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Sample records for droplet-in-oil pcr array

  1. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman, Johan Frederik; Pierik, A.

    2012-01-01

    Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the

  2. Comprehensive GMO detection using real-time PCR array: single-laboratory validation.

    Science.gov (United States)

    Mano, Junichi; Harada, Mioko; Takabatake, Reona; Furui, Satoshi; Kitta, Kazumi; Nakamura, Kosuke; Akiyama, Hiroshi; Teshima, Reiko; Noritake, Hiromichi; Hatano, Shuko; Futo, Satoshi; Minegishi, Yasutaka; Iizuka, Tayoshi

    2012-01-01

    We have developed a real-time PCR array method to comprehensively detect genetically modified (GM) organisms. In the method, genomic DNA extracted from an agricultural product is analyzed using various qualitative real-time PCR assays on a 96-well PCR plate, targeting for individual GM events, recombinant DNA (r-DNA) segments, taxon-specific DNAs, and donor organisms of the respective r-DNAs. In this article, we report the single-laboratory validation of both DNA extraction methods and component PCR assays constituting the real-time PCR array. We selected some DNA extraction methods for specified plant matrixes, i.e., maize flour, soybean flour, and ground canola seeds, then evaluated the DNA quantity, DNA fragmentation, and PCR inhibition of the resultant DNA extracts. For the component PCR assays, we evaluated the specificity and LOD. All DNA extraction methods and component PCR assays satisfied the criteria set on the basis of previous reports.

  3. Rice1265 Array: PCR check - RMOS | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us RMOS Rice1265 Array: PCR check Data detail Data name Rice1265 Array: PCR check DOI 10.18908/...lsdba.nbdc00194-004 Description of data contents List of the PCR check results about clones used in Rice1265... Microarray (Full insert version) / (3'UTR version) Data file File name: rmos_1265pcrcheck.csv.zip File URL:... ftp://ftp.biosciencedbc.jp/archive/rmos/LATEST/rmos_1265pcrcheck.csv.zip File si...ze: 11.7 KB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/rmos_1265pcrcheck#en Data acquisiti

  4. High throughput gene expression measurement with real time PCR in a microfluidic dynamic array.

    Directory of Open Access Journals (Sweden)

    Sandra L Spurgeon

    Full Text Available We describe a high throughput gene expression platform based on microfluidic dynamic arrays. This system allows 2,304 simultaneous real time PCR gene expression measurements in a single chip, while requiring less pipetting than is required to set up a 96 well plate. We show that one can measure the expression of 45 different genes in 18 tissues with replicates in a single chip. The data have excellent concordance with conventional real time PCR and the microfluidic dynamic arrays show better reproducibility than commercial DNA microarrays.

  5. The potential of TaqMan Array Cards for detection of multiple biological agents by real-time PCR.

    Directory of Open Access Journals (Sweden)

    Phillip A Rachwal

    Full Text Available The TaqMan Array Card architecture, normally used for gene expression studies, was evaluated for its potential to detect multiple bacterial agents by real-time PCR. Ten PCR assays targeting five biological agents (Bacillus anthracis, Burkholderia mallei, Burkholderia pseudomallei, Francisella tularensis, and Yersinia pestis were incorporated onto Array Cards. A comparison of PCR performance of each PCR in Array Card and singleplex format was conducted using DNA extracted from pure bacterial cultures. When 100 fg of agent DNA was added to Array Card channels the following levels of agent detection (where at least one agent PCR replicate returned a positive result were observed: Y. pestis 100%, B. mallei & F. tularensis 93%; B. anthracis 71%; B. pseudomallei 43%. For B. mallei & pseudomallei detection the BPM2 PCR, which detects both species, outperformed PCR assays specific to each organism indicating identification of the respective species would not be reproducible at the 100 fg level. Near 100% levels of detection were observed when 100 fg of DNA was added to each PCR in singleplex format with singleplex PCRs also returning sporadic positives at the 10 fg per PCR level. Before evaluating the use of Array Cards for the testing of environmental and clinical sample types, with potential levels of background DNA and PCR inhibitors, users would therefore have to accept a 10-fold reduction in sensitivity of PCR assays on the Array Card format, in order to benefit for the capacity to test multiple samples for multiple agents. A two PCR per agent strategy would allow the testing of 7 samples for the presence of 11 biological agents or 3 samples for 23 biological agents per card (with negative control channels.

  6. Centrifugal micro-channel array droplet generation for highly parallel digital PCR.

    Science.gov (United States)

    Chen, Zitian; Liao, Peiyu; Zhang, Fangli; Jiang, Mengcheng; Zhu, Yusen; Huang, Yanyi

    2017-01-17

    Stable water-in-oil emulsion is essential to digital PCR and many other bioanalytical reactions that employ droplets as microreactors. We developed a novel technology to produce monodisperse emulsion droplets with high efficiency and high throughput using a bench-top centrifuge. Upon centrifugal spinning, the continuous aqueous phase is dispersed into monodisperse droplet jets in air through a micro-channel array (MiCA) and then submerged into oil as a stable emulsion. We performed dPCR reactions with a high dynamic range through the MiCA approach, and demonstrated that this cost-effective method not only eliminates the usage of complex microfluidic devices and control systems, but also greatly suppresses the loss of materials and cross-contamination. MiCA-enabled highly parallel emulsion generation combines both easiness and robustness of picoliter droplet production, and breaks the technical challenges by using conventional lab equipment and supplies.

  7. Pathway-focused PCR array profiling of enriched populations of laser capture microdissected hippocampal cells after traumatic brain injury.

    Directory of Open Access Journals (Sweden)

    Deborah R Boone

    Full Text Available Cognitive deficits in survivors of traumatic brain injury (TBI are associated with irreversible neurodegeneration in brain regions such as the hippocampus. Comparative gene expression analysis of dying and surviving neurons could provide insight into potential therapeutic targets. We used two pathway-specific PCR arrays (RT2 Profiler Apoptosis and Neurotrophins & Receptors PCR arrays to identify and validate TBI-induced gene expression in dying (Fluoro-Jade-positive or surviving (Fluoro-Jade-negative pyramidal neurons obtained by laser capture microdissