICATION SPIKING CHEMICAL COMPOUNDS RELATED TO CHEMICAL WEAPON CONVENTION IN UNKNOWN WATER SAMPLES USING GAS CHROMATOGRAPHY AND GAS CHROMATOGRAPHY ELECTRON IONIZATION MASS SPECTROMETRY

    Directory of Open Access Journals (Sweden)

    Harry Budiman

    2010-06-01

    Full Text Available The identification and analysis of chemical warfare agents and their degradation products is one of important component for the implementation of the convention. Nowadays, the analytical method for determination chemical warfare agent and their degradation products has been developing and improving. In order to get the sufficient analytical data as recommended by OPCW especially in Proficiency Testing, the spiking chemical compounds related to Chemical Weapon Convention in unknown water sample were determined using two different techniques such as gas chromatography and gas chromatography electron-impact ionization mass spectrometry. Neutral organic extraction, pH 11 organic extraction, cation exchanged-methylation, triethylamine/methanol-silylation were performed to extract the chemical warfare agents from the sample, before analyzing with gas chromatography. The identification of chemical warfare agents was carried out by comparing the mass spectrum of chemicals with mass spectrum reference from the OPCW Central Analytical Database (OCAD library while the retention indices calculation obtained from gas chromatography analysis was used to get the confirmation and supported data of  the chemical warfare agents. Diisopropyl methylphosphonate, 2,2-diphenyl-2-hydroacetic acid and 3-quinuclidinol were found in unknown water sample. Those chemicals were classified in schedule 2 as precursor or reactant of chemical weapons compound in schedule list of Chemical Weapon Convention.   Keywords: gas chromatography, mass spectrometry, retention indices, OCAD library, chemical warfare agents

  12. Rapid analysis of pyrethroid insecticides in aquaculture seawater samples via membrane-assisted solvent extraction coupled with gas chromatography-electron capture detection.

    Science.gov (United States)

    Shi, X Z; Song, S Q; Sun, A L; Liu, J H; Li, D X; Chen, J

    2012-01-21

    A simple, efficient, and environmentally friendly membrane-assisted solvent extraction (MASE) method for the extraction and preconcentration of six pyrethroid insecticides from aquaculture seawater samples followed by gas chromatography-electron capture detection (GC-ECD) was successfully proposed. The operating conditions for MASE, such as the extraction solvent, solvent volume, NaCl concentration, stirring rate, extraction time, and temperature, were optimized. Compared to conventional Florisil-solid phase extraction (SPE), higher extraction recoveries (85.9% to 105.9%) of three spiked levels of the six pyrethroid pesticides in aquaculture seawater were obtained using MASE, and the RSD values were lower than 7.9%. The limits of detection (LOD, signal-to-noise ratio (S/N)=3) and quantification (LOQ, S/N = 10) were in the range of 0.037-0.166 and 0.12-0.55 μg L(-1), respectively. The results demonstrate the excellent applicability of the MASE method in analyzing the six pyrethroid pesticides in aqueous samples. The proposed method exhibited a high potential for routine monitoring analysis of pyrethroid insecticides in seawater samples.

  13. Development of a non-destructive micro-analytical method for stable carbon isotope analysis of transmission electron microscope (TEM) samples

    Energy Technology Data Exchange (ETDEWEB)

    Hode, Tomas [Department of Geology, Portland State University, Portland, P.O. Box 751, OR 97201 (United States)], E-mail: hode@pdx.edu; Kristiansson, Per; Elfman, Mikael [Division of Nuclear Physics, Department of Physics, Lund Institute of Technology, Lund University, P.O. Box 118, SE-221 00 Lund (Sweden); Hugo, Richard C.; Cady, Sherry L. [Department of Geology, Portland State University, Portland, P.O. Box 751, OR 97201 (United States)

    2009-10-01

    The biogenicity of ancient morphological microfossil-like objects can be established by linking morphological (e.g. cell remnants and extracellular polymeric matrix) and chemical (e.g. isotopes, biomarkers and biominerals) evidence indicative of microorganisms or microbial activity. We have developed a non-destructive micro-analytical ion beam system capable of measuring with high spatial resolution the stable carbon isotope ratios of thin samples used for transmission electron microscopy. The technique is based on elastic scattering of alpha particles with an energy of 2.751 MeV. At this energy the {sup 13}C cross section is enhanced relative to the pure Rutherford cross section for {sup 13}C, whereas the {sup 12}C cross section is reduced relative to its pure Rutherford cross section. Here we report the initial results of this experimental approach used to characterize ultramicrotomed sections of sulfur-embedded graphite and microbial cells.

  14. A comparative study of sampling techniques for monitoring carcass contamination

    NARCIS (Netherlands)

    Snijders, J.M.A.; Janssen, M.H.W.; Gerats, G.E.; Corstiaensen, G.P.

    1984-01-01

    Four bacteriological sampling techniques i.e. the excision, double swab, agar contract and modified agar contact techniques were compared by sampling pig carcasses before and after chilling. As well as assessing the advantages and disadvantages of the techniques particular attention was paid to

  15. Detection of Actinobacillus pleuropneumoniae in cultures from nasal and tonsillar swabs of pigs by a PCR assay based on the nucleotide sequence of a dsbE-like gene.

    Science.gov (United States)

    Chiers, K; Van Overbeke, I; Donné, E; Baele, M; Ducatelle, R; De Baere, T; Haesebrouck, F

    2001-11-08

    A PCR assay for the detection of Actinobacillus pleuropneumoniae was developed based on the amplification of a dsbE-like gene. All of 157 field isolates of A. pleuropneumoniae reacted in the PCR by the amplification of a 342bp product. No reaction was observed with related bacterial species or other bacterial species isolated from pigs, except for A. lignieresii. The lower detection limit of the PCR was 10(2) CFU per PCR test tube and was not affected by the addition of 10(6) CFU Escherichia coli. The PCR was evaluated on mixed bacterial cultures from nasal and tonsillar swabs as well as suspensions of nasal conchae and tonsils obtained from specific pathogen-free (SPF) pigs, experimentally infected pigs, and pigs from farrow-to-finish herds. The results of the new PCR were compared with a PCR based on the detection of the omlA gene coding for an outer membrane protein, with a commercially available PCR (Adiavet APP, Adiagène, Saint-Brieuc, France), and with conventional culturing. No positive reactions were observed with any of the PCR methods in samples of SPF animals. In samples of the other animals, no or low significant differences between nasal swabs and suspensions as well as tonsillar swabs and suspensions were observed in any method. In general, more positive results were obtained from tonsillar samples in comparison to nasal samples. Interassay sensitivity and specificity values were assessed for each test by pair wise comparisons between assays. The agreement between tests was evaluated by calculating Cohen's kappa coefficient. From these analyses the three PCR assays showed a good agreement. The dsbE-based PCR proved to be highly sensitive (95 and 93%) and specific (82 and 74%) in comparison to the omlA-based PCR and the commercially available PCR, respectively. It was concluded that the dsbE-like gene-based PCR is a reliable diagnostic assay for demonstration of A. pleuropneumoniae. Furthermore, it was demonstrated that tonsillar swabs can be used for

  16. Collection by trained pediatricians or parents of mid-turbinate nasal flocked swabs for the detection of influenza viruses in childhood

    Directory of Open Access Journals (Sweden)

    Fossali Emilio

    2010-04-01

    Full Text Available Abstract This study evaluated the efficiency of pediatric mid-turbinate nasal flocked swabs used by parents in 203 children aged 6 months to 5 years with signs and symptoms of respiratory disease. Two nasal samples were collected from each child in a randomised sequence: one by a trained pediatrician and one by a parent. The real-time polymerase chain reaction influenza virus detection rates were similar in the samples collected using the two methods (Cohen's kappa = 0.86, as were the cycle threshold values. In comparison with the pediatrician-collected samples, the sensitivity and specificity of the parental collections were respectively 89.3% (95% confidence interval [CI]: 77.8-100% and 97.7% (95% CI: 95.5-100%, and the positive and negative predictive values were respectively 86.2% (95% CI: 73.7-95.1% and 98.2% (95% CI: 96.4-100%. The children were significantly more satisfied with the parental collections (median values ± standard deviation, 1.59 ± 0.55 vs 3.51 ± 0.36; p

  17. Utility of nasal swab and age in detecting methicillin-resistant Staphylococcus aureus in pediatric head and neck abscesses.

    Science.gov (United States)

    Bradford, Benjamin D; Macias, David; Liu, Yuan F; Inman, Jared C; Dyleski, Robin A

    2017-10-01

    To identify risk factors associated with the presence of methicillin-resistant Staphylococcus aureus (MRSA) in surgical cultures taken from incision and drainage (I&D) of head and neck abscesses in the pediatric population. Retrospective case series. All patients under 18 years of age with a head and neck abscess requiring I&D from 2009 to 2015 were reviewed. MRSA nasal swab cultures were taken from all patients upon hospitalization. Surgical cultures were obtained from all patients and correlated with MRSA nasal swab results. Univariate and multivariate logistic regression was performed, and odds ratios (ORs) along with descriptive statistics were analyzed. Of a total of 272 patients, there were 68 (25%) MRSA-positive abscesses. The majority (86.8%) of these abscesses were in children under 2 years of age. Overall, 12 (4.4%) presented with positive admission MRSA nasal swabs. Of these, 91.7% had MRSA-positive abscess cultures. Decreasing age in years showed an OR of 1.650 (P MRSA-positive abscess, with children less than 1 year old having the highest OR of 10.74 (P MRSA nasal colonization were two statistically significant risk factors for developing an MRSA abscess of the head and neck. This study demonstrates a high positive predictive value for MRSA-positive neck abscesses when nasal swab screenings were MRSA-positive (91.7%). Children under 2 years of age-especially those under 1 year of age-or those with MRSA nasal colonization can be considered a high-risk population that may benefit from empiric antibiotics against MRSA for head and neck abscesses. 4. Laryngoscope, 127:2407-2412, 2017. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

  18. Clinical performance of the Solana® Point-of-Care Trichomonas Assay from clinician-collected vaginal swabs and urine specimens from symptomatic and asymptomatic women.

    Science.gov (United States)

    Gaydos, C A; Schwebke, J; Dombrowski, J; Marrazzo, J; Coleman, J; Silver, B; Barnes, M; Crane, L; Fine, P

    2017-03-01

    Solana® (Quidel) is a new rapid (Trichomonas vaginalis (TV) DNA. The assay has two steps: 1) specimen preparation, and 2) amplification and detection using isothermal Helicase-Dependent Amplification (HDA). The objective was to demonstrate the performance of Solana for vaginal swabs and female urines based on comparison to wet mount and TV culture. Performance was also compared to the Aptima-TV assay. Urine and four clinician-collected vaginal swabs were collected. The first two were used for FDA composite reference (wet mount; InPouch TV Culture). The third swab was used for Solana. Sensitivity/specificity were based on the reference method. A specimen was considered positive if either test was positive. The fourth swab was for Aptima-TV. Vaginal swabs and urines were obtained from 501 asymptomatic and 543 symptomatic women. Prevalence of TV by was 11.5%. For swabs, Solana® demonstrated high sensitivity and specificity from asymptomatic (100%/98.9%) and symptomatic (98.6%/98.5%) women, as well as for urines from asymptomatic (98.0%/98.4%) and symptomatic (92.9%/97.9%) women, compared to the reference method. Compared to Aptima-TV, the sensitivity/specificity was 89.7%/99.0% for swabs and 100%/98.9% for urines. The Solana® assay performed well compared to the reference assays.

  19. Isolation of Chlamydia trachomatis from men with urethritis: relative value of one vs. two swabs and influence of concomitant gonococcal infection.

    Science.gov (United States)

    Singal, S S; Reichman, R C; Graman, P S; Greisberger, C; Trupei, M A; Menegus, M A

    1986-01-01

    Two successive urethral swabs were used to obtain specimens for culture of Neisseria gonorrhoeae and Chlamydia trachomatis from 136 heterosexual men with urethritis. The first swab was used to culture N. gonorrhoeae and then C. trachomatis; the second was used to culture C. trachomatis only. C. trachomatis cultures from the second swab were positive more often (30 of 31 pairs) than were cultures from the first swab (22 of 31 pairs) (P less than .05). In addition, cultures from swab 2 had greater numbers of inclusions per coverslip more frequently (23 of 31 pairs) than did cultures from the first swab (six of 31 pairs) (P = .003). Numbers of chlamydial inclusions per coverslip were lower in specimens positive for both C. trachomatis and N. gonorrhoeae than in specimens positive for C. trachomatis only (P less than .02). In addition, the presence of N. gonorrhoeae in a specimen adversely affected the quality of the McCoy cell monolayer. In 17 of 21 instances of monolayer toxicity, cultures for N. gonorrhoeae were positive (P less than .01). These results demonstrate that when specimens from men with urethritis are cultured for N. gonorrhoeae and C. trachomatis, use of a second swab will improve rates of recovery of C. trachomatis. Material present in specimens that contain N. gonorrhoeae may adversely affect rates of isolation of C. trachomatis.

  20. Diagnostic performance of rectal swab versus bulk stool specimens for the detection of rotavirus and norovirus: Implications for outbreak investigations✩

    Science.gov (United States)

    Arvelo, Wences; Hall, Aron J.; Estevez, Alejandra; Lopez, Beatriz; Gregoricus, Nicole; Vinjé, Jan; Gentsch, Jon R.; Parashar, Umesh; Lindblade, Kim A.

    2015-01-01

    Background In January of 2008, during the peak of the rotavirus season in Guatemala, a gastroenteritis outbreak with high mortality among infants was reported in Guatemala. Despite extensive efforts, the investigation was limited by the lack of bulk stool specimens collected, particularly from the more severely dehydrated or deceased children. Objectives We evaluated the diagnostic performance of rectal swab specimens compared with bulk stool for the detection of rotavirus and norovirus. Study design Patients with diarrhea (≥3 loose stools in 24 h) were enrolled through an ongoing surveillance system in Guatemala. From January through March 2009, we attempted to enroll 100 patients <5years old captured by the diarrhea surveillance, and collected paired bulk stool and rectal swabs specimens from them. Specimens were tested for norovirus using real-time reverse transcription-polymerase chain reaction and for rotavirus via enzyme immunoassay. Results We enrolled 102 patients with paired specimens; 91% of 100 paired specimens tested for rotavirus yielded concordant results positive for rotavirus with a negativity rate of 83%. Among 100 paired specimens tested for norovirus, 86% were concordant norovirus detection and the negativity rate was 85%. The diagnostic performance for rotavirus and norovirus detection did not differ significantly between the two specimen types. Conclusions Testing of properly collected fecal specimens using rectal swabs may be a viable alternative to bulk stool for detection of rotavirus and norovirus, particularly during outbreaks where collection of bulk stool may be difficult. PMID:24139675

  1. Vaginal Self-Swab Specimen Collection in a Home-Based Survey of Older Women: Methods and Applications

    Science.gov (United States)

    Hoffmann, Joscelyn N.; Lundeen, Katie; Jaszczak, Angela; McClintock, Martha K.; Jordan, Jeanne A.

    2009-01-01

    Objectives To describe the methods used for, cooperation with, assays conducted on, and applications of vaginal specimens collected by older women in their homes. Methods Community-residing women (N = 1,550), ages 57–85 years, participated in a nationally representative probability survey. Vaginal self-swab specimen collection and in-home interviews were conducted between 2005 and 2006. Specimens were analyzed for bacterial vaginosis (BV), vaginal candidiasis (VC), high-risk human papillomavirus (HR-HPV), and cytological characteristics. Field methods, consent procedures, the swab protocol, laboratory procedures, and results reporting are described. Results One thousand twenty-eight respondents (67.5% weighted) agreed to provide a vaginal specimen; 99.1% were successful. The specimen adequacy rates were BV and VC, 94.1%; HR-HPV, 99.7%; and cytology, 85.5%. The most common recorded reason for nonparticipation was a physical or health problem (38% of nonresponders). Responders were significantly more likely than nonresponders to be younger and more educated, and were more likely to report a recent pelvic examination, menopausal hormone use, and recent sexual activity. Discussion Collection of vaginal self-swab specimens from older women in a population-based study is feasible and provides novel data on microenvironmental characteristics of the female genital tract relevant to analyses of gynecologic health, sexual activity and problems, and immune and inflammatory function. PMID:19204072

  2. Sterilization monitoring by biological indicators and conventional swab test of different sterilization processes used in orthodontics: A comparative study

    Directory of Open Access Journals (Sweden)

    Shantanu Khattri

    2015-01-01

    Full Text Available Introduction: The need of effective sterilization method and their monitoring is necessary. Biological indicators are specific microorganisms with high resistance toward particular sterilization methods. Their processes include steam autoclave, dry heat sterilizer, ethylene oxide sterilizer. This article has considered various methods to monitor the effectiveness of different sterilization methods used in orthodontics. Materials and Methods: The parameters for comparison were the control and experimental instruments utilized in orthodontic treatment. The efficacy of sterilization was evaluated by comparison of bacterial growth obtained in monitoring by biological indicators and swab test method. Results: No spore growth was found when sterilization process was evaluated by biological indicators in comparison to swab test where spore growth was present. Instruments dipped in Bioclenz-G solution for 10 min showed spore growth, but no spore growth was seen in 10 h cycle. Discussion: The result of the study verifies the established effectiveness of biological indicators over conventional swab test method in monitoring various sterilization processes used in orthodontics. Bioclenz-G solution can be used as an effective cold sterilization method for sterilization. Conclusion: For evaluating the effectiveness of sterilization, biological indicators preclude the drawbacks of incomplete verification of destruction of all vegetation and inordinate delay in procurement of results as is the case with chemical indicators and lab culture, respectively.

  3. Estimation of the sensitivity of four sampling methods for Mycoplasma hyopneumoniae detection in live pigs using a Bayesian approach.

    Science.gov (United States)

    Fablet, C; Marois, C; Kobisch, M; Madec, F; Rose, N

    2010-07-14

    Four sampling techniques for Mycoplasma hyopneumoniae detection, namely nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing, were compared in naturally infected live pigs. In addition, a quantitative real-time PCR assay for M. hyopneumoniae quantification was validated with the same samples. 60 finishing pigs were randomly selected from a batch of contemporary pigs on a farm chronically affected by respiratory disorders. Each pig was submitted to nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing. Nested-PCR and real-time PCR assays were performed on all samples. A Bayesian approach was used to analyze the nested-PCR results of the four sampling methods (i.e. positive or negative) to estimate the sensitivity and specificity of each method. M. hyopneumoniae was detected by nested-PCR in at least one sample from 70% of the pigs. The most sensitive sampling methods for detecting M. hyopneumoniae in live naturally infected pigs were tracheo-bronchial swabbing and tracheo-bronchial washing, as compared to oral-pharyngeal brushing and nasal swabbing. Swabbing the nasal cavities appeared to be the least sensitive method. Significantly higher amounts of M. hyopneumoniae DNA were found at the sites of tracheo-bronchial sampling than in the nasal cavities or at the oral-pharyngeal site (p0.05). Our study indicated that tracheo-bronchial swabbing associated with real-time PCR could be an accurate diagnostic tool for assessing infection dynamics in pig herds. (c) 2009 Elsevier B.V. All rights reserved.

  4. Evaluating intra- and inter- sample variability in Electron Spin Resonance dating of fossil teeth: an example from Cuesta de la Bajada site (Spain).

    Science.gov (United States)

    Duval, Mathieu; Grün, Rainer; Shao, Qingfeng; Martin, Loïc; Arnold, Lee J.

    2017-04-01

    Over the last decades, technological improvements have progressively enabled to significantly decrease the amount of material required for dating analyses. In particular, the combined use of laser ablation (LA) with ICP-MS opened new possibilities for high resolution in situ U-series analyses of fossil teeth. With this technique it is now possible to directly visualise the spatial distribution of U and Th isotopes in dental tissues. Moreover, the combination of LA-ICP-MS with Electron Spin Resonance (ESR) enables an increased sampling resolution, and offers the possibility to produce several ages for different areas within a given fossil tooth. To test the potential of this new approach, several fossil teeth were collected from the Middle Palaeolithic site of Cuesta de la Bajada (Teruel, Spain). Each tooth was divided into several subsamples, providing thus several combined US-ESR age results per tooth. For each subsample, ESR, high-resolution laser ablation and solution ICP-MS U-series analyses were systematically performed. Relative beta dose rate contributions from the different tissues and the sediment were also adjusted using DosiVox software and compared with those derived from the standard approach. The results of this work give some interesting insight into the intra- and inter- sample variability that may exist at a given site. The consistency of the final US-ESR age estimates obtained on teeth are also evaluated by comparison with the (semi)-independent results derived from ESR and Luminescence dating of optically bleached quartz grains collected from the same excavation area.

  5. Orthogonal identification of gunshot residue with complementary detection principles of voltammetry, scanning electron microscopy, and energy-dispersive X-ray spectroscopy: sample, screen, and confirm.

    Science.gov (United States)

    O'Mahony, Aoife M; Samek, Izabela A; Sattayasamitsathit, Sirilak; Wang, Joseph

    2014-08-19

    Field-deployable voltammetric screening coupled with complementary laboratory-based analysis to confirm the presence of gunshot residue (GSR) from the hands of a subject who has handled, loaded, or discharged a firearm is described. This protocol implements the orthogonal identification of the presence of GSR utilizing square-wave stripping voltammetry (SWSV) as a rapid screening tool along with scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) to confirm the presence of the characteristic morphology and metal composition of GSR particles. This is achieved through the judicious modification of the working electrode of a carbon screen-printed electrode (CSPE) with carbon tape (used in SEM analysis) to fix and retain a sample. A comparison between a subject who has handled and loaded a firearm and a subject who has had no contact with GSR shows the significant variations in voltammetric signals and the presence or absence of GSR-consistent particles and constituent metals. This initial electrochemical screening has no effect on the integrity of the metallic particles, and SEM/EDX analysis conducted prior to and postvoltammetry show no differences in analytical output. The carbon tape is instrumental in retaining the GSR sample after electrochemical analysis, supported by comparison with orthogonal detection at a bare CSPE. This protocol shows great promise as a two-tier detection system for the presence of GSR from the hands of a subject, whereby initial screening can be conducted rapidly onsite by minimally trained operators; confirmation can follow at the same substrate to substantiate the voltammetric results.

  6. Examining assumptions regarding valid electronic monitoring of medication therapy: development of a validation framework and its application on a European sample of kidney transplant patients

    Directory of Open Access Journals (Sweden)

    Steiger Jürg

    2008-02-01

    Full Text Available Abstract Background Electronic monitoring (EM is used increasingly to measure medication non-adherence. Unbiased EM assessment requires fulfillment of assumptions. The purpose of this study was to determine assumptions needed for internal and external validity of EM measurement. To test internal validity, we examined if (1 EM equipment functioned correctly, (2 if all EM bottle openings corresponded to actual drug intake, and (3 if EM did not influence a patient's normal adherence behavior. To assess external validity, we examined if there were indications that using EM affected the sample representativeness. Methods We used data from the Supporting Medication Adherence in Renal Transplantation (SMART study, which included 250 adult renal transplant patients whose adherence to immunosuppressive drugs was measured during 3 months with the Medication Event Monitoring System (MEMS. Internal validity was determined by assessing the prevalence of nonfunctioning EM systems, the prevalence of patient-reported discrepancies between cap openings and actual intakes (using contemporaneous notes and interview at the end of the study, and by exploring whether adherence was initially uncharacteristically high and decreased over time (an indication of a possible EM intervention effect. Sample representativeness was examined by screening for differences between participants and non-participants or drop outs on non-adherence. Results Our analysis revealed that some assumptions were not fulfilled: 1 one cap malfunctioned (0.4%, 2 self-reported mismatches between bottle openings and actual drug intake occurred in 62% of the patients (n = 155, and 3 adherence decreased over the first 5 weeks of the monitoring, indicating that EM had a waning intervention effect. Conclusion The validity assumptions presented in this article should be checked in future studies using EM as a measure of medication non-adherence.

  7. Comparison of Lateral Flow Assay, Kidney Inhibition Swab, and Liquid Chromatography-Tandem Mass Spectrometry for the Detection of Penicillin G Residues in Sow Urine.

    Science.gov (United States)

    Shelver, Weilin L; Chakrabarty, Shubhashis; Smith, David J

    2017-03-01

    Sows (n = 126) were administered penicillin G; urine, collected at slaughter, was screened by kidney inhibition swab (KIS; 4 h testing time) and then stored at -80 °C (∼1200 days) until analysis by lateral flow assay (LF, ∼5 min testing time) and tandem quadrupole LC-MS/MS (TQ) analysis. The stability of penicillin in urine during storage was verified using TQ analyses. Quantitative results were well-correlated (R 2 = 0.98) with only a ∼10% decrease in penicillin concentration during the 3-year storage period. KIS retesting of stored samples returned results consistent with the original analyses. Lateral flow assay results were highly correlated with the KIS and TQ results. A KIS positive sample, which was not confirmed by TQ or LF, was assayed by Triple-TOF LC-MS to determine the cause of the apparent false positive. This study suggests LF can be used to quickly and efficiently screen for penicillin G residues before slaughter.

  8. Evaluation of the efficacy of real-time polymerase chain reaction for the routine early detection of Pseudomonas aeruginosa in cystic fibrosis sputum and throat swab specimens.

    LENUS (Irish Health Repository)

    Logan, Catriona

    2012-02-01

    A longitudinal study of 2099 sputa and throat swabs received from 183 pediatric cystic fibrosis patients over a 29-month period was used to evaluate the efficacy of real-time polymerase chain reaction (PCR) for the early detection of Pseudomonas aeruginosa as compared to microbiologic culture. Real-time PCR resulted in an increased number of specimens identified as P. aeruginosa positive. The sensitivity of culture was 82% (373\\/453) and of PCR was 93% (420\\/453) when considering both positive culture and PCR results as true positives. Of the 80 specimens identified as PCR positive\\/culture negative for P. aeruginosa, the subsequent patient sample in 32.5% (26\\/80) of specimens concerned was identified as P. aeruginosa culture positive, suggesting that PCR has the potential to detect P. aeruginosa earlier than the microbiologic culture. Real-time PCR analysis found no evidence of the Liverpool and Manchester epidemic P. aeruginosa strains in the cohort examined. The findings of this study highlight the importance of specimen collection protocols to ensure that adequate samples are received at the laboratory for testing, thereby minimizing the potential for reporting of false-negative P. aeruginosa culture results.

  9. The Comparison of Chlorhexidine Solution and Swab With Toothbrush and Toothpaste Effect on Preventing Oral Lesions in Hospitalized Patients in Intensive Care Unit

    National Research Council Canada - National Science Library

    Estaji, Zahra; Alinejad, Mohammad; Hassan Rakhshani, Mohammad; Rad, Mojtaba

    2016-01-01

    .... In this study, 30 patients were selected with target-based approach and equally divided into two groups through the permutation blocking method for oral care toothbrush and toothpaste and using chlorhexidine and swab...

  10. Improved diagnosis of Trichomonas vaginalis infection by PCR using vaginal swabs and urine specimens compared to diagnosis by wet mount microscopy, culture, and fluorescent staining

    NARCIS (Netherlands)

    C. van der Schee (Cindy); A.F. van Belkum (Alex); L. Zwijgers (Lisette); E. van der Brugge; E.L. O'Neill; A. Luijendijk (Ad); T. van Rijsoort-Vos; W.I. van der Meijden (Willem); J.F. Sluiters (Hans); H.A. Verbrugh (Henri)

    1999-01-01

    textabstractFour vaginal cotton swab specimens were obtained from each of 804 women visiting the outpatient sexually transmitted disease clinic of the Erasmus University Medical Center Rotterdam, Rotterdam, The Netherlands, for validation of various forms of Trichomonas

  11. Two, four, six, eight... stop and count before it is too late! An audit on swab, needle and instrument counts in theatre.

    LENUS (Irish Health Repository)

    Donnelly, T.

    2014-07-01

    A concurrent audit was conducted over a four week period to determine if the counting of swabs, needles and instruments for surgery adhered to local policy and recommended guidelines. Data were collected on 30 abdominal surgical procedures. This audit highlighted failings in the count process. It identified poor communication within the multidisciplinary team. There needs to be an increased awareness about local policy, national and international guidelines regarding the counting of swabs, needles and instruments for all surgical procedures.

  12. Comparison of 2 electronic cowside tests to detect subclinical ketosis in dairy cows and the influence of the temperature and type of blood sample on the test results.

    Science.gov (United States)

    Iwersen, M; Klein-Jöbstl, D; Pichler, M; Roland, L; Fidlschuster, B; Schwendenwein, I; Drillich, M

    2013-01-01

    The objective of this study was to determine the suitability of 2 electronic hand-held devices [FreeStyle Precision (FSP), Abbott GmbH & Co. KG, Wiesbaden, Germany and GlucoMen LX Plus (GLX), A. Menarini GmbH, Vienna, Austria] for measuring β-hydroxybutyrate (BHBA) in dairy cows. Three experiments were conducted to evaluate (1) the diagnostic performance of the devices, (2) the effect of the type of blood sample, and (3) the influence of the ambient temperature on the determined results. A total of 415 blood samples from lactating Holstein and Simmental cows were collected and analyzed with both devices (whole blood) and in a laboratory (serum). Correlation coefficients between whole-blood and serum BHBA concentrations were highly significant, with 94% for the FSP and 80% for the GLX device. Based on thresholds for subclinical ketosis of 1.2 and 1.4 mmol of BHBA/L, results obtained with the hand-held devices were evaluated by receiver operating characteristics analyses. This resulted in adjusted thresholds of 1.2 and 1.4 mmol/L for the FSP and 1.1 and 1.3 mmol/L for the GLX device. Applying these thresholds, sensitivities were 98 and 100% for the FSP and 80 and 86% for the GLX device, respectively. Corresponding specificities were 90 and 97% for the FSP and 87 and 96% for the GLX device, respectively. Additionally, concentrations of BHBA were tested with both devices in whole blood, EDTA-added whole blood, and in their resulting serum and plasma, collected from 65 animals. Determined BHBA concentrations were similar within each device for whole and EDTA-added blood, and in serum and plasma, but differed between whole blood and serum and between EDTA-added blood and plasma. Blood samples with low (0.4 mmol/L), medium (1.1 mmol/L), and high (1.6 mmol/L) BHBA concentrations were stored between +5 to +32°C and analyzed repeatedly at temperature levels differing by 4°C. Additionally, devices and test strips were stored at equal conditions and used for measurement

  13. A quantitative swab is a good non-invasive alternative to a quantitative biopsy for quantifying bacterial load in wounds healing by second intention in horses.

    Science.gov (United States)

    Van Hecke, L L; Hermans, K; Haspeslagh, M; Chiers, K; Pint, E; Boyen, F; Martens, A M

    2017-07-01

    The aim of this study was to evaluate different techniques for diagnosing wound infection in wounds healing by second intention in horses and to assess the effect of a vortex and sonication protocol on quantitative bacteriology in specimens with a histologically confirmed biofilm. In 50 wounds healing by second intention, a clinical assessment, a quantitative swab, a semi-quantitative swab, and a swab for cytology were compared to a quantitative tissue biopsy (reference standard). Part of the biopsy specimen was examined histologically for evidence of a biofilm. There was a significant, high correlation (P<0.001; r=0.747) between the outcome of the quantitative swabs and the quantitative biopsies. The semi-quantitative swabs showed a significant, moderate correlation with the quantitative biopsies (P<0.001; ρ=0.524). Higher white blood cell counts for cytology were significantly associated with lower log 10 colony-forming units (CFU) in the wounds (P=0.02). Wounds with black granulation tissue showed significantly higher log 10 CFU (P=0.003). Specimens with biofilms did not yield higher bacteriological counts after a vortex and sonication protocol was performed to release bacteria from the biofilm. Based on these findings, a quantitative swab is an acceptable non-invasive alternative to a quantitative biopsy for quantifying bacterial load in equine wounds healing by second intention. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Evaluation of the Essen Rotary as a new technique for bacterial swabs: results of a prospective controlled clinical investigation in 50 patients with chronic leg ulcers.

    Science.gov (United States)

    Al Ghazal, Philipp; Körber, Andreas; Klode, Joachim; Schmid, Ernst N; Buer, Jan; Dissemond, Joachim

    2014-02-01

    Most chronic wounds are colonised with different microorganisms, especially problematic bacteria like methicillin-resistant Staphylococcus aureus (MRSA), which represent an increasing therapeutic challenge in the modern wound therapy regimen. Therefore, it is essential to specify the bacteria in wounds for an individual-specific treatment. In most patients, an exemplary bacterial swab is taken from the centre of the wound surface. This so-called Levine technique is propagated currently as the gold standard. The aim of our clinical investigation was to compare the results of different swab techniques to the new established Essen Rotary. In this monocentric prospective investigation, 50 patients with chronic leg ulcers were examined consecutively. The results of our clinical study show that bacteria are heterogeneously spread on wound surfaces. The analysis of the semiquantitative measured results showed that the Essen Rotary could detect significant more bacteria with a total amount of 111 bacteria (P = 0·049) compared to usual swab techniques. Considerably, only the Essen Rotary identified five compared to three MRSA-patients detected by other techniques. The Essen Rotary is an efficient, economic and uncomplicated modification of bacteriological swab techniques which detects significant more bacteria compared to other conventional swab techniques. Therefore, the Essen Rotary may become the new gold standard in routinely taken bacteriological swabs especially for MRSA screenings in patients with chronic leg ulcers. © 2012 The Authors. International Wound Journal © 2012 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  15. Detection of group A streptococcal antigen directly from throat swabs with a ten-minute latex agglutination test.

    OpenAIRE

    Miceika, B G; Vitous, A S; Thompson, K D

    1985-01-01

    Results obtained with the Culturette brand 10-Minute Group A Strep ID system were compared with culture results to measure the ability of this system to detect group A streptococci directly from more than 800 throat swabs. Our study showed a sensitivity of 92.4% and a specificity of 92.8% for this acid extraction, latex agglutination method when compared with anaerobic culturing for group A streptococci. The results suggest that the 10-Minute Group A Strep ID method may prove to be a useful, ...

  16. Electronic Nicotine Delivery Systems (E-cigarette/Vape) use and Co-Occurring Health-Risk Behaviors Among an Ethnically Diverse Sample of Young Adults.

    Science.gov (United States)

    Lanza, H Isabella; Teeter, Heather

    2018-01-02

    Prevalence rates of electronic nicotine delivery systems (ENDS; i.e., e-cigarette/vape) use has grown exponentially in the past few years. College students present a particularly vulnerable group for ENDS use. The current study sought to expand the literature by examining the context in which college students use ENDS, co-occurring health risks beyond traditional tobacco use, and the role of ethnicity in ENDS use. Health-risk behavior survey data was collected from 452 undergraduates attending a large, public urban university during the 2015-2016 academic year. Ever ENDS users vs. non-ENDS users were compared across potential demographic, health-risk, and other health-related correlates. Almost 40% of participants reported lifetime use of ENDS. No ethnic or sex differences were found. The primary source for obtaining ENDS was friends and ENDS were most often used with friends vs. alone or with others not considered friends. Participants engaging in risky alcohol use and cigarette smoking had a higher likelihood of endorsing ENDS use. Conclusions/Importance: The current study indicated that a large proportion of college students have tried ENDS irrespective of ethnicity or sex. An increasingly normative social context may inform the popularity of ENDS use across ethnicity and sex, but additional research using ethnically diverse samples is warranted. Risky alcohol use appears to be a significant correlate of ENDS use, even after accounting for the robust relationship between ENDS use and cigarette smoking. The robust relationship between alcohol and tobacco use likely extends to ENDS use.

  17. Why You Shouldn't Use Cotton Swabs to Clean Your Ears

    Science.gov (United States)

    ... by using volume-limiting headphones when listening to music or watching videos on smartphones, tablets, and other electronic devices. Do the safety claims for these headphones hold up? Hunting and Other Shooting Sports Can Damage Your Hearing In rural areas, hearing ...

  18. The limitations of intimate samples in sexual offences.

    Science.gov (United States)

    Keating, S M

    1995-01-01

    This article reviews the problems of sampling and interpretation of scientific findings from samples taken by doctors during their examination of victims and suspects in sexual offences. The problems which may be encountered with vaginal, anal, oral and penile swabs are highlighted. These include contamination between samples and the consequences thereof. The responsibilities of the medical examiner, the investigating police officer and the scientist are stressed. Surveys of results stored in a database and case examples are used to illustrate the problems.

  19. Identification of Streptococcus parasanguinis DNA contamination in human buccal DNA samples.

    Science.gov (United States)

    Mahfuz, Istiak; Cheng, Wei; White, Stefan J

    2013-11-22

    The use of buccal swabs in clinical and scientific studies is a very popular method of collecting DNA, due to its non-invasive nature of collection. However, contamination of the DNA sample may interfere with analysis. Here we report the finding of Streptococcus parasanguinis bacterial DNA contamination in human buccal DNA samples, which led to preferential amplification of bacterial sequence with PCR primers designed against human sequence. Contamination of buccal-derived DNA with bacterial DNA can be significant, and may influence downstream genetic analysis. One needs to be aware of possible bacterial contamination when interpreting abnormal findings following PCR amplification of buccal swab DNA samples.

  20. Low-cost monitoring of Campylobacter in poultry houses by air sampling and quantitative PCR.

    Science.gov (United States)

    Søndergaard, M S R; Josefsen, M H; Löfström, C; Christensen, L S; Wieczorek, K; Osek, J; Hoorfar, J

    2014-02-01

    The present study describes the evaluation of a method for the quantification of Campylobacter by air sampling in poultry houses. Sampling was carried out in conventional chicken houses in Poland, in addition to a preliminary sampling in Denmark. Each measurement consisted of three air samples, two standard boot swab fecal samples, and one airborne particle count. Sampling was conducted over an 8-week period in three flocks, assessing the presence and levels of Campylobacter in boot swabs and air samples using quantitative real-time PCR. The detection limit for air sampling was approximately 100 Campylobacter cell equivalents (CCE)/m3. Airborne particle counts were used to analyze the size distribution of airborne particles (0.3 to 10 μm) in the chicken houses in relation to the level of airborne Campylobacter. No correlation was found. Using air sampling, Campylobacter was detected in the flocks right away, while boot swab samples were positive after 2 weeks. All samples collected were positive for Campylobacter from week 2 through the rest of the rearing period for both sampling techniques, although levels 1- to 2-log CCE higher were found with air sampling. At week 8, the levels were approximately 10(4) and 10(5) CCE per sample for boot swabs and air, respectively. In conclusion, using air samples combined with quantitative real-time PCR, Campylobacter contamination could be detected earlier than by boot swabs and was found to be a more convenient technique for monitoring and/or to obtain enumeration data useful for quantitative risk assessment of Campylobacter.

  1. The utility of polyester and cotton as swabbing substrates for the removal of cellular material from surfaces.

    Science.gov (United States)

    Mulligan, Christina M; Kaufman, Stacie R; Quarino, Lawrence

    2011-03-01

    Various types of cotton and polyester fabrics were tested to ascertain the optimal physical and chemical characteristics of fabrics needed for the removal of cellular material from surfaces. DNA quantitation values obtained on dried saliva stains showed no difference between cotton and polyester across all constructions and solvent conditions. Fabrics used dry and with water yielded higher quantitation values than those used with isopropanol. Quantitation values were also higher for wovens and nonwovens than knits across all solvent conditions. Low thread count fabrics used with water yielded higher quantitation values, but no correlation between thread count and quantitation values was observed with dry fabrics. A low thread count woven fabric, however, outperformed other tested fabrics when swabbing object surfaces in a highly used room. Full DNA profiles from fingerprints on glass surfaces were obtained with low thread count woven and nonwoven fabrics but not with the knit fabric tested. © 2011 American Academy of Forensic Sciences.

  2. Methicillin-resistant Staphylococcus Aureus Nasal Swab and Suction Drain Tip Cultures in 4573 Spinal Surgeries: Efficacy in Management of Surgical Site Infections.

    Science.gov (United States)

    Kawabata, Atsuyuki; Sakai, Kenichiro; Sato, Hirokazu; Sasaki, Shinichi; Torigoe, Ichiro; Tomori, Masaki; Yuasa, Masato; Matsukura, Yu; Arai, Yoshiyasu

    2017-08-01

    A retrospective single-center study. To assess the diagnostic value of methicillin-resistant Staphylococcus aureus (MRSA) nasal swab and suction drain tip cultures. The prognostic value of MRSA nasal swab and suction drain tip cultures has not been firmly established in spinal surgery. This study retrospectively included 4573 consecutive patients who underwent spinal surgery between January 2008 and December 2014. Patients diagnosed with infectious disease were excluded. Prophylactic antibiotics were administered intraoperatively and postoperatively for 48 h. MRSA nasal swab cultures were taken from all patients before surgery. Drains were removed when the volume of postoperative fluid drainage was less than 50 mL in the preceding 24 h and cultures were made. Surgical site infection (SSI) was defined according to Centers for Disease Control and Prevention criteria. SSI was identified in 94 cases (2.1%) and bacteria were isolated in 87 cases (92.6%). Positive MRSA nasal swab cultures were identified in 49 cases (1.1%). There was no significant difference in the SSI positivity rate between the MRSA nasal swab culture (+) and (-) groups. Positive drain tip cultures were found in 382 cases (8.4%), 28 of which developed SSI. There was a significant difference in the SSI positivity rate between the drain tip culture (+) and (-) groups. The sensitivity of drain tip culture was 29.8% and the specificity was 92.1%. In 16 of the 28 patients in the SSI (+) group with positive drain cultures, the same bacteria were isolated from the surgical site, giving a bacteria matching rate of 57.1%. MRSA nasal swab and drain tip cultures were not useful for predicting SSI. However, drain tip culture had a high positivity rate in the SSI group and the coincidence rate for the causative pathogen was relatively high. 4.

  3. Evaluation of a rapid method for the detection of streptococcal group A antigen directly from throat swabs.

    Science.gov (United States)

    Venezia, R A; Ryan, A; Alward, S; Kostun, W A

    1985-01-01

    Throat swabs from 196 pediatric patients were processed by a direct extraction-latex agglutination method (Group A Strep Direct Antigen Identification Test [DAI]) that detects group A streptococci in the specimen. The method requires a 45-min enzymatic extraction period at 37 degrees C and a 4-min reaction period with antibody-linked latex particles. The results were compared with those of the culture and fluorescent antibody methods and the clinical presentation of the patient for pharyngitis. Ninety-three percent of the specimens resulted in agreement by all tests, and 28% were culture positive for group A streptococci. Compared with the culture method, the DAI had a sensitivity and a specificity of 83% and 99%, respectively. The positive predictive values were 98% versus the culture method and 93% versus the fluorescent antibody method, whereas the negative predictive values were 94% versus both other methods. Of the 14 discrepant results when both clinical presentation of an acute pharyngitis and the test results were compared, the culture method provided the best correlation. An additional 64 specimens were processed by the DAI and another direct extraction-latex agglutination method (Culturette Ten-Minute Group A Strep ID Test), and the results were compared with those of the culture method. This group had a 40.6% culture isolation rate for group A streptococci. The sensitivity and specificity of the DAI and Strep ID methods versus the culture method were 81 and 100%, and 77 and 97%, respectively. These results indicate that the DAI is accurate for diagnosing group A streptococcal pharyngitis directly from throat swabs. However, negative results in the presence of a symptomatic patient must be confirmed by standard culture techniques. PMID:3884656

  4. Pressurised irrigation versus swabbing method in cleansing wounds healed by secondary intention: a randomised controlled trial with cost-effectiveness analysis.

    Science.gov (United States)

    Mak, Suzanne So-Shan; Lee, Man-Ying; Cheung, Jeanny Sui-Sum; Choi, Kai-Chow; Chung, Tak-Ki; Wong, Tze-Wing; Lam, Kit-Yee; Lee, Diana Tze-fan

    2015-01-01

    Wound cleansing should create an optimal healing environment by removing excess debris, exudates, foreign and necrotic material which are commonly present in the wounds that heal by secondary intention. At present, there is no research evidence for whether pressurised irrigation has better wound healing outcomes compared with conventional swabbing practice in cleansing wound. This study investigated the differences between pressurised irrigation and swabbing method in cleansing wounds that healed by secondary intention in relation to wound healing outcomes and cost-effectiveness. Multicentre, prospective, randomised controlled trial. The study took place in four General Outpatient Clinics in Hong Kong. Two hundred and fifty six patients with wounds healing by secondary intention were randomly assigned by having a staff independent of the study opening a serially numbered, opaque and sealed envelope to either pressurised irrigation (n=122) or swabbing (n=134). Staff undertaking study-related assessments was blinded to treatment assignment. Patients' wounds were followed up for 6 weeks or earlier if wounds had healed to determine wound healing, infection, symptoms, satisfaction, and cost effectiveness. The primary outcome was time-to-wound healing. Patients were analysed according to their treatment allocation. This trial is registered with ClinicalTrials.gov, number NCT01885273. Intention-to-treat analysis showed that pressurised irrigation group was associated with a shorter median time-to-wound healing than swabbing group [9.0 days (95% CI: 7.4-13.8) vs. 12.0 (95% CI: 10.2-13.8); p=0.007]. Pressurised irrigation group has significantly more patients experiencing lower grade of pain during wound cleansing (93.4% vs. 84.2%; p=0.02), and significantly higher median satisfaction with either comfort or cleansing method (MD 1 [95% CI: 5-6]; p=0.002; MD 1 [95% CI: 5-6]; pirrigation group and in 7 (5.2%) patients in swabbing group (p=0.44). Cost-effectiveness analysis

  5. Sponge and skin excision sampling for recovery of Salmonella and Campylobacter from defeathered broiler carcasses

    Science.gov (United States)

    Introduction: Salmonella and Campylobacter contamination of broiler carcass skin increases during feather removal. There are several methods for sampling carcasses including sponging or swabbing of skin surface and skin excision. It is unclear whether sponge sampling is adequate to remove bacteria f...

  6. Self-collected buccal swabs and rapid, real-time PCR during a large measles outbreak in Wales: Evidence for the protective effect of prior MMR immunisation.

    Science.gov (United States)

    Moore, Catherine; Cottrell, Simon; Hoffmann, Jörg; Carr, Michael; Evans, Hannah; Dunford, Linda; Lawson, Heather; Brown, Kevin E; Jones, Rachel

    2015-06-01

    We describe the laboratory response to a large measles outbreak that occurred during 2012-2013 centred in mid and west Wales, UK. To demonstrate the impact of rapid measles testing on the management of a large outbreak, to show the complex molecular epidemiology and determine the role of previous MMR immunisation on a large cohort of exposed people. Results from oral fluid antibody testing and self-collected buccal swabs tested by real-time PCR were reconciled and analysed to determine level of agreement and to calculate MMR vaccine efficacy during the outbreak. During the outbreak 1435 notifications of measles were received from across Wales. Samples were received from 70% of notified cases with a positivity rate of 56% within the outbreak compared to 15% for the rest of Wales. Measles RNA was detected in 53 cases with previous history of MMR immunisation, but viral loads were lower than those detected in unimmunised cases. The molecular epidemiology showed at least two distinct D8 strains of measles virus were introduced into Wales along with a separate introduction of a B3 strain outside the outbreak area. Molecular testing of all notified measles cases offers the most rapid way of confirming the introduction of measles into a population potentially before secondary transmission has already occurred. The outbreak data confirms the protective effect of the MMR vaccine with vaccine efficacy calculated at 96% for one dose and 99% for two doses supporting the WHO recommendations for a two dose MMR immunisation schedule. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Evaluation of GeneXpert® system for detection of methicillin-resistant Staphyloccocus aureus in clinical samples

    Directory of Open Access Journals (Sweden)

    Antonella Mencacci

    2011-03-01

    Full Text Available Infections caused by methicillin-resistant Staphyloccocus aureus strains (MRSA have reached epidemic proportions globally, being the major cause of nosocomial infections. Rapid identification of MRSA in nasal swabs or in clinical samples is considered a useful strategy for control and treatment of these infections. GeneXpert system (Cepheid Europe,Vira-Solelch, Maurence-Scopont-France can detect by real-time PCR in approximately one hour methicillin-resistant S. aureus or coagulase-negative staphylococci (CoNS in clinical samples, in comparison with 24 hours for the culture or 48 hours for the antimicrobial susceptibility testing. In this study GeneXpert system was compared with traditional tests for MRSA detection in nasal swabs, bloodcultures and surgical wound swabs. Materials and methods. Eighteen nasal swabs, 23 blood-cultures and 13 surgical wound swabs were tested. The samples were cultured on blood-agar and mannitol-salt agar. Identification of isolates was carried out with traditional tests (Gram staining, catalase, coagulase and automatic Phoenix system. Methicillin-susceptibility was evaluated according to 2010 CLSI guidelines. GeneXpert system was performed according to manufacturers instructions, by using the specific kits and methicillin-resistance was detected by amplification of the genic sequences spa, SCC e mecA. Results. The results showed a 100% accordance between GeneXpert system and traditional tests for detection of methicillin-resistant staphylococci. In particular, among 18 nasal swabs, no MRSA was detected, while 1 bloodculture (4.3% and 4 surgical wound swabs (30.7% were positive for MRSA. Conclusions. GeneXpert system allows a rapid detection of MRSA in clinical samples and shows the same sensitivity and specificity as traditional tests. Therefore, it represents a further effective diagnostic method for prevention and treatment of nosocomial infections due to methicillin-resistant staphylococci.

  8. Simultaneous analysis of polychlorinated biphenyls and organochlorine pesticides in seawater samples by membrane-assisted solvent extraction combined with gas chromatography-electron capture detector and gas chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Shi, Xizhi; Tang, Zigang; Sun, Aili; Zhou, Lei; Zhao, Jian; Li, Dexiang; Chen, Jiong; Pan, Daodong

    2014-12-01

    A highly efficient and environment-friendly membrane-assisted solvent extraction system combined with gas chromatography-electron capture detector was applied in the simultaneous determination of 17 polychlorinated biphenyls and organochlorine pesticides in seawater samples. Variables affecting extraction efficiency, including extraction solvent used, stirring rate, extraction time, and temperature, were optimized extensively. Under optimal extraction conditions, recoveries between 76.9% and 104.6% in seawater samples were achieved, and relative standard deviation values below 10% were obtained. The limit of detection (signal-to-noise ratio=3) and limit of quantification (signal-to-noise ratio=10) of 17 polychlorinated biphenyls and organochlorine pesticides in seawater ranged from 0.14ngL(-1) to 0.36ngL(-1) and 0.46ngL(-1) to 1.19ngL(-1), respectively. Matrix effects on extraction efficiency were evaluated by comparing with the results obtained using tap water. The extraction effect of developed membrane-assisted solvent extraction method was further demonstrated by gas chromatography-tandem mass spectrometry which can provide structural information of the analytes for more accurate identification, and results identical to those produced by gas chromatography-electron capture detector were obtained. These findings demonstrate the applicability of the developed membrane-assisted solvent extraction determination method for coupling to gas chromatography-electron capture detector or tandem mass spectrometry for determining polychlorinated biphenyls and organochlorine pesticides in seawater samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Relative quantification and detection of different types of infectious bursal disease virus in bursa of Fabricius and cloacal swabs using real time RT-PCR SYBR green technology

    DEFF Research Database (Denmark)

    Li, Yiping; Handberg, K.J.; Kabell, Susanne

    2007-01-01

    In present study, different types of infectious bursal disease virus (IBDV), virulent strain DK01, classic strain F52/70 and vaccine strain D78 were quantified and detected in infected bursa of Fabricius (BF) and cloacal swabs using quantitative real time RT-PCR with SYBR green dye. For selection...

  10. Application of kidney inhibition swab tests to evaluate penicillin-G residues in sow tissues and body fluids following intramuscular injection

    Science.gov (United States)

    Kidney inhibition swab (KIS) tests, recently adapted by the US FSIS for antibiotics on-site screening, were employed to evaluate the depletion of penicillin-G residues from kidney, liver, muscle, serum, and urine of sows after intramuscular (IM) penicillin-G procaine administration. Sows (n=130; 22...

  11. Development of a real time PCR for the detection of Taylorella equigenitalis directly from genital swabs and discrimination from Taylorella asinigenitalis

    NARCIS (Netherlands)

    Wakeley, P.R.; Errington, J.; Hannon, S.; Roest, H.I.J.; Carson, T.; Hunt, B.; Sawyer, J.; Heath, P.

    2006-01-01

    discriminatory real time PCR for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM), and the related species T. asinigenitalis was developed for the direct examination of genital swabs. The 112 bp amplicons produced from the two species were

  12. Clinical utility of a nasal swab methicillin-resistant Staphylococcus aureus polymerase chain reaction test in intensive and intermediate care unit patients with pneumonia.

    Science.gov (United States)

    Giancola, Stephanie E; Nguyen, Ai Thi; Le, Binh; Ahmed, Omar; Higgins, Catherine; Sizemore, James A; Orwig, Kara W

    2016-11-01

    This retrospective study aimed to validate the concordance between nasal swab methicillin-resistant Staphylococcus aureus (MRSA) polymerase chain reaction (PCR) test and respiratory culture and to determine the number of potentially preventable days of anti-MRSA therapy in patients with pneumonia. Two hundred adult inpatients in the intensive and intermediate care units were included. The nasal swab MRSA PCR test was positive in 55 (27.5%) patients. MRSA was isolated from respiratory culture in 21 (10.5%) patients. The nasal swab MRSA PCR test demonstrated 90.5% sensitivity, 79.9% specificity, 34.5% positive predictive value, and 98.6% negative predictive value. Anti-MRSA therapy was initiated in 168 (84%) patients. Patients in the study received a combined 782days of anti-MRSA therapy; 300days were considered potentially preventable. This study suggests that the nasal swab MRSA PCR test may be used to guide discontinuation of anti-MRSA antibiotics in patients with clinically confirmed pneumonia in the intensive or intermediate care units. Published by Elsevier Inc.

  13. Immunoexpression of cytokeratin 19 in oral swab from fixed orthodontic appliance users

    Directory of Open Access Journals (Sweden)

    Dwi Mutia Ramdhini

    2007-11-01

    Full Text Available The use of fixed orthodontic appliances can improve someone's mastication, speech and appearance. However, this appliance acts as a strange object that may cause irritation to the mucosa epithelial of oral cavity, because of the friction and pressure from the components of the fixed orthodontic appliances which are in direct contact with the oral mucosa. Irritation in the oral mucosa could stimulate the increase of cytokeratin. The appearance of cytokeratin is then used to identify the condition of these cells. This study was a descriptive research to find the expression of cytokeratin 19 with immunohistochemical method in oral mucosa epithelial of fixed orthodontic appliances users. Sample in this study was chosen from 30 fixed orthodontic appliances users. The result of this study was determined by calculating the number of positive cells (brown, compared with total number of cells. The account of positive cells would present the reaction of the epithelial cells according to the inflamation stage which caused by the use of orthodontic appliances. As a conclusion of this study, the use of fixed orthodontic appliances may cause changes in epithelial mucosa which form an adaptation process by increasing the number of progenitor cells marked by cytokeratin 19.

  14. Being an Informed Consumer of Health Information and Assessment of Electronic Health Literacy in a National Sample of Internet Users: Validity and Reliability of the e-HLS Instrument

    Science.gov (United States)

    Yeatts, Dale; Hughes, Susan; Hudson, Cassie; Bell, Valarie

    2016-01-01

    Background The Internet, with its capacity to provide information that transcends time and space barriers, continues to transform how people find and apply information to their own lives. With the current explosion in electronic sources of health information, including thousands of websites and hundreds of mobile phone health apps, electronic health literacy is gaining an increasing prominence in health and medical research. An important dimension of electronic health literacy is the ability to appraise the quality of information that will facilitate everyday health care decisions. Health information seekers explore their care options by gathering information from health websites, blogs, Web-based forums, social networking websites, and advertisements, despite the fact that information quality on the Internet varies greatly. Nonetheless, research has lagged behind in establishing multidimensional instruments, in part due to the evolving construct of health literacy itself. Objective The purpose of this study was to examine psychometric properties of a new electronic health literacy (ehealth literacy) measure in a national sample of Internet users with specific attention to older users. Our paper is motivated by the fact that ehealth literacy is an underinvestigated area of inquiry. Methods Our sample was drawn from a panel of more than 55,000 participants maintained by Knowledge Networks, the largest national probability-based research panel for Web-based surveys. We examined the factor structure of a 19-item electronic Health Literacy Scale (e-HLS) through exploratory factor analysis (EFA) and confirmatory factor analysis, internal consistency reliability, and construct validity on sample of adults (n=710) and a subsample of older adults (n=194). The AMOS graphics program 21.0 was used to construct a measurement model, linking latent factors obtained from EFA with 19 indicators to determine whether this factor structure achieved a good fit with our entire sample

  15. A modified ‘NanoSuit®’ preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy

    Science.gov (United States)

    Takaku, Yasuharu; Suzuki, Hiroshi; Kawasaki, Hideya; Ohta, Isao; Ishii, Daisuke; Hirakawa, Satoshi; Tsutsui, Takami; Matsumoto, Haruko; Takehara, Sayuri; Nakane, Chinatsu; Sakaida, Kana; Suzuki, Chiaki; Muranaka, Yoshinori; Kikuchi, Hirotoshi; Konno, Hiroyuki; Shimomura, Masatsugu

    2017-01-01

    Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10−3 to 10−7 Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the ‘NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved ‘NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the ‘real' organization of cells and their functions. PMID:28405375

  16. A modified 'NanoSuit®' preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy.

    Science.gov (United States)

    Takaku, Yasuharu; Suzuki, Hiroshi; Kawasaki, Hideya; Ohta, Isao; Ishii, Daisuke; Hirakawa, Satoshi; Tsutsui, Takami; Matsumoto, Haruko; Takehara, Sayuri; Nakane, Chinatsu; Sakaida, Kana; Suzuki, Chiaki; Muranaka, Yoshinori; Kikuchi, Hirotoshi; Konno, Hiroyuki; Shimomura, Masatsugu; Hariyama, Takahiko

    2017-03-01

    Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10-3 to 10-7 Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the 'NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved 'NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the 'real' organization of cells and their functions.

  17. Simulation of surge and swab pressures in well drilling operations; Simulacao do problema de 'surge' e 'swab' em atividades de perfuracao de pocos de petroleo

    Energy Technology Data Exchange (ETDEWEB)

    Almeida, Fernando T.G.M.C. de; Kimura, Hudson Faglioni; Ramalho, Vanessa; Negrao, Cezar O. Ribeiro; Junqueira, Silvio L.M. [Universidade Tecnologica Federal do Parana (UTFPR), Curitiba, PR (Brazil); Martins, Andre Leibsohn [PETROBRAS, Rio de Janeiro, RJ (Brazil)

    2008-07-01

    The movements of the well drilling pipe, known as trips, cause variations in the well inner pressure. When the pipe is moving downwards, in an operation called 'running', the pressure increases and is known as surge pressure and, when it is moving upwards, in an operation called 'pulling', the pressure decreases and it is so called a swab pressure. The study of this phenomenon is of great importance not only due to financial reasons but also for the determination of speeds and accelerations which should be used in running and pulling operations. Among the researchers have studied this problem, Fontenot and Clark are two of the most important. They formulated the problem solution through considerations about the friction factor. The present work's target is to develop a computational program which allows the calculus of those pressures, according to previous investigations and models found in the literature and for different types of fluids as well, such as Bingham fluid and Power Law fluid. (author)

  18. Electronics and electronic systems

    CERN Document Server

    Olsen, George H

    1987-01-01

    Electronics and Electronic Systems explores the significant developments in the field of electronics and electronic devices. This book is organized into three parts encompassing 11 chapters that discuss the fundamental circuit theory and the principles of analog and digital electronics. This book deals first with the passive components of electronic systems, such as resistors, capacitors, and inductors. These topics are followed by a discussion on the analysis of electronic circuits, which involves three ways, namely, the actual circuit, graphical techniques, and rule of thumb. The remaining p

  19. Temperature distribution in a sample with second-phase microinclusions during irradiation by a low-energy high-current pulsed electron beam

    Science.gov (United States)

    Shepel', D. A.; Markov, A. B.

    2017-02-01

    Using the methods of numerical integration, a temperature field has been calculated that arose in the surface layer of titanium nickelide target with NiTi2 intermetallic inclusions during irradiation by a lowenergy high-current electron beam with a duration of the order of a microsecond. The calculated temperature field has been compared with that obtained previously for a target of stainless steel 316L containing MnS inclusions. It has been found that, as in the case of stainless steel, the regions of inclusions are overheated. However, the temperature increase for NiTi2 (12 K) is significantly lower than in the case of stainless steel 316L (283 K). The dynamics of melting of these systems are also considerably different.

  20. Noninvasive stratum corneum sampling and electron microscopical examination of skin barrier integrity: pilot study with a topical glycerin formulation for atopic dermatitis.

    Science.gov (United States)

    Daehnhardt-Pfeiffer, S; Surber, C; Wilhelm, K-P; Daehnhardt, D; Springmann, G; Boettcher, M; Foelster-Holst, R

    2012-01-01

    Therapy of atopic dermatitis encloses use of medicated and nonmedicated preparations. Results of clinical and biophysical procedures indirectly describe the condition of the impaired skin barrier (SB). Direct evaluation of SB integrity is only possible by electron microscopical visualization, e.g. intercellular lipid lamellae (ICLL) organization of the stratum corneum. SB integrity was measured by morphometric analysis of ICLL in healthy and atopic skin and after a 15-day treatment (plus 7-day follow-up) of atopic skin with a glycerin preparation. Significant treatment effect was shown by the restoration of the ICLL. The study reveals that morphometric analysis of ICLL organization is suitable to differentiate between healthy and diseased skin and to semiquantitatively determine the effect of a nonmedicated glycerin formulation. Small treatment cohort. Copyright © 2012 S. Karger AG, Basel.

  1. Diagnostic herd sensitivity using environmental samples

    DEFF Research Database (Denmark)

    Vigre, Håkan; Josefsen, Mathilde Hartmann; Seyfarth, Anne Mette

    . In our example, the prevalence of infected pigs in each herd was estimated from the pooled samples of nasal swabs. Logistic regression was used to estimate the effect of animal prevalence on the probability to detect MRSA in the dust and air samples at herd level. The results show a significant increase...... of the within herd prevalence, and performed almost perfectly at a prevalence of 25% infected pigs (sensitivity=99%). In general, the dependence of within herd prevalence should be considered in designing surveillance programs based on environmental samples.......Due to logistic and economic benefits, the animal industry has an increased interest in using environmental samples to classify herds free of infections. For a valid interpretation of results obtained from environmental samples, the performance of the diagnostic method using these samples must...

  2. A novel graphene nanosheets coated stainless steel fiber for microwave assisted headspace solid phase microextraction of organochlorine pesticides in aqueous samples followed by gas chromatography with electron capture detection.

    Science.gov (United States)

    Ponnusamy, Vinoth Kumar; Jen, Jen-Fon

    2011-09-28

    In this study, a novel graphene nanosheets (GNSs) coated solid phase microextraction (SPME) fiber was prepared by immobilizing microwave synthesized GNSs on a stainless steel wire. Microwave synthesized GNSs were verified by X-ray diffraction, field emission-scanning electron microscopy (FE-SEM) and transmission electron microscope (TEM). GNS-SPME fiber was characterized using FE-SEM and the results showed the GNS coating was homogeneous, porous, and highly adherent to the surface of the stainless steel fiber. The performance and feasibility of the GNS-SPME fiber was evaluated under one-step microwave assisted (MA) headspace (HS) SPME followed by gas chromatography with electron capture detection for five organochlorine pesticides (OCPs) in aqueous samples. Parameters influencing the extraction efficiency of MA-HS-GNS-SPME such as microwave irradiation power and time, pH, ionic strength, and desorption conditions were thoroughly examined. Under the optimized conditions, detection limits for the OCPs varied between 0.16 and 0.93 ng L(-1) and linear ranges varied between 1 and 1500 n gL(-1), with correlation coefficients ranging from 0.9984 to 0.9998, and RSDs in the range of 3.6-15.8% (n=5). In comparison with the commercial 100 μm polydimethylsiloxane fiber, the GNS coated fiber showed better extraction efficiency, higher mechanical and thermal stability (up to 290°C), longer life span (over 250 times), and lower production cost. The method was successfully applied to the analysis of real water samples with recoveries ranged between 80.1 and 101.1% for river water samples. The results demonstrated that the developed MA-HS-GNS-SPME method was a simple, rapid, efficient pretreatment and environmentally friendly procedure for the analysis of OCPs in aqueous samples. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Immunodetection of rasP21 and c-myc oncogenes in oral mucosal swab preparation from clove cigarette smokers

    Directory of Open Access Journals (Sweden)

    Silvi Kintawati

    2008-12-01

    Full Text Available Background: Smoking is the biggest factor for oral cavity malignancy. Some carcinogens found in cigar will stimulate epithel cell in oral cavity and cause mechanism disturbance on tissue resistance and produce abnormal genes (oncogenes. Oncogenes ras and myc are found on malignant tumor in oral cavity which are associated with smoking. Purpose: This research is to find the expression of oncogenes rasP21 and c-myc in oral mucosa epithelial of smoker with immunocytochemistry reaction. Methods: An oral mucosal swab was performed to 30 smokers categorized as light, moderate, and chain, and 10 non smokers which was followed by immunocytochemistry reaction using antibody towards oncogene rasP21 and c-myc is reacted to identify the influence of smoking towards malignant tumor in oral cavity. The result is statistically analyzed using Kruskal-Wallis test. Result: Based on the observation result of oncogene rasP21reaction, it shows that there is significant difference between non smoker group and light smoker, compared to moderate and chain smoker group (p < 0.01. On the other side, the observation result of oncogene c-myc indicates that there is no significant difference between the group of non smokers and the group of light, moderate, and chain smokers (p > 0.05. Conclusion: The higher the possibility of oral cavity malignancy and that the antibody for rasP21 oncogene can be used as a marker for early detection of oral cavity malignancy caused by smoking.

  4. Some virulence genes of Escherichia coli isolated from cloacal swabs of healthy Alagoas Curassows (Pauxi mitu in Brazil

    Directory of Open Access Journals (Sweden)

    André A.B. Saidenberg

    2013-04-01

    Full Text Available Birds of the Cracidae family (curassows, guans, and chachalacas are endemic of the Neotropics and 50 species are currently classified. Brazil has 22 species, seven of which are considered threatened. The Alagoas Curassow (Pauxi mitu species is considered extinct in the wild; but about 120 birds are alive in captivity. Conservation of this species depends entirely on correct management. Health reports of both wildlife and captive curassows are rare. In this study the presence of Escherichia coli was evaluated in 23 healthy Alagoas Curassows from two private breeding centres. E. coli was isolated from cloacal swabs, and the presence of genes encoding cytotoxic necrotising factor 1 (cnf1, alpha-haemolysin (hly, aerobactin (iuc, serum resistance (iss and the following adhesions: S fimbriae (sfa, pili associated with pyelonephritis (pap and temperature-sensitive haemagglutinin (tsh were investigated. E. coli was isolated from 78.3% (18/23 of the birds, and the percentage of curassows colonized by E. coli was similar between the two facilities. From the 22 E. coli isolates, 15 (68.2% were positive for at least one virulence factor by PCR, and the most frequently found gene was iss (50%. No curassows had clinical signs of disease. Nevertheless, the presence of some E. coli strains may be a concern to the wildlife in captivity. Additional health surveillance studies are essential to guarantee successful conservation programmes for threatened cracids in Brazil.

  5. Healthy Farms in Slovakia: Antibiotic Sensitivity of Escherichia coli Isolated from Rectal Swabs of Chicken and Ram

    Directory of Open Access Journals (Sweden)

    Jaroslav Pochop

    2012-10-01

    Full Text Available The aim of this experiment was determine of antibiotic resistance profile of Escherichia coli isolated from rectal swabs of chicken and ram from two different conventional breeding from Slovakia. For the antibiotic susceptibility testing disk diffusion method was used. A tested bacterium, Escherichia coli was exposed against four antibiotics: amikacin, gentamycin, tetracycline and chloramphenicol. For the identification of this strain, we used Chromogenic coliform agar, Triple sugar iron agar and biochemical test (ENTEROtest 24. For genetic identification of Escherichia coli Step One Real Time PCR with using special primer was used. Was determined that antibiotic resistance in Escherichia coli was not found. Was found susceptibility in all cases of Escherichia coli isolates. Antibiotic resistance is a biological danger. Bacteria, which we study, are considered to reservoirs of resistant genes and they are facultative and obligate pathogens. If these pathogen bacteria cause diseases those these diseases are difficult to treat. In this study, we determined that we have healthy farms in Slovakia too. In this farm antibiotic was not use and we do not determined any resistance to antibiotics, which we used in experiment.

  6. How is the high vaginal swab used to investigate vaginal discharge in primary care and how do GPs' expectations of the test match the tests performed by their microbiology services?

    Science.gov (United States)

    Noble, H; Estcourt, C; Ison, C; Goold, P; Tite, L; Carter, Y H

    2004-06-01

    To describe the management of vaginal discharge in general practice, with particular regard to the use of the high vaginal swab (HVS), and to compare GPs' expectations of this test with the processing and reporting undertaken by different laboratories. A postal questionnaire survey of 2146 GPs in the North Thames area and postal questionnaire study of the 22 laboratories serving the same GPs were carried out. GPs were asked how they would manage a young woman with vaginal discharge and what information they would like on an HVS report. Laboratories were asked how they would process and report on the HVS sample from the same patient. Response rate was 26%. 72% of GPs would take an HVS and 62% would refer on to a genitourinary medicine (GUM) clinic. 45% would offer empirical therapy and 47% of these would treat for candida initially. 75% of GPs routinely request "M,C&S" on HVS samples but 55% only want to be informed about specific pathogens. Routine processing of HVS samples varies widely between laboratories and 86% only report specific pathogens. 78% of GPs would like to be offered a suggested diagnosis on HVS reports, and 74% would like a suggested treatment. 43% of laboratories ever provide a diagnosis, and 14% provide a suggested treatment. GPs frequently manage vaginal discharge and most of them utilise the HVS. GPs' expectations of the test are not well matched to laboratory processing or reporting of the samples.

  7. The comparative utility of oral swabs and probang samples for detection of foot-and-mouth disease virus infection in cattle and pigs

    DEFF Research Database (Denmark)

    Stenfeldt, Anna Carolina; Lohse, Louise; Belsham, Graham

    2013-01-01

    Foot-and-mouth disease virus (FMDV) RNA was measured using quantitative reverse transcription-PCR (qRT-PCR) assays in oralswab and probangsamples collected from cattle and pigs during experimental infections with serotype O FMDV. During acute infection, FMDV RNA was measurable in oralswabs as wel...

  8. Feeding of waste milk to Holstein calves affects antimicrobial resistance of Escherichia coli and Pasteurella multocida isolated from fecal and nasal swabs.

    Science.gov (United States)

    Maynou, G; Bach, A; Terré, M

    2017-04-01

    The use of milk containing antimicrobial residues in calf feeding programs has been shown to select for resistant fecal Escherichia coli in dairy calves. However, information is scarce about the effects of feeding calves waste milk (WM) on the prevalence of multidrug-resistant bacteria. The objective of this study was to determine the antimicrobial resistance patterns of fecal E. coli and nasal Pasteurella multocida isolates from calves fed either milk replacer (MR) or WM in 8 commercial dairy farms (4 farms per feeding program). Fecal and nasal swabs were collected from 20 ± 5 dairy calves at 42 ± 3.2 d of age, and from 10 of these at approximately 1 yr of age in each study farm to isolate the targeted bacteria. Furthermore, resistance of E. coli isolates from calf-environment and from 5 calves at birth and their dams was also evaluated in each study farm. Resistances were tested against the following antimicrobial agents: amoxicillin-clavulanic acid, ceftiofur, colistin, doxycycline (DO), enrofloxacin (ENR), erythromycin, florfenicol, imipenem, and streptomycin. A greater number of fecal E. coli resistant to ENR, florfenicol, and streptomycin and more multidrug-resistant E. coli phenotypes were isolated in feces of calves fed WM than in those fed MR. However, the prevalence of fecal-resistant E. coli was also influenced by calf age, as it increased from birth to 6 wk of age for ENR and DO and decreased from 6 wk to 1 yr of age for DO regardless of the feeding program. From nasal samples, an increase in the prevalence of colistin-resistant P. multocida was observed in calves fed WM compared with those fed MR. The resistance patterns of E. coli isolates from calves and their dams tended to differ, whereas similar resistance profiles among E. coli isolates from farm environment and calves were observed. The findings of this study suggest that feeding calves WM fosters the presence of resistant bacteria in the lower gut and respiratory tracts of dairy calves

  9. The effect of sampling and storage on the fecal microbiota composition in healthy and diseased subjects.

    Directory of Open Access Journals (Sweden)

    Danyta I Tedjo

    Full Text Available Large-scale cohort studies are currently being designed to investigate the human microbiome in health and disease. Adequate sampling strategies are required to limit bias due to shifts in microbial communities during sampling and storage. Therefore, we examined the impact of different sampling and storage conditions on the stability of fecal microbial communities in healthy and diseased subjects. Fecal samples from 10 healthy controls, 10 irritable bowel syndrome and 8 inflammatory bowel disease patients were collected on site, aliquoted immediately after defecation and stored at -80 °C, -20 °C for 1 week, at +4°C or room temperature for 24 hours. Fecal transport swabs (FecalSwab, Copan were collected and stored for 48-72 hours at room temperature. We used pyrosequencing of the 16S gene to investigate the stability of microbial communities. Alpha diversity did not differ between all storage methods and -80 °C, except for the fecal swabs. UPGMA clustering and principal coordinate analysis showed significant clustering by test subject (p < 0.001 but not by storage method. Bray-Curtis dissimilarity and (unweighted UniFrac showed a significant higher distance between fecal swabs and -80 °C versus the other methods and -80 °C samples (p < 0.009. The relative abundance of Ruminococcus and Enterobacteriaceae did not differ between the storage methods versus -80 °C, but was higher in fecal swabs (p < 0.05. Storage up to 24 hours (at +4 °C or room temperature or freezing at -20 °C did not significantly alter the fecal microbial community structure compared to direct freezing of samples from healthy subjects and patients with gastrointestinal disorders.

  10. Rapid Diagnosis of Trichomonas vaginalis by Testing Vaginal Swabs in an Isothermal Helicase-Dependent AmpliVue Assay.

    Science.gov (United States)

    Gaydos, Charlotte A; Hobbs, Marcia; Marrazzo, Jeanne; Schwebke, Jane; Coleman, Jenell S; Masek, Billie; Dize, Laura; Jang, Dan; Li, Jenny; Chernesky, Max

    2016-06-01

    The AmpliVue Trichomonas Assay (Quidel) is a new Federal Drug Administration-cleared rapid test for qualitative detection of Trichomonas vaginalis (TV) DNA in female vaginal specimens. The assay is based on BioHelix's helicase-dependent amplification isothermal technology in conjunction with a disposable lateral-flow detection device, with a total turnaround time of approximately 45 minutes. The objective of this study was to compare the performance of this new assay to wet preparation and culture as well as to another Federal Drug Administration-cleared nucleic acid amplification assay. Four clinician collected vaginal swabs were obtained from women attending sexually transmitted disease, family planning, and OB/GYN clinics and tested by AmpliVue Trichomonas Assay and comparator tests: saline microscopy, TV culture (InPouch), and Aptima TV. AmpliVue Trichomonas Assay results were compared with a composite positive comparator (CPC) as determined by the results from culture and/or wet mount microscopic examination. At least one of either the wet preparation or culture reference test results was required to be positive to establish CPC. A total of 992 patients, 342 symptomatic and 650 asymptomatic patients, were included in the study. Results for AmpliVue for all women combined compared with saline microscopy and culture as a CPC yielded a sensitivity of 100%. Specificity for all women was 98.2%. Overall percent agreement versus Aptima TV was 97.8%. Sensitivity for AmpliVue compared with Aptima was 90.7% %, whereas specificity was 98.9%. The rapid AmpliVue Trichomonas Assay performed as well as microscopy and culture, and had comparable sensitivity and specificity to another nucleic acid amplification test for the detection of TV. This study provided evidence of new diagnostic options and indicated very good performance of amplified testing for detection of TV in symptomatic and asymptomatic women.

  11. Rapid Diagnosis of Trichomonas vaginalis by Testing Vaginal Swabs in an Isothermal Helicase-Dependent AmpliVue™ Assay

    Science.gov (United States)

    Gaydos, Charlotte A.; Hobbs, Marcia; Marrazzo, Jeanne; Schwebke, Jane; Coleman, Jenell S.; Masek, Billie; Dize, Laura; Jang, Dan; Li, Jenny; Chernesky, Max

    2016-01-01

    Background The AmpliVue™ Trichomonas Assay (Quidel) is a new FDA cleared rapid test for qualitative detection of Trichomonas vaginalis (TV) DNA in female vaginal specimens. The assay is based on BioHelix’s Helicase-Dependent Amplification (HDA) isothermal technology in conjunction with a disposable lateral-flow detection device, with a total turn-around time of approximately 45 minutes. Objective The objective of this study was to compare the performance of this new assay to wet preparation and culture, as well as to another FDA cleared nucleic acid amplification assay. Methods Four clinician collected vaginal swabs were obtained from women attending STD, family planning, and OB/GYN clinics and tested by AmpliVue™ Trichomonas Assay and comparator tests: saline microscopy, TV culture (InPouch™), and Aptima® TV (ATV). AmpliVue™ Trichomonas Assay results were compared to a composite positive comparator (CPC) as determined by the results from culture and/or wet mount microscopic examination. At least one of either the wet preparation or culture reference test results was required to be positive to establish CPC. Results A total of 992 patients, 342 symptomatic and 650 asymptomatic patients, were included in the study. Results for AmpliVue for all women combined compared to saline microscopy and culture as a composite positive comparator yielded a sensitivity of 100%. Specificity for all women was 98.2%. Overall percent agreement versus Aptima® TV was 97.8%. Sensitivity for AmpliVue compared to Aptima® was 90.7% %, while specificity was 98.9%. Conclusions The rapid AmpliVue™ Trichomonas Assay performed as well as microscopy and culture, and had comparable sensitivity and specificity to another NAAT for the detection of TV. This study provided evidence of new diagnostic options and indicated very good performance of amplified testing for detection of TV in symptomatic and asymptomatic women. PMID:27196258

  12. Resistance to Antibiotics in Strains of Staphylococcus spp., Enterococcus spp. and Escherichia coli Isolated from Rectal Swabs of Pigs

    Directory of Open Access Journals (Sweden)

    M. Kolář

    2008-01-01

    Full Text Available The study aimed at determining the level of resistance of selected bacterial species (Staphylococcus spp., Enterococcus spp., Escherichia coli isolated from rectal swabs of pigs to antimicrobial agents. The tested strains were isolated from piglets aged 7 to 30 days. Bacterial species were identified by standard microbiological techniques and susceptibility to antibiotics was determined quantitatively by the standard microdilution method. Resistance of the Staphylococcus aureus strain to oxacillin was confirmed by detection of the mecA gene and PBP2a. A total of 115 Staphylococcus spp. isolates were collected. In the case of Staphylococcus aureus, the methicillin-resistant strain (MRSA was identified. Moreover, higher frequency of coagulase-negative staphylococci with minimum inhibitory concentration of oxacillin ≥ 0.5 mg/l was noticed. Inducible resistance to clindamycin in the Staphylococcus hominis strain was also detected. The strains of Enterococcus spp. (61 isolates exhibited high resistance to tetracycline (98.5%, erythromycin (86.8% and chloramphenicol (54.4%. Vancomycin-resistant enterococci were not isolated. In the case of Escherichia coli strains (111 isolates, higher frequency of resistant strains to tetracycline (81.1% and ampicillin (62.2% was documented. Resistance to fluoroquinolones and production of broad-spectrum β-lactamases was not noticed. The presented study may be considered as a pilot project assessing the prevalence of resistant bacteria in piglets kept on a single farm. It demonstrated the presence of resistant strains of Staphylococcus spp., including one MRSA strain, Enterococcus spp. and Escherichia coli. These strains may be present as a result of postnatal colonization with both bacterial microflora of dams and environmental microflora.

  13. Being an Informed Consumer of Health Information and Assessment of Electronic Health Literacy in a National Sample of Internet Users: Validity and Reliability of the e-HLS Instrument.

    Science.gov (United States)

    Seçkin, Gül; Yeatts, Dale; Hughes, Susan; Hudson, Cassie; Bell, Valarie

    2016-07-11

    The Internet, with its capacity to provide information that transcends time and space barriers, continues to transform how people find and apply information to their own lives. With the current explosion in electronic sources of health information, including thousands of websites and hundreds of mobile phone health apps, electronic health literacy is gaining an increasing prominence in health and medical research. An important dimension of electronic health literacy is the ability to appraise the quality of information that will facilitate everyday health care decisions. Health information seekers explore their care options by gathering information from health websites, blogs, Web-based forums, social networking websites, and advertisements, despite the fact that information quality on the Internet varies greatly. Nonetheless, research has lagged behind in establishing multidimensional instruments, in part due to the evolving construct of health literacy itself. The purpose of this study was to examine psychometric properties of a new electronic health literacy (ehealth literacy) measure in a national sample of Internet users with specific attention to older users. Our paper is motivated by the fact that ehealth literacy is an underinvestigated area of inquiry. Our sample was drawn from a panel of more than 55,000 participants maintained by Knowledge Networks, the largest national probability-based research panel for Web-based surveys. We examined the factor structure of a 19-item electronic Health Literacy Scale (e-HLS) through exploratory factor analysis (EFA) and confirmatory factor analysis, internal consistency reliability, and construct validity on sample of adults (n=710) and a subsample of older adults (n=194). The AMOS graphics program 21.0 was used to construct a measurement model, linking latent factors obtained from EFA with 19 indicators to determine whether this factor structure achieved a good fit with our entire sample and the subsample (age ≥ 60

  14. Internal validation of the DNAscan/ANDE™ Rapid DNA Analysis™ platform and its associated PowerPlex(®) 16 high content DNA biochip cassette for use as an expert system with reference buccal swabs.

    Science.gov (United States)

    Moreno, Lilliana I; Brown, Alice L; Callaghan, Thomas F

    2017-07-01

    Rapid DNA platforms are fully integrated systems capable of producing and analyzing short tandem repeat (STR) profiles from reference sample buccal swabs in less than two hours. The technology requires minimal user interaction and experience making it possible for high quality profiles to be generated outside an accredited laboratory. The automated production of point of collection reference STR profiles could eliminate the time delay for shipment and analysis of arrestee samples at centralized laboratories. Furthermore, point of collection analysis would allow searching against profiles from unsolved crimes during the normal booking process once the infrastructure to immediately search the Combined DNA Index System (CODIS) database from the booking station is established. The DNAscan/ANDE™ Rapid DNA Analysis™ System developed by Network Biosystems was evaluated for robustness and reliability in the production of high quality reference STR profiles for database enrollment and searching applications. A total of 193 reference samples were assessed for concordance of the CODIS 13 loci. Studies to evaluate contamination, reproducibility, precision, stutter, peak height ratio, noise and sensitivity were also performed. The system proved to be robust, consistent and dependable. Results indicated an overall success rate of 75% for the 13 CODIS core loci and more importantly no incorrect calls were identified. The DNAscan/ANDE™ could be confidently used without human interaction in both laboratory and non-laboratory settings to generate reference profiles. Published by Elsevier B.V.

  15. Effects of gamma radiation and electron beam on samples of the Brazil nuts artificially inoculated with Aspergillus flavus; Efeitos da radiacao gama e feixe de eletrons sobre amostras de castanhas-do-Brasil inoculadas artificialmente com Aspergillus flavus

    Energy Technology Data Exchange (ETDEWEB)

    Coelho, Ednei Assuncao Antunes

    2012-07-01

    The high level of contamination by aflatoxin produced by fungi in lots of Brazil nuts and the strict control by importing countries in relation to the levels of toxins in food, European Union countries decided in 2003 by the return of these lots products from Brazil. Despite the economic loss represented by contamination by toxigenic fungi in Brazil nuts, a major product of extractive Northern of Brazil, studies are still preliminary as the control of contamination aflatoxigenic fungal using methods such as gamma radiation (G.R) and mainly, electron beam (E.B). These facts motivated this research, which aimed to evaluate the effects of gamma radiation and application of electron beam in samples of Brazil nut artificially inoculated with Aspergillus flavus. This goal, we were studied 50 samples of the Brazil nut previously inoculated with spores of A. flavus and subsequently incubated at 30 °C in relative humidity controlled at 93%. After incubation, period of 15 days, the average water activity of the samples was 0.80, the samples were divided into 5 groups that received the following doses of radiation: control (0 kGy), 5 and 10 kGy 5 E.B and G.R. The mycobiota was performed by serial dilution, plated on surface using potato dextrose agar. The results demonstrated that treatment with E.B using a dose of 5 kGy and 10 kGy resulted in reduced growth of A. flavus in 74% (37/50) and 94% (47/50) of samples. The samples treated with G.R at the dose of 5 kGy and 10 kGy no fungal growth occurred in 92% (46/50) 100% (50/50) of. The study of aflatoxins showed that doses of E.B of 5 kGy and 10 kGy reduced levels of AFB1 at 53.32% and 65.66% respectively. The application of gamma rays at doses of 5 and 10 kGy reduced levels of toxins in 70.61% and 84.15% respectively. This result may be attributed to higher penetrability of gamma radiation. Sensory analysis showed greater acceptance of the judges for the samples irradiated with E.B and G.R at the dose of 10 kGy. We concluded

  16. Use of eschar swabbing for the molecular diagnosis and genotyping of Orientia tsutsugamushi causing scrub typhus in Quang Nam province, Vietnam.

    Directory of Open Access Journals (Sweden)

    Nhiem Le Viet

    2017-02-01

    Full Text Available Scrub typhus is a rickettsiosis which is caused by Orientia tsutsugamushi and occurs throughout the Asia-Pacific region. Molecular diagnosis of rickettsioses using eschar swabs has recently emerged, and may be very useful for the diagnosis of these diseases in tropical settings.Quantitative polymerase chain reaction (qPCR was used to detect O. tsutsugamushi DNA in whole blood and eschar swab specimens of 67 patients who were clinically suspected of scrub typhus in Quang Nam province, Vietnam. Among the 20 patients for whom both eschar and whole blood were obtained, 17 (85% of the eschar specimens and 5 (25% of the whole blood specimens tested positive for O. tsutsugamushi. Genetic analysis of the 56-kDa TSA gene sequences demonstrated that the 14 sequences obtained in this study, including 12 eschar swabs and 2 whole blood specimens, were related to 4 groups: Karp, Kawasaki, Gilliam (JG-v and TG-v and TA716. The majority (9/14; 64.4% of contemporary O. tsutsugamushi genotypes in Quang Nam province were related to the Karp group.These results suggest that polyclonal antigen pools used for serological testing in the future should contain at least Karp, Kawasaki, Gilliam and TA716 antigens for Vietnamese patients, as well as patients who have traveled to Vietnam. qPCR after eschar swabbing should be considered for molecular diagnosis of scrub typhus in endemic patients as well as in travelers, since it is easy to perform and appears very useful for the rapid detection of Orientia tsutsugamushi in the early phase of infection.

  17. Eradication of methicillin-resistant Staphylococcus aureus in pressure ulcers comparing a polyhexanide-containing cellulose dressing with polyhexanide swabs in a prospective randomized study.

    Science.gov (United States)

    Wild, Thomas; Bruckner, Maria; Payrich, Martina; Schwarz, Christoph; Eberlein, Thomas; Andriessen, Anneke

    2012-01-01

    The study evaluated eradication of methicillin-resistant Staphylococcus aureus (MRSA) from pressure ulcers comparing swabs containing polyhexanide with a cellulose dressing + polyhexanide. After receiving approval from the ethics committee and informed consent, patients from the centers were recruited. Prospective randomized study. Thirty patients (n = 15/n = 15), not responding to wound disinfection after a washout period of 2 weeks, were included in the intention-to-treat analysis. This study was performed on hospital patients. Patients had pressure ulcers containing MRSA. For the control group, cleansing was performed with polyhexanide swabs (20 minutes), after which a foam dressing was applied. The study group received a polyhexanide-containing cellulose dressing. For bacterial analysis, semiquantitative swab cultures (Robert Koch Institute recommendations) were taken on days 0, 7, and 14 and during 3 consecutive days. The groups were comparable at baseline. At day 7, in the control group, 6 of 15 (40%) MRSA eradication. For the study group, there were 13 of 15 (86.67%) who showed MRSA eradication. At day 14, in the control group, there were 10 of 15 (66.67%) who had MRSA eradication, compared with the study group, where 15 of 15 (100%; P polyhexanide was shown to be successful in both groups, showing superior results for the study group.

  18. Development of a polymerase chain reaction to distinguish monocellate cobra (Naja khouthia) bites from other common Thai snake species, using both venom extracts and bite-site swabs.

    Science.gov (United States)

    Suntrarachun, S; Pakmanee, N; Tirawatnapong, T; Chanhome, L; Sitprija, V

    2001-07-01

    A PCR technique was used in this study to identify and distinguish monocellate cobra snake bites using snake venoms and swab specimens from snake bite-sites in mice from bites by other common Thai snakes. The sequences of nucleotide primers were selected for the cobrotoxin-encoding gene from the Chinese cobra (Naja atra) since the sequences of monocellate cobra (Naja kaouthia) venom are still unknown. However, the 113-bp fragment of cDNA of the cobrotoxin-encoding gene was detected in the monocellate cobra venom using RT-PCR. This gene was not found in the venoms of Ophiophagus hannah (king cobra), Bungarus fasciatus (banded krait), Daboia russelii siamensis (Siamese Russell's Viper, and Calloselasma rhodostoma (Malayan pit viper). Moreover, direct PCR could detect a 665-bp fragment of the cobrotoxin-encoding gene in the monocellate cobra venom but not the other snake venoms. Likewise, this gene was only observed in swab specimens from cobra snake bite-sites in mice. This is the first report demonstrating the ability of PCR to detect the cobrotoxin-encoding gene from snake venoms and swab specimens. Further studies are required for identification of this and other snakes from the bite-sites on human skin.

  19. Electronic detectors for electron microscopy.

    Science.gov (United States)

    Faruqi, A R; McMullan, G

    2011-08-01

    Electron microscopy (EM) is an important tool for high-resolution structure determination in applications ranging from condensed matter to biology. Electronic detectors are now used in most applications in EM as they offer convenience and immediate feedback that is not possible with film or image plates. The earliest forms of electronic detector used routinely in transmission electron microscopy (TEM) were charge coupled devices (CCDs) and for many applications these remain perfectly adequate. There are however applications, such as the study of radiation-sensitive biological samples, where film is still used and improved detectors would be of great value. The emphasis in this review is therefore on detectors for use in such applications. Two of the most promising candidates for improved detection are: monolithic active pixel sensors (MAPS) and hybrid pixel detectors (of which Medipix2 was chosen for this study). From the studies described in this review, a back-thinned MAPS detector appears well suited to replace film in for the study of radiation-sensitive samples at 300 keV, while Medipix2 is suited to use at lower energies and especially in situations with very low count rates. The performance of a detector depends on the energy of electrons to be recorded, which in turn is dependent on the application it is being used for; results are described for a wide range of electron energies ranging from 40 to 300 keV. The basic properties of detectors are discussed in terms of their modulation transfer function (MTF) and detective quantum efficiency (DQE) as a function of spatial frequency.

  20. Comparison of noninvasive sample collection procedures for canine leishmaniasis diagnosis by PCR-hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, Sidney de Almeida; Andrade, Antero Silva Ribeiro de [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)]. E-mail: vidasnino@yahoo.com.br; antero@cdtn.br; Ituassu, Leonardo Trindade; Melo, Maria Norma de [Universidade Federal de Minas Gerais, Belo Horizonte, MG (Brazil)]. E-mail: melo@mono.icb.ufmg.br; ltituassu@yahoo.com.br

    2007-07-01

    The dogs are the main reservoir of the visceral leishmaniasis etiological agent Leishmania chagasi and these animals have to be systematically monitored. The aim of present work was to standardize a method for canine leishmaniasis diagnosis using DNA samples obtained by a noninvasive ways. Two kind of samples were compared: conjunctival swab and blood. The samples were analyzed by the Polymerase Chain Reaction (PCR) associated with the hybridization of {sup 32}P labeled DNA probes. An in vitro test was carried out using cotton swabs seeded with L. chagasi parasites at different cell numbers. After that, the PCR and hybridization sensitivity was evaluated in two groups of 23 seropositive dogs. Conjunctival swabs and 1,0 mL of blood were collected from each animal. 90 {mu}L of these blood were spotted onto filter paper and the remaining used to prepare the buffy coat. The DNA purification from cotton swabs was carried out through the phenol-chloroform (group 1) or boiling (group 2). The Wizard kit was used to DNA extraction from buffy coat. The filters were treated according to Dialab protocol. The analysis of the seeded samples showed that the PCR was able to identify until ten parasites while the following hybridization of the PCR products allows the detection of until one parasite. The PCR positivity for the conjunctival swabs were 73.9% and 52.2% respectively to the groups 1 and 2. For buffy coat the positivities were 43.5% and 56.5% respectively. The filters presented the lowest positivity. The hybridization step was not accomplished yet for these samples. (author)

  1. Comparison of sampling procedures and microbiological and non-microbiological parameters to evaluate cleaning and disinfection in broiler houses.

    Science.gov (United States)

    Luyckx, K; Dewulf, J; Van Weyenberg, S; Herman, L; Zoons, J; Vervaet, E; Heyndrickx, M; De Reu, K

    2015-04-01

    Cleaning and disinfection of the broiler stable environment is an essential part of farm hygiene management. Adequate cleaning and disinfection is essential for prevention and control of animal diseases and zoonoses. The goal of this study was to shed light on the dynamics of microbiological and non-microbiological parameters during the successive steps of cleaning and disinfection and to select the most suitable sampling methods and parameters to evaluate cleaning and disinfection in broiler houses. The effectiveness of cleaning and disinfection protocols was measured in six broiler houses on two farms through visual inspection, adenosine triphosphate hygiene monitoring and microbiological analyses. Samples were taken at three time points: 1) before cleaning, 2) after cleaning, and 3) after disinfection. Before cleaning and after disinfection, air samples were taken in addition to agar contact plates and swab samples taken from various sampling points for enumeration of total aerobic flora, Enterococcus spp., and Escherichia coli and the detection of E. coli and Salmonella. After cleaning, air samples, swab samples, and adenosine triphosphate swabs were taken and a visual score was also assigned for each sampling point. The mean total aerobic flora determined by swab samples decreased from 7.7±1.4 to 5.7±1.2 log CFU/625 cm2 after cleaning and to 4.2±1.6 log CFU/625 cm2 after disinfection. Agar contact plates were used as the standard for evaluating cleaning and disinfection, but in this study they were found to be less suitable than swabs for enumeration. In addition to measuring total aerobic flora, Enterococcus spp. seemed to be a better hygiene indicator to evaluate cleaning and disinfection protocols than E. coli. All stables were Salmonella negative, but the detection of its indicator organism E. coli provided additional information for evaluating cleaning and disinfection protocols. Adenosine triphosphate analyses gave additional information about the

  2. Comparison of sample types and diagnostic methods for in vivo detection of Mycoplasma hyopneumoniae during early stages of infection.

    Science.gov (United States)

    Pieters, Maria; Daniels, Jason; Rovira, Albert

    2017-05-01

    Detection of Mycoplasma hyopneumoniae in live pigs during the early stages of infection is critical for timely implementation of control measures, but is technically challenging. This study compared the sensitivity of various sample types and diagnostic methods for detection of M. hyopneumoniae during the first 28days after experimental exposure. Twenty-one 8-week old pigs were intra-tracheally inoculated on day 0 with M. hyopneumoniae strain 232. Two age matched pigs were mock inoculated and maintained as negative controls. On post-inoculation days 0, 2, 5, 9, 14, 21 and 28, nasal swabs, laryngeal swabs, tracheobronchial lavage fluid, and blood samples were obtained from each pig and oral fluid samples were obtained from each room in which pigs were housed. Serum samples were assayed by ELISA for IgM and IgG M. hyopneumoniae antibodies and C-reactive protein. All other samples were tested for M. hyopneumoniae DNA by species-specific real-time PCR. Serum antibodies (IgG) to M. hyopneumoniae were detected in challenge-inoculated pigs on days 21 and 28. M. hyopneumoniae DNA was detected in samples from experimentally inoculated pigs beginning at 5days post-inoculation. Laryngeal swabs at all samplings beginning on day 5 showed the highest sensitivity for M. hyopneumoniae DNA Detection, while oral fluids showed the lowest sensitivity. Although laryngeal swabs are not considered the typical M. hyopneumoniae diagnostic sample, under the conditions of this study laryngeal swabs tested by PCR proved to be a practical and reliable diagnostic sample for M. hyopneumoniae detection in vivo during early-stage infection. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Application of modified hollow fiber liquid phase microextraction in conjunction with chromatography-electron capture detection for quantification of acrylamide in waste water samples at ultra-trace levels.

    Science.gov (United States)

    Sobhi, Hamid Reza; Ghambarian, Mahnaz; Behbahani, Mohammad; Esrafili, Ali

    2017-03-03

    Herein, a simple and sensitive method was successfully developed for the extraction and quantification of acrylamide in water samples. Initially, acrylamide was derivatized through a bromination process. Subsequently, a modified hollow-fiber liquid-phase microextraction was applied for the extraction of the brominated acrylamide from a 10-ml portion of an aqueous sample. Briefly, in this method, the derivatized acrylamide (2,3-dibromopropionamide) was extracted from the aqueous sample into a thin layer of an organic solvent sustained in pores of a porous hollow fiber. Then, it was back-extracted using a small volume of organic acceptor solution (acetonitril, 25μl) located inside the lumen of the hollow fiber followed by gas chromatography-electron capture detection (GC-ECD). The optimal conditions were examined for the extraction of the analyte such as: the organic solvent: dihexyl ether+10% tri-n-octyl phosphine oxide; stirring rate: 750rpm; no salt addition and 30min extraction time. These optimal extraction conditions allowed excellent enrichment factor values for the method. Enrichment factor, detection limit (S/N=3) and dynamic linear range of 60, 2ngL-1 and 50-1000ngL-1 to be determined for the analyte. The relative standard deviations (RSD%) representing precision of the method were in the range of 2.2-5.8 based on the average of three measurements. Accuracy of the method was tested by the relative recovery experiments on spiked samples, with results ranging from 93 to 108%. Finally, the method proved to be simple, rapid, and cost-effective for routine screen of acrylamide-contaminated highly-complicated untreated waste water samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Performance of four different agar plate methods for rectal swabs, synergy disk tests and metallo-β-lactamase Etest for clinical isolates in detecting carbapenemase-producing Klebsiella pneumoniae.

    Science.gov (United States)

    Papadimitriou-Olivgeris, Matthaios; Vamvakopoulou, Sophia; Spyropoulou, Αikaterini; Bartzavali, Christina; Marangos, Markos; Anastassiou, Evangelos D; Spiliopoulou, Iris; Christofidou, Myrto

    2016-09-01

    The aims of the study were to compare four different agar plate methods in the identification of carbapenemase-producing Klebsiella pneumoniae (CP-Kp) from rectal samples and to assess the role of phenotypic methodologies in the identification of carbapenemase type from clinical K. pneumoniae isolates. Two chromogenic agars (Brilliance CRE and CHROMagar KPC) were compared to MacConkey agar plates with ertapenem (ERT) or imipenem (IMP) disks for the identification of CP-Kp from 912 rectal swabs. CP-Kp was detected in 329 samples by either agar methodology (299 K. pneumoniae carbapenemase positive, 27 Verona integron-encoded metallo-β-lactamase positive and 3 K. pneumoniae carbapenemase and Verona integron-encodedmetallo-β-lactamase positive). Sensitivity of Brilliance CRE, CHROMagar KPC and MacConkey agar plus IMP or ERT disk (inhibition zone 97.5 %). Phenotypic methodologies can provide reliable results for the identification of carbapenemase production among K. pneumoniae isolates. Chromogenic agars can be applied in high-risk patients as part of surveillance and infection control programs.

  5. A Novel Method for Profiling and Quantifying Short- and Medium-Chain Chlorinated Paraffins in Environmental Samples Using Comprehensive Two-Dimensional Gas Chromatography-Electron Capture Negative Ionization High-Resolution Time-of-Flight Mass Spectrometry.

    Science.gov (United States)

    Xia, Dan; Gao, Lirong; Zheng, Minghui; Tian, Qichang; Huang, Huiting; Qiao, Lin

    2016-07-19

    Chlorinated paraffins (CPs) are complex technical mixtures containing thousands of isomers. Analyzing CPs in environmental matrices is extremely challenging. CPs have broad, unresolved profiles when analyzed by one-dimensional gas chromatography (GC). Comprehensive two-dimensional GC (GC×GC) can separate CPs with a high degree of orthogonality. A novel method for simultaneously profiling and quantifying short- and medium-chain CPs, using GC×GC coupled with electron capture negative ionization high-resolution time-of-flight mass spectrometry, was developed. The method allowed 48 CP formula congener groups to be analyzed highly selectively in one injection through accurate mass measurements of the [M - Cl](-) ions in full scan mode. The correlation coefficients (R(2)) for the linear calibration curves for different chlorine contents were 0.982 for short-chain CPs and 0.945 for medium-chain CPs. The method was successfully used to determine CPs in sediment and fish samples. By using this method, with enhanced chromatographic separation and high mass resolution, interferences between CP congeners and other organohalogen compounds, such as toxaphene, are minimized. New compounds, with the formulas C9H14Cl6 and C9H13Cl7, were found in sediment and biological samples for the first time. The method was shown to be a powerful tool for the analysis of CPs in environmental samples.

  6. Microbiological criteria in public catering: sampling and auditing experiences in canteens and cafeterias in Piedmont

    Directory of Open Access Journals (Sweden)

    Amaranta Traversa

    2013-02-01

    Full Text Available In the period 2006-2011 six public catering establishments (3 canteens and 3 cafeterias were monitored, trough audit and sampling, in order to verify the application of good manufacturing and hygiene practices during food production, handling and serving. The compliance to microbiological food safety criteria (Listeria monocytogenes and Salmonella spp. and process hygiene criteria were investigated using ISO standards for microbiological analyses. A total of 612 samples were collected: 192 food samples and 288 environmental swab samples from canteens; 33 food samples and 99 swab samples from cafeterias. Regarding food safety, two samples were in disagreement with criteria fixed in EU Regulation as Listeria monocytogenes was isolated from a turkey breast in a canteen and from a sandwich in a cafeteria. Regarding process hygiene criteria, as no microbiological limits are legally defined for catering services, for this study limits were fixed according to the quality standards of tender, scientific literature and laboratory experience. 23.4% foodstuffs and 8.7% swabs resulted non-compliant in canteens; 48.5% foodstuffs and 6.1% swabs resulted non-compliant in cafeterias. The count of coagulase-positive staphylococci (CPS was higher of the fixed limits in raw turkey meat and in cooked spinach: the presence among CPS of S. aureus was confirmed, strains were not able to produce enterotoxins. The most common non-compliance in hygiene criteria was represented by aerobic colony count (60.7% of total non-compliance in canteens and 75.0% in cafeterias and coliform bacteria (20.3% in canteens and 25.0% in cafeterias. Nine raw foods or raw readyto- eat food samples were non-compliant for both coliform bacteria and aerobic count; one sample (raw turkey meat was non-compliant for CPS and aerobic count but resulted to be compliant after cooking. Auditing and sampling are the most effective tools to improve food quality standard and to enhance food business

  7. DISTRIBUTION OF COAGULASE-NEGATIVE STAPHYLOCOCCI SPECIES ON TEAT SKIN AND IN MILK SAMPLES FROM DAIRY COWS IN AUTOMATIC MILKING SYSTEMS

    DEFF Research Database (Denmark)

    Mahmmod, Yasser Saadeldien Ibrahim; Svennesen, Line; Pedersen, Karl

    Coagulase-negative staphylococci (CNS) frequently colonize teat skin and are common findings cultured from milk samples of cows with subclinical intramammary infections (IMI). Several species are related to IMI, but knowledge about the epidemiology of CNS species is limited. Cows in automatic...... milking systems (AMS) may have increased risk for teat colonization and IMI because more than 60 cows are milked several times daily with the same milking unit. The objectives of this study are (1) to investigate patterns of CNS species in milk samples and teat skin swabs in nine AMS herds and (2......) to identify the predisposing cow level risk factors for specific CNS IMI and teat colonization.In each herd, 30- 40 cows with somatic cell counts > 200,000 cells/ml in the previous milk recording are randomly selected and teat skin swabs and aseptic quarter foremilk samples are taken. Teat skin swabs...

  8. Comparison of Gas Chromatography-Mass Spectrometry and Gas Chromatography-Tandem Mass Spectrometry with Electron Ionization and Negative-Ion Chemical Ionization for Analyses of Pesticides at Trace Levels in Atmospheric Samples

    Science.gov (United States)

    Raina, Renata; Hall, Patricia

    2008-01-01

    A comparison of detection limits of gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) with gas chromatography-tandem mass spectrometry (GC-MS/MS) in selected reaction monitoring (SRM) mode with both electron ionization (EI) and negative-ion chemical ionization (NCI) are presented for over 50 pesticides ranging from organochlorines (OCs), organophosphorus pesticides (OPs) and pre-emergent herbicides used in the Canadian prairies (triallate, trifluralin, ethalfluralin). The developed GC-EI/SIM, GC-NCI/SIM, and GC-NCI/SRM are suitable for the determination of pesticides in air sample extracts at concentrations ethion, and OCs: alachlor, aldrin, perthane, and DDE, DDD, DDT). PMID:19609395

  9. Comparison of the Cepheid GeneXpert CT/NG assay to the Hologic Aptima Combo2 assay for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in self-collected rectal swabs.

    Science.gov (United States)

    Dize, Laura; Silver, Barbara; Gaydos, Charlotte

    2018-02-01

    Self-collected rectal-swabs were tested for CT and NG on GeneXpert CT/NG as compared to APTIMA Combo2. Of 448 rectal-swabs, 22 were positive for CT; 7 for NG on both assays; two were discordant. Sensitivity and specificity of GeneXpert was 95.5% and 99.7% for chlamydia, respectively; for gonorrhea both were 100%. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Ultra-reduced phases in Apollo 16 regolith: Combined field emission electron probe microanalysis and atom probe tomography of submicron Fe-Si grains in Apollo 16 sample 61500

    Science.gov (United States)

    Gopon, Phillip; Spicuzza, Michael J.; Kelly, Thomas F.; Reinhard, David; Prosa, Ty J.; Fournelle, John

    2017-09-01

    The lunar regolith contains a variety of chemically reduced phases of interest to planetary scientists and the most common, metallic iron, is generally ascribed to space weathering processes (Lucey et al. ). Reports of silicon metal and iron silicides, phases indicative of extremely reducing conditions, in lunar samples are rare (Anand et al. ; Spicuzza et al. ). Additional examples of Fe-silicides have been identified in a survey of particles from Apollo 16 sample 61501,22. Herein is demonstrated the utility of low keV electron probe microanalysis (EPMA), using the Fe Ll X-ray line, to analyze these submicron phases, and the necessity of accounting for carbon contamination. We document four Fe-Si and Si0 minerals in lunar regolith return material. The new Fe-Si samples have a composition close to (Fe,Ni)3Si, whereas those associated with Si0 are close to FeSi2 and Fe3Si7. Atom probe tomography of (Fe,Ni)3Si shows trace levels of C (60 ppma and nanodomains enriched in C, Ni, P, Cr, and Sr). These reduced minerals require orders of magnitude lower oxygen fugacity and more reducing conditions than required to form Fe0. Documenting the similarities and differences in these samples is important to constrain their formation processes. These phases potentially formed at high temperatures resulting from a meteorite impact. Whether carbon played a role in achieving the lower oxygen fugacities—and there is evidence of nearby carbonaceous chondritic material—it remains to be proven that carbon was the necessary component for the unique existence of these Si0 and iron silicide minerals.

  11. [Comparison of the detection rate and load of rhinovirus in nasal versus lower airway samples from patients with chronic obstructive pulmonary disease].

    Science.gov (United States)

    Liao, Hua; Chen, Rongchang; Guan, Wenda; Yang, Zifeng; Liu, Wenkuan; Zhou, Rong; Zhong, Nanshan

    2014-01-01

    To compare the rhinovirus detection rate and the viral load in nasal samples versus lower airway samples from patients with COPD, and therefore to provide evidence for sampling selection for detection of rhinovirus. Nasal swab and induced sputum were collected from patients with COPD during acute exacerbation and the stable period. Rhinovirus was detected by real-time fluorescent quantitative polymerase chain reaction. The difference in detection rates of rhinovirus between acute exacerbation and stable COPD was compared. The detection rates and the viral load from nasal samples versus induced sputum were also compared. A total of 639 paired nasal swab and induced sputum specimens were collected between September 2009 and January 2013, including 114 paired specimens from COPD patients with acute exacerbations 114 paired specimens from stable COPD (matching with the stable one), and 411 paired specimens from stable COPD patients. For the 114 paired samples from stable and acute COPD patients, there was a higher detection rate in samples [nasal swab 13.2% (15/114) , induced sputum 21.9% (25/114) ] from patients with acute exacerbation, compared those with stable disease [nasal swab 3.5% (4/114), P rhinovirus, while 58(9.1%) of the 639 induced sputum specimens were positive for rhinovirus (P rhinovirus positive specimens, the mean viral load of rhinovirus in induced sputum was (62.1 ± 9.5) × 10⁸ copies/L, significantly higher than that of the nasal swab (3.4 ± 0.5) × 10⁸ copies/L, P rhinovirus detection compared to nasal swabs, and the load of rhinovirus was higher in the lower airways than in the upper airways.

  12. Rapid determination of dichlorodiphenyltrichloroethane and its main metabolites in aqueous samples by one-step microwave-assisted headspace controlled-temperature liquid-phase microextraction and gas chromatography with electron capture detection.

    Science.gov (United States)

    Vinoth Kumar, Ponnusamy; Jen, Jen-Fon

    2011-03-01

    A rapid and sensitive analytical method for the determination of dichlorodiphenyltrichloroethane (DDT) and its main metabolites in environmental aqueous samples has been developed using one-step microwave-assisted headspace controlled-temperature liquid-phase micro-extraction (MA-HS-CT-LPME) technique coupled with gas chromatography-electron-capture detection (GC-ECD). In this study, the one-step extraction of DDT and its main metabolites was achieved by using microwave heating to accelerate the evaporation of analytes into the controlled-temperature headspace to form a cloudy mist vapor zone for LPME sampling. Parameters influencing extraction efficiency were thoroughly optimized, and the best extraction for DDT and its main metabolites from 10-mL aqueous sample at pH 6.0 was achieved by using 1-octanol (4-μL) as the LPME solvent, sampling at 34°C for 6.5 min under 249W of microwave irradiation. Under optimum conditions, excellent linear relationship was obtained in the range of 0.05-1.0 μg/L for 1-dichloro-2,2-bis-(p'-chlorophenyl)ethylene (p,p'-DDE), 0.1-2.0 μg/L for o,p'-DDT, 0.15-3.0 μg/L for 1,1-dichloro-2,2-bis-(p'-chlorophenyl)ethane (p,p'-DDD) and p,p'-DDT, with detection limits of 20 ng/L for p,p'-DDE, and 30 ng/L for o,p'-DDT, p,p'-DDD and p,p'-DDT. Precision was in the range of 3.2-11.3% RSD. The proposed method was validated with environmental water samples. The spiked recovery was between 95.5% and 101.3% for agricultural-field water, between 94% and 99.7% for sea water and between 93.5% and 98% for river water. Thus the established method has been proved to be a simple, rapid, sensitive, inexpensive and eco-friendly procedure for the determination of DDT and its main metabolites in environmental water samples. Copyright © 2010 Elsevier Ltd. All rights reserved.

  13. Simultaneous analysis of non-steroidal anti-inflammatory drugs and estrogenic hormones in water and wastewater samples using gas chromatography-mass spectrometry and gas chromatography with electron capture detection

    Energy Technology Data Exchange (ETDEWEB)

    Migowska, Natalia; Caban, Magda; Stepnowski, Piotr; Kumirska, Jolanta, E-mail: kumirska@chem.univ.gda.pl

    2012-12-15

    Non-steroidal anti-inflammatory drugs are the group of pharmaceuticals that is most often found in the environment, whereas estrogenic hormones are considered to be potent endocrine disruptors. However, the fate and persistence of these compounds in the environment are still unclear. In this study we propose two approaches for determining these compounds in environmental water samples: GC-MS using time windows and operating in selected ion-monitoring mode (SIM) and, for the first time, gas chromatography with electron capture detection (GC-ECD). The identification criteria of both methods fulfilled the requirements of Directive 2002/657/EC. The use of time windows improved the sensitivity of GC-MS measurements. In GC-MS analysis the pharmaceuticals were determined as trimethylsilyl, in GC-ECD as pentafluoropropionyl derivatives. The influence of such parameters as the type of reagent, type of solvent, reaction time, reaction temperature and microwave irradiation in a household microwave oven on the efficacy of silylation was investigated. Derivatization using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and 1% trimethylchlorosilane (TMCS) in pyridine (1:1, v/v) for 30 min in 60 Degree-Sign C was found to be optimal. Optimization of the solid phase extraction procedure (SPE) confirmed that the application of Oasis HLB cartridges, the acidification of loading samples to pH 2 and the use of methanol as eluent gave the best absolute recoveries (ARs) of the target compounds. The following ARs of all the compounds were achieved: 58.2-106.8% in influent wastewater, 77.8-103.4% in effluent wastewater and 81.2-101.9% in surface water samples. Validation of the SPE-GC-MS method enables 13 pharmaceuticals to be determined with MDLs between 3.3 and 343.6 ng/L, depending on the analytes and matrices. GC-ECD analysis enables the determination of 6 pharmaceuticals in surface water samples with MDLs between 0.7 and 5.4 ng/L. The proposed methods were successfully used for

  14. Destructive and nondestructive procedures to obtain chicken carcass samples for Escherichia coli and Salmonella spp. detection.

    Science.gov (United States)

    Cossi, Marcus Vinícius Coutinho; de Almeida, Michelle Vieira; Dias, Mariane Rezende; de Arruda Pinto, Paulo Sérgio; Nero, Luís Augusto

    2011-12-01

    Destructive and nondestructive sampling procedures were compared for Escherichia coli and Salmonella spp. detection in 60 fresh chicken carcasses, which were submitted to the following sampling procedures: rinsing, skin swabbing, tissue excision, and skin excision; the proximity or not to the cloacae region was also considered. The obtained results were compared to identify significant differences (pchicken carcasses were positive for E. coli, and five were positive for Salmonella spp. For E. coli, nonsignificant differences were observed between rinsing and tissue excision, rinsing and skin excision, and skin excision and tissue excision (p>0.05), thus indicating equivalencies between these techniques. Skin swabbing produced a statistically significant lower frequency of positive results (pE. coli, thus indicating its inadequacy for detection of this microorganism. For Salmonella spp., no significant differences were observed between the sampling techniques (p>0.05), possibly due to the low overall frequency of positive carcasses. No significant differences in the number of positive samples (E. coli or Salmonella spp.) were observed between samples collected near or far from the cloacae region (p>0.05), regardless of the sampling technique. The obtained results demonstrate that the tested sampling techniques were equivalent for Salmonella spp. detection in chicken carcasses, as observed for E. coli with the exception of skin swabbing.

  15. Identification of a novel human papillomavirus by metagenomic analysis of samples from patients with febrile respiratory illness

    NARCIS (Netherlands)

    Mokili, J.L.; Dutilh, B.E.; Lim, Y.W.; Schneider, B.S.; Taylor, T.; Haynes, M.R.; Metzgar, D.; Myers, C.A.; Blair, P.J.; Nosrat, B.; Wolfe, N.D.; Rohwer, F.

    2013-01-01

    As part of a virus discovery investigation using a metagenomic approach, a highly divergent novel Human papillomavirus type was identified in pooled convenience nasal/oropharyngeal swab samples collected from patients with febrile respiratory illness. Phylogenetic analysis of the whole genome and

  16. Comparison of nasopharyngeal aspirate and nasal swab specimens for detection of respiratory syncytial virus in different settings in a developing country

    DEFF Research Database (Denmark)

    Stensballe, L G; Trautner, S; Kofoed, P-E

    2002-01-01

    OBJECTIVE: To compare detection of respiratory syncytial virus (RSV) for diagnostic purposes using nasopharyngeal aspirate (NPA) and nasal swabs (NS) in different clinical settings in a community study in Guinea-Bissau. METHOD: During 1996-98 paired specimens were obtained from 635 children under 5...... years of age (median: 274 days; interquartile range: 144-453 days) with symptoms of lower respiratory infections (LRI). The specimens were analysed by an enzyme-linked immunosorbent assay for RSV antigen in Guinea-Bissau and re-analysed in Denmark using the same assay. The gold standard for RSV antigen...

  17. Anatomical characteristics of teats and premilking bacterial counts of teat skin swabs of primiparous cows exposed to different types of bedding.

    Science.gov (United States)

    Guarín, J F; Baumberger, C; Ruegg, P L

    2017-02-01

    Bacterial populations of teat skin are associated with risk of intramammary infection and may be influenced by anatomical characteristics of teats. The objective of this study was to evaluate associations of selected anatomical characteristics of teats with bacterial counts of teat skin of cows exposed to different types of bedding. Primarily primiparous Holstein cows (n = 128) were randomly allocated to 4 pens within a single barn. Each pen contained 1 type of bedding [new sand (NES), recycled sand (RS), deep-bedded manure solids (DBMS), and shallow-bedded manure solids over foam core mattresses (SBMS)]. During a single farm visit udders (n = 112) were scored for hygiene and 1 front (n = 112) and 1 rear teat (n = 111) of each enrolled cow were scored for hyperkeratosis (HK). Teat length, teat barrel diameter, and teat apex diameter were measured and teat skin swabs were systematically collected for microbiological analysis. Linear type evaluation data for udders of each cow were retrieved for each cow. Teat position (front or rear) was associated with occurrence of clinical mastitis during the 12 mo before the farm visit and more cases occurred in front quarters. The proportion of udders that were classified as clean (score 1 or 2) was 68, 82, 54, and 95% for cows housed in pens containing NES, RS, SBMS, and DBMS, respectively. No association was found between HK score and teat position and no association was found between HK score and teat skin bacterial count. Bacterial counts of teat skin swabs from front teats of cows in pens containing RS and SBMS were significantly less than those of rear teats of cows in pens containing DBMS or NES. Teat skin bacterial counts were significantly greater for swabs obtained from teats of cows with udder hygiene scores of 3 and 4 as compared with swabs obtained from cows with cleaner udders. Of all udder conformation traits evaluated, only narrower rear teat placement was positively associated with bacterial counts on teat skin

  18. Identification of Streptococcus parasanguinis DNA contamination in human buccal DNA samples

    OpenAIRE

    Mahfuz, Istiak; Cheng, Wei; White, Stefan J

    2013-01-01

    Background The use of buccal swabs in clinical and scientific studies is a very popular method of collecting DNA, due to its non-invasive nature of collection. However, contamination of the DNA sample may interfere with analysis. Findings Here we report the finding of Streptococcus parasanguinis bacterial DNA contamination in human buccal DNA samples, which led to preferential amplification of bacterial sequence with PCR primers designed against human sequence. Conclusion Contamination of buc...

  19. Inter-laboratory and inter-assay comparison on two real-time PCR techniques for quantification of PCV2 nucleic acid extracted from field samples

    DEFF Research Database (Denmark)

    Hjulsager, Charlotte Kristiane; Grau-Roma, L.; Sibila, M.

    2009-01-01

    linear association between the assays (p sample (p ... association between the amount of PCV2 DNA and the amount of total DNA, neither in nasal (p = 0.86) nor in rectal (p=0.78) swabs, suggesting that normalizing of PCV2 DNA load in swab samples to total DNA concentration is not suitable. The present exploratory study highlights the need for the performance...... of ring trials on qPCV2 protocols between laboratories. Meanwhile, the proposed thresholds for PMWS diagnosis should only be considered reliable for each particular laboratory and each particular assay....

  20. Evaluation of Two Surface Sampling Methods for Microbiological and Chemical Analyses To Assess the Presence of Biofilms in Food Companies.

    Science.gov (United States)

    Maes, Sharon; Huu, Son Nguyen; Heyndrickx, Marc; Weyenberg, Stephanie van; Steenackers, Hans; Verplaetse, Alex; Vackier, Thijs; Sampers, Imca; Raes, Katleen; Reu, Koen De

    2017-12-01

    Biofilms are an important source of contamination in food companies, yet the composition of biofilms in practice is still mostly unknown. The chemical and microbiological characterization of surface samples taken after cleaning and disinfection is very important to distinguish free-living bacteria from the attached bacteria in biofilms. In this study, sampling methods that are potentially useful for both chemical and microbiological analyses of surface samples were evaluated. In the manufacturing facilities of eight Belgian food companies, surfaces were sampled after cleaning and disinfection using two sampling methods: the scraper-flocked swab method and the sponge stick method. Microbiological and chemical analyses were performed on these samples to evaluate the suitability of the sampling methods for the quantification of extracellular polymeric substance components and microorganisms originating from biofilms in these facilities. The scraper-flocked swab method was most suitable for chemical analyses of the samples because the material in these swabs did not interfere with determination of the chemical components. For microbiological enumerations, the sponge stick method was slightly but not significantly more effective than the scraper-flocked swab method. In all but one of the facilities, at least 20% of the sampled surfaces had more than 102 CFU/100 cm2. Proteins were found in 20% of the chemically analyzed surface samples, and carbohydrates and uronic acids were found in 15 and 8% of the samples, respectively. When chemical and microbiological results were combined, 17% of the sampled surfaces were contaminated with both microorganisms and at least one of the analyzed chemical components; thus, these surfaces were characterized as carrying biofilm. Overall, microbiological contamination in the food industry is highly variable by food sector and even within a facility at various sampling points and sampling times.

  1. The Electron

    Energy Technology Data Exchange (ETDEWEB)

    Thomson, George

    1972-01-01

    Electrons are elementary particles of atoms that revolve around and outside the nucleus and have a negative charge. This booklet discusses how electrons relate to electricity, some applications of electrons, electrons as waves, electrons in atoms and solids, the electron microscope, among other things.

  2. Enhanced performance of methamphetamine lateral flow cassettes using an electronic lateral flow reader.

    Science.gov (United States)

    Smith, Jerome P; Sammons, Deborah L; Robertson, Shirley A; Snawder, John E

    2015-01-01

    Surface contamination from methamphetamine in meth labs continues to be a problem. We had previously developed a lateral flow assay cassette for field detection of methamphetamine contamination that is commercially available and has been used by a number of groups to assess contamination. This cassette uses the complete disappearance of the test line as an end point for detection of 50 ng/100 cm2 of methamphetamine contamination for surface sampling with cotton swabs. In the present study, we further evaluate the response of the cassettes using an electronic lateral flow reader to measure the intensities of the test and control lines. The cassettes were capable of detecting 0.25 ng/ml for calibration solutions. For 100 cm2 ceramic tiles that were spiked with methamphetamine and wiped with cotton-tipped wooden swabs wetted in assay/sampling buffer, 1 ng/tile was detected using the reader. Semi-quantitative results can be produced over the range 0-10 ng/ml for calibration solutions and 0-25 ng/tile for spiked tiles using either a 4-parameter logistic fit of test line intensity versus concentration or spiked mass or the ratio of the control line to the test line intensity fit to concentration or spiked mass. Recovery from the tiles was determined to be about 30% using the fitted curves. Comparison of the control line to the test line was also examined as a possible visual detection end point and it was found that the control line became more intense than the test line at 0.5 to 1 ng/ml for calibration solutions or 1 to 2 ng/tile for spiked tiles. Thus the lateral flow cassettes for methamphetamine have the potential to produce more sensitive semi-quantitative results if an electronic lateral flow reader is used and can be more sensitive for detection if the comparison of the control line to the test line is used as the visual end point.

  3. Primary secondary amine as a sorbent material in dispersive solid-phase extraction clean-up for the determination of indicator polychlorinated biphenyls in environmental water samples by gas chromatography with electron capture detection.

    Science.gov (United States)

    Guo, Yuanming; Hu, Hongmei; Li, Tiejun; Xue, Lijian; Zhang, Xiaoning; Zhong, Zhi; Zhang, Yurong; Jin, Yanjian

    2017-08-01

    A simple, rapid, and novel method has been developed and validated for determination of seven indicator polychlorinated biphenyls in water samples by gas chromatography with electron capture detection. 1 L of water samples containing 30 g of anhydrous sodium sulfate was first liquid-liquid extracted with an automated Jipad-6XB vertical oscillator using n-hexane/dichloromethane (1:1, v/v). The concentrated extract was cleaned up by dispersive solid-phase extraction with 100 mg of primary secondary amine as sorbent material. The linearity of this method ranged from 1.25 to 100 μg/L, with regression coefficients ranging between 0.9994 and 0.9999. The limits of detection were in the ng/L level, ranging between 0.2 and 0.3 ng/L. The recoveries of seven spiked polychlorinated biphenyls with external calibration method at different concentration levels in tap water, lake water, and sea water were in the ranges of 85-112, 76-116, and 72-108%, respectively, and with relative standard deviations of 3.3-4.5, 3.4-5.6, and 3.1-4.8% (n = 5), respectively. The performance of the proposed method was compared with traditional liquid-liquid extraction and solid-phase extraction clean-up methods, and comparable efficiencies were obtained. It is concluded that this method can be successfully applied for the determination of polychlorinated biphenyls in different water samples. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Impact of electronic faucets and water quality on the occurrence of Pseudomonas aeruginosa in water: a multi-hospital study.

    Science.gov (United States)

    Charron, Dominique; Bédard, Emilie; Lalancette, Cindy; Laferrière, Céline; Prévost, Michèle

    2015-03-01

    To compare Pseudomonas aeruginosa prevalence in electronic and manual faucets and assess the influence of connecting pipes and water quality. Faucets in 4 healthcare centers in Quebec, Canada. Water samples from 105 electronic, 90 manual, and 14 foot-operated faucets were analyzed for P. aeruginosa by culture and enzymatic detection, and swab samples from drains and aerators were analyzed by culture. Copper and residual chlorine concentrations, temperature, and flow rate were measured. P. aeruginosa concentrations were analyzed in 4 consecutive volumes of cold water and a laboratory study was conducted on copper pipes and flexible hoses. P. aeruginosa contamination was found in drains more frequently (51%) than in aerators (1%) or water (culture: 4%, enzyme detection: 16%). Prevalence in water samples was comparable between manual (14%) and 2 types of electronic faucets (16%) while higher for foot-operated faucets (29%). However, type 2 electronic faucets were more often contaminated (31%) than type 1 (14%), suggesting that faucet architecture and mitigated volume (30 mL vs 10 mL) influence P. aeruginosa growth. Concentrations were 100 times higher in the first 250 mL than after flushing. Flexible hoses were more favorable to P. aeruginosa growth than copper and a temperature of 40°C led to higher counts. The types of faucets and connecting pipes, flow rate, and water quality are important parameters influencing the prevalence and the concentrations of P. aeruginosa in faucets. High concentrations of P. aeruginosa in the first 250 mL suggest increased risk of exposure when using the first flush.

  5. Balanced sampling

    NARCIS (Netherlands)

    Brus, D.J.

    2015-01-01

    In balanced sampling a linear relation between the soil property of interest and one or more covariates with known means is exploited in selecting the sampling locations. Recent developments make this sampling design attractive for statistical soil surveys. This paper introduces balanced sampling

  6. Electron radiography

    Science.gov (United States)

    Merrill, Frank E.; Morris, Christopher

    2005-05-17

    A system capable of performing radiography using a beam of electrons. Diffuser means receive a beam of electrons and diffuse the electrons before they enter first matching quadrupoles where the diffused electrons are focused prior to the diffused electrons entering an object. First imaging quadrupoles receive the focused diffused electrons after the focused diffused electrons have been scattered by the object for focusing the scattered electrons. Collimator means receive the scattered electrons and remove scattered electrons that have scattered to large angles. Second imaging quadrupoles receive the collimated scattered electrons and refocus the collimated scattered electrons and map the focused collimated scattered electrons to transverse locations on an image plane representative of the electrons' positions in the object.

  7. Evaluation of BacLite Rapid MRSA, a rapid culture based screening test for the detection of ciprofloxacin and methicillin resistant S. aureus (MRSA from screening swabs

    Directory of Open Access Journals (Sweden)

    Skyrme Margaret

    2006-09-01

    Full Text Available Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA is a major nosocomial pathogen worldwide. The need for accurate and rapid screening methods to detect MRSA carriers has been clearly established. The performance of a novel assay, BacLite Rapid MRSA (Acolyte Biomedica, UK for the rapid detection (5 h and identification of hospital associated ciprofloxacin resistant strains of MRSA directly from nasal swab specimens was compared to that obtained by culture on Mannitol salt agar containing Oxacillin (MSAO after 48 h incubation. Results A total of 1382 nasal screening swabs were tested by multiple operators. The BacLite Rapid MRSA test detected 142 out of the 157 confirmed MRSA that were detected on MSAO giving a diagnostic sensitivity of 90.4, diagnostic specificity of 95.7% and a negative predictive value of 98.7%. Of the 15 false negatives obtained by the BacLite Rapid MRSA test, seven grew small amounts ( Conclusion The Baclite MRSA test is easy to use and provides a similar level of sensitivity to conventional culture for the detection of nasal carriage of MRSA with the advantage that the results are obtained much more rapidly.

  8. Comparison of Gas Chromatography-Mass Spectrometry and Gas Chromatography-Tandem Mass Spectrometry with Electron Ionization and Negative-Ion Chemical Ionization for Analyses of Pesticides at Trace Levels in Atmospheric Samples

    Directory of Open Access Journals (Sweden)

    Renata Raina

    2008-01-01

    Full Text Available A comparison of detection limits of gas chromatography-mass spectrometry (GC-MS in selected ion monitoring (SIM with gas chromatography-tandem mass spectrometry (GC-MS/MS in selected reaction monitoring (SRM mode with both electron ionization (EI and negative-ion chemical ionization (NCI are presented for over 50 pesticides ranging from organochlorines (OCs, organophosphorus pesticides (OPs and pre-emergent herbicides used in the Canadian prairies (triallate, trifluralin, ethalfluralin. The developed GC-EI/SIM, GC-NCI/SIM, and GC-NCI/SRM are suitable for the determination of pesticides in air sample extracts at concentrations <100 pg µL−1 (<100 pg m−3 in air. No one method could be used to analyze the range of pre-emergent herbicides, OPs, and OCs investigated. In general GC-NCI/SIM provided the lowest method detection limits (MDLs commonly 2.5–10 pg µL−1 along with best confirmation (<25% RSD of ion ratio, while GC-NCI/SRM is recommended for use where added selectivity or confirmation is required (such as parathion-ethyl, tokuthion, carbofenothion. GC-EI/SRM at concentration <100 pg µL−1 was not suitable for most pesticides. GC-EI/SIM was more prone to interference issues than NCI methods, but gave good sensitivity (MDLs 1–10 pg µL−1 for pesticides with poor NCI response (OPs: sulfotep, phorate, aspon, ethion, and OCs: alachlor, aldrin, perthane, and DDE, DDD, DDT.

  9. Fast electron microscopy via compressive sensing

    Science.gov (United States)

    Larson, Kurt W; Anderson, Hyrum S; Wheeler, Jason W

    2014-12-09

    Various technologies described herein pertain to compressive sensing electron microscopy. A compressive sensing electron microscope includes a multi-beam generator and a detector. The multi-beam generator emits a sequence of electron patterns over time. Each of the electron patterns can include a plurality of electron beams, where the plurality of electron beams is configured to impart a spatially varying electron density on a sample. Further, the spatially varying electron density varies between each of the electron patterns in the sequence. Moreover, the detector collects signals respectively corresponding to interactions between the sample and each of the electron patterns in the sequence.

  10. Efficient recovery of fluoroquinolone-susceptible and fluoroquinolone-resistant Escherichia coli strains from frozen samples.

    Science.gov (United States)

    Lautenbach, Ebbing; Santana, Evelyn; Lee, Abby; Tolomeo, Pam; Black, Nicole; Babson, Andrew; Perencevich, Eli N; Harris, Anthony D; Smith, Catherine A; Maslow, Joel

    2008-04-01

    We assessed the rate of recovery of fluoroquinolone-resistant and fluoroquinolone-susceptible Escherichia coli isolates from culture of frozen perirectal swab samples compared with the results for culture of the same specimen before freezing. Recovery rates for these 2 classes of E. coli were 91% and 83%, respectively. The majority of distinct strains recovered from the initial sample were also recovered from the frozen sample. The strains that were not recovered were typically present only in low numbers in the initial sample. These findings emphasize the utility of frozen surveillance samples.

  11. Evaluation of a novel real-time PCR test based on the ssrA gene for the identification of group B streptococci in vaginal swabs.

    LENUS (Irish Health Repository)

    Wernecke, Martina

    2009-01-01

    BACKGROUND: Despite the implementation of prevention guidelines, early-onset group B streptococci (GBS) disease remains a cause of neonatal morbidity and mortality worldwide. Strategies to identify women who are at risk of transmitting GBS to their infant and the administration of intrapartum antibiotics have greatly reduced the incidence of neonatal GBS disease. However, there is a requirement for a rapid diagnostic test for GBS that can be carried out in a labour ward setting especially for women whose GBS colonisation status is unknown at the time of delivery. We report the design and evaluation of a real-time PCR test (RiboSEQ GBS test) for the identification of GBS in vaginal swabs from pregnant women. METHODS: The qualitative real-time PCR RiboSEQ GBS test was designed based on the bacterial ssrA gene and incorporates a competitive internal standard control. The analytical sensitivity of the test was established using crude lysate extracted from serial dilutions of overnight GBS culture using the IDI Lysis kit. Specificity studies were performed using DNA prepared from a panel of GBS strains, related streptococci and other species found in the genital tract environment. The RiboSEQ GBS test was evaluated on 159 vaginal swabs from pregnant women and compared with the GeneOhm StrepB Assay and culture for the identification of GBS. RESULTS: The RiboSEQ GBS test is specific and has an analytical sensitivity of 1-10 cell equivalents. The RiboSEQ GBS test was 96.4% sensitive and 95.8% specific compared to "gold standard" culture for the identification of GBS in vaginal swabs from pregnant women. In this study, the RiboSEQ GBS test performed slightly better than the commercial BD GeneOhm StrepB Assay which gave a sensitivity of 94.6% and a specificity of 89.6% compared to culture. CONCLUSION: The RiboSEQ GBS test is a valuable method for the rapid, sensitive and specific detection of GBS in pregnant women. This study also validates the ssrA gene as a suitable and

  12. Language sampling

    DEFF Research Database (Denmark)

    Rijkhoff, Jan; Bakker, Dik

    1998-01-01

    This article has two aims: [1] to present a revised version of the sampling method that was originally proposed in 1993 by Rijkhoff, Bakker, Hengeveld and Kahrel, and [2] to discuss a number of other approaches to language sampling in the light of our own method. We will also demonstrate how our...... sampling method is used with different genetic classifications (Voegelin & Voegelin 1977, Ruhlen 1987, Grimes ed. 1997) and argue that —on the whole— our sampling technique compares favourably with other methods, especially in the case of exploratory research....

  13. Culture-independent genome sequencing of clinical samples reveals an unexpected heterogeneity of infections by Chlamydia pecorum.

    Science.gov (United States)

    Bachmann, Nathan L; Sullivan, Mitchell J; Jelocnik, Martina; Myers, Garry S A; Timms, Peter; Polkinghorne, Adam

    2015-05-01

    Chlamydia pecorum is an important global pathogen of livestock, and it is also a significant threat to the long-term survival of Australia's koala populations. This study employed a culture-independent DNA capture approach to sequence C. pecorum genomes directly from clinical swab samples collected from koalas with chlamydial disease as well as from sheep with arthritis and conjunctivitis. Investigations into single-nucleotide polymorphisms within each of the swab samples revealed that a portion of the reads in each sample belonged to separate C. pecorum strains, suggesting that all of the clinical samples analyzed contained mixed populations of genetically distinct C. pecorum isolates. This observation was independent of the anatomical site sampled and the host species. Using the genomes of strains identified in each of these samples, whole-genome phylogenetic analysis revealed that a clade containing a bovine and a koala isolate is distinct from other clades comprised of livestock or koala C. pecorum strains. Providing additional evidence to support exposure of koalas to Australian livestock strains, two minor strains assembled from the koala swab samples clustered with livestock strains rather than koala strains. Culture-independent probe-based genome capture and sequencing of clinical samples provides the strongest evidence yet to suggest that naturally occurring chlamydial infections are comprised of multiple genetically distinct strains. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  14. Culture-independent quantification of Salmonella enterica in carcass gauze swabs by flotation prior to real-time PCR

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Schelin, Jenny; Norling, Börje

    2011-01-01

    To facilitate quantitative risk assessment in the meat production chain, there is a need for culture-independent quantification methods. The aim of this study was to evaluate the use of flotation, a non-destructive sample preparation method based on traditional buoyant density centrifugation, for...

  15. The use of culture, pooled samples and PCR for identification of herds infected with Brachyspira hyodysenteriae.

    Science.gov (United States)

    Fellström, C; Zimmerman, U; Aspan, A; Gunnarsson, A

    2001-06-01

    The sensitivity of culturing Brachyspira hyodysenteriae was determined after sampling with swabs from porcine fecal specimens inoculated with tenfold dilutions of a field strain of these microbes. After storage of swabs, Brachyspira hyodysenteriae was recovered throughout the first 3 weeks after inoculation from feces with more than 140 cells/g. Viable spirochetes could still be recovered after up to 83 days of storage from feces, with 1.4 x 10(6) cells or more per gram. Culture for Brachyspira spp. was performed on 285 rectal swabs, which were pooled in batches of five. The number of pooled samples positive for B. hyodysenteriae corresponded with the sum results of individual analysis of the corresponding collections of five samples. A PCR system based on the tlyA gene of B. hyodysenteriae was developed and tested on primary cultures of pooled samples. The results of the PCR assay showed a 97% correlation with the culture results. The prevalence of Brachyspira spp. was determined in five swine herds and found to be highest among breeding gilts and boars aged 13-16 weeks and among 6-12-week-old weaned pigs. In contrast, Brachyspira spp. were only rarely found in sows, which may reflect the development of immunity by adult pigs to all species of the genus.

  16. Occurence and antimicrobial resistance of Arcobacter species in food and slaughterhouse samples

    Directory of Open Access Journals (Sweden)

    Mehmet ELMALI

    Full Text Available Abstract The objectives of this study were i to isolate Arcobacter species (Arcobacter butzleri, Arcobacter skirrowii, Arcobacter cryaerophilus from different foods and sources, ii to verify the isolates by multiplex PCR assay, iii to detect the antibiotic resistance profiles of the isolates. In this study a total of 60 Arcobacter isolates were obtained. Arcobacter species were mostly isolated from swab samples (40%, followed by wastewater (29.1%, broiler wing meat (30%, raw milk (23.9% and minced meat (6.6%. Regarding the seasonal distribution of Arcobacter from swab and wastewater samples, the bacterium was commonly isolated from wastewater in winter and spring, while it was frequently detected in swab samples during autumn and spring. All of the isolates were found to be resistant to nalidixic acid, ampicillin, rifampin, and erythromycin. The most effective antibiotic was tetracycline, because 96.66% of the isolates were susceptible against it. This is the first report of the isolation, seasonal distribution and antimicrobial susceptibility of Arcobacter species in cattle slaughterhouse samples in Turkey. These results indicate that foods of animal origin and cattle slaughterhouses are significant source of the antimicrobial resistant arcobacters.

  17. Venous Sampling

    Science.gov (United States)

    ... neck to help locate abnormally functioning glands or pituitary adenoma . This test is most often used after an unsuccessful neck exploration. Inferior petrosal sinus sampling , in which blood samples are taken from veins that drain the pituitary gland to study disorders related to pituitary hormone ...

  18. Sampling Development

    Science.gov (United States)

    Adolph, Karen E.; Robinson, Scott R.

    2011-01-01

    Research in developmental psychology requires sampling at different time points. Accurate depictions of developmental change provide a foundation for further empirical studies and theories about developmental mechanisms. However, overreliance on widely spaced sampling intervals in cross-sectional and longitudinal designs threatens the validity of…

  19. Language sampling

    DEFF Research Database (Denmark)

    Rijkhoff, Jan; Bakker, Dik

    1998-01-01

    This article has two aims: [1] to present a revised version of the sampling method that was originally proposed in 1993 by Rijkhoff, Bakker, Hengeveld and Kahrel, and [2] to discuss a number of other approaches to language sampling in the light of our own method. We will also demonstrate how our...

  20. Environmental sampling

    Energy Technology Data Exchange (ETDEWEB)

    Puckett, J.M.

    1998-12-31

    Environmental Sampling (ES) is a technology option that can have application in transparency in nuclear nonproliferation. The basic process is to take a sample from the environment, e.g., soil, water, vegetation, or dust and debris from a surface, and through very careful sample preparation and analysis, determine the types, elemental concentration, and isotopic composition of actinides in the sample. The sample is prepared and the analysis performed in a clean chemistry laboratory (CCL). This ES capability is part of the IAEA Strengthened Safeguards System. Such a Laboratory is planned to be built by JAERI at Tokai and will give Japan an intrinsic ES capability. This paper presents options for the use of ES as a transparency measure for nuclear nonproliferation.

  1. Self-taken vaginal swabs versus clinician-taken for detection of candida and bacterial vaginosis: a case-control study in primary care.

    Science.gov (United States)

    Barnes, Pam; Vieira, Rute; Harwood, Jayne; Chauhan, Mayur

    2017-12-01

    Vaginal discharge and vulvitis are common presenting symptoms in general practice. Few studies have specifically looked at the validity of self-taken low vulvovaginal swabs (LVS) for the diagnosis of vulvovaginal candidiasis (VVC) and bacterial vaginosis (BV). To assess if patient self-taken LVS are a valid alternative to clinician-taken high vaginal swabs (HVS) for the detection of VVC and BV. Case-control study with the patient acting as their own control in an urban sexual health centre in Newcastle upon Tyne, UK. Females aged 16-65 years attending with symptomatic vaginal discharge, vulval irritation, genital pain, and an offensive genital smell were recruited into the study. Participants took a self-taken LVS before vaginal examination, during which a clinician took an HVS (reference standard). Main outcome measures were the diagnosis of BV or VVC infection. A total of 104 females were enrolled. Of those, 45 were diagnosed with VVC and 26 with BV. The sensitivities of self-taken LVS for VVC and BV were 95.5% and 88.5% respectively. Cohen's κ coefficient showed 'strong agreement' for the detection of both VVC and BV. Vulval itching was the most common symptom associated with VVC (69%), whereas 50% of females diagnosed with BV presented with an offensive discharge. Both symptoms had poor positive predictive values (0.63 and 0.50, respectively). Self-taken LVS appears to be a valid alternative to clinician-taken HVS for detecting VVC and BV infections. Symptoms were found to be a poor indicator of underlying infection. © British Journal of General Practice 2017.

  2. LC-MS method development and comparison of sampling materials for the analysis of organic gunshot residues.

    Science.gov (United States)

    Gassner, Anne-Laure; Weyermann, Céline

    2016-07-01

    This study aimed at developing a LC-MS method to compare the efficiency of various sampling materials for the collection and subsequent analysis of organic gunshot residues (OGSR). Seven sampling materials, namely two "swab"-type and five "stub"-type collection materials, were tested. The evaluation of sampling materials was systematically carried out by first analyzing blank extracts of the materials to check for potential interferences and determining matrix effects. Based on these results, the best four materials, namely cotton buds, polyester swabs, a tape from 3M and PTFE were compared in terms of collection efficiency during shooting experiments using a set of 9mm Luger ammunition. It was found that the tape was capable of recovering the highest amounts of OGSR. As tape-lifting is the technique currently used in routine for inorganic GSR, OGSR analysis might be implemented without modifying IGSR sampling and analysis procedure. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  3. Novel real-time PCR assays using TaqMan minor groove binder probes for identification of fecal carriage of Streptococcus bovis/Streptococcus equinus complex from rectal swab specimens.

    Science.gov (United States)

    Lopes, Paulo Guilherme Markus; Cantarelli, Vlademir Vicente; Agnes, Grasiela; Costabeber, Ane Micheli; d'Azevedo, Pedro Alves

    2014-03-01

    Real-time PCR based on the recN and gyrB genes was developed to detect four Streptococcus bovis/Streptococcus equinus complex (SBEC) subspecies from rectal swab specimens. The overall prevalence was 35.2%: Streptococcus gallolyticus subsp. gallolyticus (11.1%), S. gallolyticus subsp. pasteurianus (13%), Streptococcus infantarius subsp. coli (20.4%), and S. infantarius subsp. infantarius (11.1%). To conclude, these real-time PCR assays provide a reliable molecular method to detect SBEC pathogenic subspecies from rectal swab specimens.

  4. Effectiveness of hand hygiene education among a random sample of women from the community

    OpenAIRE

    Ubheeram, J.; Biranjia-Hurdoyal, S.D.

    2017-01-01

    Summary Objective. The effectiveness of hand hygiene education was investigated by studying the hand hygiene awareness and bacterial hand contamination among a random sample of 170 women in the community. Methods. Questionnaire was used to assess the hand hygiene awareness score, followed by swabbing of the dominant hand. Bacterial identification was done by conventional biochemical tests. Results. Better hand hygiene awareness score was significantly associated with age, scarce bacterial gro...

  5. Techniques for the detection of pathogenic Cryptococcus species in wood decay substrata and the evaluation of viability in stored samples

    Directory of Open Access Journals (Sweden)

    Christian Alvarez

    2013-02-01

    Full Text Available In this study, we evaluated several techniques for the detection of the yeast form of Cryptococcus in decaying wood and measured the viability of these fungi in environmental samples stored in the laboratory. Samples were collected from a tree known to be positive for Cryptococcus and were each inoculated on 10 Niger seed agar (NSA plates. The conventional technique (CT yielded a greater number of positive samples and indicated a higher fungal density [in colony forming units per gram of wood (CFU.g-1] compared to the humid swab technique (ST. However, the difference in positive and false negative results between the CT-ST was not significant. The threshold of detection for the CT was 0.05.10³ CFU.g-1, while the threshold for the ST was greater than 0.1.10³ CFU-1. No colonies were recovered using the dry swab technique. We also determined the viability of Cryptococcus in wood samples stored for 45 days at 25ºC using the CT and ST and found that samples not only continued to yield a positive response, but also exhibited an increase in CFU.g-1, suggesting that Cryptococcus is able to grow in stored environmental samples. The ST.1, in which samples collected with swabs were immediately plated on NSA medium, was more efficient and less laborious than either the CT or ST and required approximately 10 min to perform; however, additional studies are needed to validate this technique.

  6. Elevating sampling

    Science.gov (United States)

    Labuz, Joseph M.; Takayama, Shuichi

    2014-01-01

    Sampling – the process of collecting, preparing, and introducing an appropriate volume element (voxel) into a system – is often under appreciated and pushed behind the scenes in lab-on-a-chip research. What often stands in the way between proof-of-principle demonstrations of potentially exciting technology and its broader dissemination and actual use, however, is the effectiveness of sample collection and preparation. The power of micro- and nanofluidics to improve reactions, sensing, separation, and cell culture cannot be accessed if sampling is not equally efficient and reliable. This perspective will highlight recent successes as well as assess current challenges and opportunities in this area. PMID:24781100

  7. Self-sampling for human papillomavirus testing among rural young women of KwaZulu-Natal, South Africa.

    Science.gov (United States)

    Mbatha, J N; Galapaththi-Arachchige, H N; Mtshali, A; Taylor, M; Ndhlovu, P D; Kjetland, E F; Baay, M F D; Mkhize-Kwitshana, Z L

    2017-12-06

    Cervical cancer is a major problem in women and it is important to find a suitable and acceptable screening method, especially among young in low-resource areas for future human papillomavirus (HPV) vaccine follow-up investigations. The study sought to test the acceptability of self-sampling as well as the suitability of the specimen collecting devices. Ninety-eight young women from rural KwaZulu-Natal were enrolled between March and July 2014. Collected genital specimens were transferred to colour indicator cards for HPV detection. Participants answered a questionnaire where they described their experiences with self-sampling. Samples were tested for high-risk HPV using GP5/6+ PCR. Of the enrolled participants, 91 answered questionnaires and indicated that self-sampling was preferred by 51/91 (56%) women while 40/91 (44%) indicated preference for sampling by a doctor (p = 0.023). The majority, 64% were comfortable using a swab, 22% preferred a brush while 11% were comfortable with both devices. Of the 98 self-sampled specimens 61 were negative for HPV in both specimens while 37 were HPV-positive in either brush or swab. Of the 37, 26 (70%) were HPV-positive in both brush and swab (kappa = 0.743) and 11 (30%) were discordant. Self-sampling was acceptable to the majority of participants in this rural area. The Dacron swab was the preferred device, and can be used in combination with colour indicator cards for comfortable self-sampling, easy storage and transport of specimens plus detection.

  8. New methods for electron tomography

    NARCIS (Netherlands)

    Ziese, Ulrike

    2002-01-01

    Electron tomography is a method for obtaining three-dimensional structural information from electron micrographs. It can be applied to a wide range of samples that can be prepared for transmission electron microscopy (TEM)—may they be of biological origin like e.g. cryo or thin plastic sections of

  9. Identification of chemical warfare agents from vapor samples using a field-portable capillary gas chromatography/membrane-interfaced electron ionization quadrupole mass spectrometry instrument with Tri-Bed concentrator.

    Science.gov (United States)

    Nagashima, Hisayuki; Kondo, Tomohide; Nagoya, Tomoki; Ikeda, Toru; Kurimata, Naoko; Unoke, Shohei; Seto, Yasuo

    2015-08-07

    A field-portable gas chromatograph-mass spectrometer (Hapsite ER system) was evaluated for the detection of chemical warfare agents (CWAs) in the vapor phase. The system consisted of Tri-Bed concentrator gas sampler (trapping time: 3s(-1)min), a nonpolar low thermal-mass capillary gas chromatography column capable of raising temperatures up to 200°C, a hydrophobic membrane-interfaced electron ionization quadrupole mass spectrometer evacuated by a non-evaporative getter pump for data acquisition, and a personal computer for data analysis. Sample vapors containing as little as 22μg sarin (GB), 100μg soman (GD), 210μg tabun (GA), 55μg cyclohexylsarin (GF), 4.8μg sulfur mustard, 390μg nitrogen mustard 1, 140μg of nitrogen mustard 2, 130μg nitrogen mustard 3, 120μg of 2-chloroacetophenone and 990μg of chloropicrin per cubic meter could be confirmed after Tri-Bed micro-concentration (for 1min) and automated AMDIS search within 12min. Using manual deconvolution by background subtraction of neighboring regions on the extracted ion chromatograms, the above-mentioned CWAs could be confirmed at lower concentration levels. The memory effects were also examined and we found that blister agents showed significantly more carry-over than nerve agents. Gasoline vapor was found to interfere with the detection of GB and GD, raising the concentration limits for confirmation in the presence of gasoline by both AMDIS search and manual deconvolution; however, GA and GF were not subject to interference by gasoline. Lewisite 1, and o-chlorobenzylidene malononitrile could also be confirmed by gas chromatography, but it was hard to quantify them. Vapors of phosgene, chlorine, and cyanogen chloride could be confirmed by direct mass spectrometric detection at concentration levels higher than 2, 140, and 10mg/m(3) respectively, by bypassing the micro-concentration trap and gas chromatographic separation. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Electron Microscopy.

    Science.gov (United States)

    Beer, Michael

    1980-01-01

    Reviews technical aspects of structure determination in biological electron microscopy (EM). Discusses low dose EM, low temperature microscopy, electron energy loss spectra, determination of mass or molecular weight, and EM of labeled systems. Cites 34 references. (CS)

  11. Electronic Commerce

    OpenAIRE

    Slavko Đerić

    2016-01-01

    Electronic commerce can be defined in different ways. Any definition helps to understand and explain that concept as better as possible.. Electronic commerce is a set of procedures and technologies that automate the tasks of financial transactions using electronic means. Also, according to some authors, electronic commerce is defined as a new concept, which is being developed and which includes process of buying and selling or exchanging products, services or information via computer networks...

  12. Understand electronics

    CERN Document Server

    Bishop, Owen

    2013-01-01

    Understand Electronics provides a readable introduction to the exciting world of electronics for the student or enthusiast with little previous knowledge. The subject is treated with the minimum of mathematics and the book is extensively illustrated.This is an essential guide for the newcomer to electronics, and replaces the author's best-selling Beginner's Guide to Electronics.The step-by-step approach makes this book ideal for introductory courses such as the Intermediate GNVQ.

  13. Vacuum electronics

    CERN Document Server

    Eichmeier, Joseph A

    2008-01-01

    Nineteen experts from the electronics industry, research institutes and universities have joined forces to prepare this book. ""Vacuum Electronics"" covers the electrophysical fundamentals, the present state of the art and applications, as well as the future prospects of microwave tubes and systems, optoelectronics vacuum devices, electron and ion beam devices, light and X-ray emitters, particle accelerators and vacuum interrupters. These topics are supplemented by useful information about the materials and technologies of vacuum electronics and vacuum technology.

  14. Electronic components

    CERN Document Server

    Colwell, Morris A

    1976-01-01

    Electronic Components provides a basic grounding in the practical aspects of using and selecting electronics components. The book describes the basic requirements needed to start practical work on electronic equipment, resistors and potentiometers, capacitance, and inductors and transformers. The text discusses semiconductor devices such as diodes, thyristors and triacs, transistors and heat sinks, logic and linear integrated circuits (I.C.s) and electromechanical devices. Common abbreviations applied to components are provided. Constructors and electronics engineers will find the book useful

  15. ELECTRONIC SIGNATURES

    African Journals Online (AJOL)

    10332324

    'electronic signature' means data attached to, incorporated in, or logically associated with other data and which is intended by the user to serve as a signature;. The suggested new definition for an electronic signature reads as follows: 'electronic signature' means a sound, symbol or process that is (i) uniquely linked to the ...

  16. ELECTRONIC SIGNATURES

    African Journals Online (AJOL)

    10332324

    (a) facilitate ecommerce;2. (b) remove and prevent barriers to electronic communications in South Africa;3. (c) ensure that electronic transactions in the Republic conform to the highest international standards;4. (d) promote the development of electronic transactions services which are responsive to the needs of users and ...

  17. Development and evaluation of the quantitative real-time PCR assay in detection and typing of herpes simplex virus in swab specimens from patients with genital herpes.

    Science.gov (United States)

    Liu, Junlian; Yi, Yong; Chen, Wei; Si, Shaoyan; Yin, Mengmeng; Jin, Hua; Liu, Jianjun; Zhou, Jinlian; Zhang, Jianzhong

    2015-01-01

    Genital herpes (GH), which is caused mainly by herpes simplex virus (HSV)-2 and HSV-1, remains a worldwide problem. Laboratory confirmation of GH is important, particularly as there are other conditions which present similarly to GH, while atypical presentations of GH also occur. Currently, virus culture is the classical method for diagnosis of GH, but it is time consuming and with low sensitivity. A major advance for diagnosis of GH is to use Real-time polymerase chain reaction (PCR). In this study, to evaluate the significance of the real-time PCR method in diagnosis and typing of genital HSV, the primers and probes targeted at HSV-1 DNA polymerase gene and HSV-2 glycoprotein D gene fraction were designed and applied to amplify DNA from HSV-1 or HSV-2 by employing the real-time PCR technique. Then the PCR reaction system was optimized and evaluated. HSV in swab specimens from patients with genital herpes was detected by real-time PCR. The real-time PCR assay showed good specificity for detection and typing of HSV, with good linear range (5×10(2)~5×10(8) copies/ml, r=0.997), a sensitivity of 5×10(2) copies/ml, and good reproducibility (intra-assay coefficients of variation 2.29% and inter-assay coefficients of variation 4.76%). 186 swab specimens were tested for HSV by real-time PCR, and the positive rate was 23.7% (44/186). Among the 44 positive specimens, 8 (18.2%) were positive for HSV-1 with a viral load of 8.5546×10(6) copies/ml and 36 (81.2%) were positive for HSV-2 with a viral load of 1.9861×10(6) copies/ml. It is concluded that the real-time PCR is a specific, sensitive and rapid method for the detection and typing of HSV, which can be widely used in clinical diagnosis of GH.

  18. Nanopipettes: probes for local sample analysis† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c5sc00668f Click here for additional data file.

    Science.gov (United States)

    Saha-Shah, Anumita; Weber, Anna E.; Karty, Jonathan A.; Ray, Steven J.; Hieftje, Gary M.

    2015-01-01

    Nanopipettes (pipettes with diameters nanopipette shank was studied to optimize sampling volume and probe geometry. This method was utilized to collect nanoliter volumes (nanopipettes for surface sampling of mouse brain tissue sections was also explored. Lipid analyses were performed on mouse brain tissues with spatial resolution of sampling as small as 50 μm. Nanopipettes were shown to be a versatile tool that will find further application in studies of sample heterogeneity and population analysis for a wide range of samples. PMID:28706697

  19. Sticker electronics

    KAUST Repository

    Hussain, Muhammad Mustafa

    2017-09-08

    Electronic stickers may be manufactured on flexible substrates (110, 120, 130) as layers and packaged together. The package may then have an adhesive applied to one side to provide capability for sticking the electronic devices to surfaces. The stickers can be wrappable, placed on surfaces, glued on walls or mirrors or wood or stone, and have electronics (112, 122, 132) which may or may not be ultrathin. Packaging for the electronic sticker can use polymer on cellulose manufacturing and/or three dimensional (3-D) printing. The electronic stickers may provide lighting capability, sensing capability, and/or recharging capabilities.

  20. Basic electronics

    CERN Document Server

    Holbrook, Harold D

    1971-01-01

    Basic Electronics is an elementary text designed for basic instruction in electricity and electronics. It gives emphasis on electronic emission and the vacuum tube and shows transistor circuits in parallel with electron tube circuits. This book also demonstrates how the transistor merely replaces the tube, with proper change of circuit constants as required. Many problems are presented at the end of each chapter. This book is comprised of 17 chapters and opens with an overview of electron theory, followed by a discussion on resistance, inductance, and capacitance, along with their effects on t

  1. Electronic Government and Electronic Participation

    NARCIS (Netherlands)

    Tambouris, E.; Scholl, H.J.; Janssen, M.F.W.H.A.; Wimmer, M.A.; Tarabanis, K.; Gascó, M.; Klievink, A.J.; Lindgren, I.; Milano, M.; Panagiotopoulos, P.; Pardo, T.A.; Parycek, P.; Sæbø, O.

    2015-01-01

    Electronic government and electronic participation continue to transform the public sector and society worldwide and are constantly being transformed themselves by emerging information and communication technologies. This book presents papers from the 14th International Federation for Information

  2. Electronic Government and Electronic Participation

    NARCIS (Netherlands)

    Tambouris, E; Scholl, H.J.; Janssen, M.F.W.H.A.; Wimmer, M.A.; Tarabanis, K; Gascó, M; Klievink, A.J.; Lindgren, I; Milano, M; Panagiotopoulos, P; Pardo, T.A.; Parycek, P; Sæbø, Ø

    2016-01-01

    Electronic government and electronic participation continue to transform the public sector and society worldwide and are constantly being transformed themselves by emerging information and communication technologies.This book presents papers from the 14th International Federation for Information

  3. Electron Tree

    DEFF Research Database (Denmark)

    Appelt, Ane L; Rønde, Heidi S

    2013-01-01

    The photo shows a close-up of a Lichtenberg figure – popularly called an “electron tree” – produced in a cylinder of polymethyl methacrylate (PMMA). Electron trees are created by irradiating a suitable insulating material, in this case PMMA, with an intense high energy electron beam. Upon discharge......, during dielectric breakdown in the material, the electrons generate branching chains of fractures on leaving the PMMA, producing the tree pattern seen. To be able to create electron trees with a clinical linear accelerator, one needs to access the primary electron beam used for photon treatments. We...... appropriated a linac that was being decommissioned in our department and dismantled the head to circumvent the target and ion chambers. This is one of 24 electron trees produced before we had to stop the fun and allow the rest of the accelerator to be disassembled....

  4. [Atypical agents of wound infection and targeted samples].

    Science.gov (United States)

    Kucisec-Tepes, Nastja

    2012-10-01

    necessary to be thorough, meticulous, and conduct revision of the patient, diagnosis, treatment and samples. Crucial for the detection of the agents atypical wound infection is discussion and agreement with clinical microbiologist. Essential for the quality diagnosis is decontamination of the wound before taking targeted samples. The targeted sample is biopsy specimen as the "gold standard", less frequently aspirate, depending on the quantity and content of the wound. Swab as a sample is not recommended. If there is no other choice, only exceptionally surface swabs may be taken, but only under the following conditions: decontamination of the wound with the application of Levine's or Z-technique of taking of swabs.

  5. Comparison of the BD MAX MRSA XT to the Cepheid™ Xpert® MRSA assay for the molecular detection of methicillin-resistant Staphylococcus aureus from nasal swabs.

    Science.gov (United States)

    Mehta, Sanjay R; Estrada, Jasmine; Ybarra, Juan; Fierer, Joshua

    2017-04-01

    Variation in MRSA genotypes may affect the sensitivity of molecular assays to detect this organism. We compared 2 commonly used screening assays, the Cepheid™ Xpert® MRSA and the BD MAX™ MRSA XT on consecutively obtained nasal swabs from 479 subjects. Specimens giving discordant results were subjected to additional microbiologic and molecular testing. Six hundred forty-two (97.6%) of the 658 test results were concordant. Of the 16 discordant results from 12 subjects, additional results suggested that 9 (60%) of the 15 MRSA XT assays were likely correct, and 6 (40%) of the 15 Xpert® assays were likely correct. One discordant result could not be resolved. A mecA dropout and novel mec right-extremity junction (MREJ) sites led to false-positive and negative results by Xpert®. While both assays performed well, continued vigilance is needed to monitor for Staphylococcus aureus with novel MREJ sites, mecA dropouts, and mecC, leading to inaccurate results in screening assays. Published by Elsevier Inc.

  6. Hand Hygiene Practices and Microbial Investigation of Hand Contact Swab among Physiotherapists in an Ebola Endemic Region: Implications for Public Health

    Science.gov (United States)

    Maduako, V.; Ibeneme, G. C.; Ezuma, A.; Ettu, T. U.; Onyemelukwe, N. F.; Fortwengel, G.

    2017-01-01

    Background Hand hygiene practices (HHP), as a critical component of infection prevention/control, were investigated among physiotherapists in an Ebola endemic region. Method A standardized instrument was administered to 44 randomly selected physiotherapists (23 males and 21 females), from three tertiary hospitals in Enugu, Nigeria. Fifteen participants (aged 22–59 years) participated in focus group discussions (FGDs) and comprised 19 participants in a subsequent laboratory study. After treatment, the palms/fingers of physiotherapists were swabbed and cultured, then incubated aerobically overnight at 37°C, and examined for microbial growths. An antibiogram of the bacterial isolates was obtained. Results The majority (34/77.3%) of physiotherapists were aware of the HHP protocol, yet only 15/44.1% rated self-compliance at 71–100%. FGDs identified forgetfulness/inadequate HHP materials/infrastructure as the major barriers to HHP. Staphylococcus aureus were the most prevalent organisms, prior to (8/53.33%) and after (4/26.67%) HPP, while Pseudomonas spp. were acquired thereafter. E. coli were the most antibiotic resistant microbes but were completely removed after HHP. Ciprofloxacin and streptomycin were the most effective antibiotics. Conclusion Poor implementation of HPP was observed due to inadequate materials/infrastructure/poor behavioral orientation. Possibly, some HPP materials were contaminated; hence, new microbes were acquired. Since HPP removed the most antibiotic resistant microbes, it might be more effective in infection control than antibiotic medication. PMID:28691027

  7. Hand Hygiene Practices and Microbial Investigation of Hand Contact Swab among Physiotherapists in an Ebola Endemic Region: Implications for Public Health

    Directory of Open Access Journals (Sweden)

    S. Ibeneme

    2017-01-01

    Full Text Available Background. Hand hygiene practices (HHP, as a critical component of infection prevention/control, were investigated among physiotherapists in an Ebola endemic region. Method. A standardized instrument was administered to 44 randomly selected physiotherapists (23 males and 21 females, from three tertiary hospitals in Enugu, Nigeria. Fifteen participants (aged 22–59 years participated in focus group discussions (FGDs and comprised 19 participants in a subsequent laboratory study. After treatment, the palms/fingers of physiotherapists were swabbed and cultured, then incubated aerobically overnight at 37°C, and examined for microbial growths. An antibiogram of the bacterial isolates was obtained. Results. The majority (34/77.3% of physiotherapists were aware of the HHP protocol, yet only 15/44.1% rated self-compliance at 71–100%. FGDs identified forgetfulness/inadequate HHP materials/infrastructure as the major barriers to HHP. Staphylococcus aureus were the most prevalent organisms, prior to (8/53.33% and after (4/26.67% HPP, while Pseudomonas spp. were acquired thereafter. E. coli were the most antibiotic resistant microbes but were completely removed after HHP. Ciprofloxacin and streptomycin were the most effective antibiotics. Conclusion. Poor implementation of HPP was observed due to inadequate materials/infrastructure/poor behavioral orientation. Possibly, some HPP materials were contaminated; hence, new microbes were acquired. Since HPP removed the most antibiotic resistant microbes, it might be more effective in infection control than antibiotic medication.

  8. Comparison of Antibiotic Resistance Profile between Salmonella Spp., Salmonella Enterica Ser. Typhimurium and Enteritidis and Escherichia Coli Isolated from Rectal Swabs of Chicken

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2011-05-01

    Full Text Available The aim of this experiment was comparing of antibiotic resistance profile between Salmonella spp. and Escherichia coli isolated from rectal swabs of chicken from conventional breeding. For the antibiotic susceptibility testing disk diffusion method was used. The both tested bacteria were exposed against thirteen antibiotics: ampicillin, piperacillin, cefotaxime, ceftriaxone, doripenem, meropenem, levofloxacin, ofloxacin, amikacin, gentamycin, tygecycline, tetracycline and chloramphenicol. For the identification of these strains, we used Chromogenic coliform agar, Triple sugar iron agar and biochemical test (ENTEROtest 24. We identified Salmonella spp. by used MicroSEQ® Salmonella spp. Detection Kit for identification of this strain in Step ONE Real Time PCR. In this study, we determined that Salmonella spp. was more resistant like Escherichia coli. The highest resistance had isolates of Salmonella spp. to levofloxacin (100% and to ofloxacin (100%. Also to ampicillin was resistance in Salmonella spp. isolates about 83%. Only in case of piperacillin was resistance in Salmonella spp. isolates lower (50% like in Escherichia coli isolates (66.6%. The both strains were 100 % sensitive to doripenem, meropenem, amikacin, gentamycin and tygecycline. Antibiotic resistance is a biological danger. Bacteria, which we study, are considered to reservoirs of resistant genes and they are facultative and obligate pathogens. If these pathogen bacteria cause diseases those these diseases are difficult to treat.

  9. Performance of the cobas MRSA/SA Test for Simultaneous Detection of Methicillin-Susceptible and Methicillin-Resistant Staphylococcus aureus From Nasal Swabs.

    Science.gov (United States)

    Peterson, Lance R; Woods, Christopher W; Davis, Thomas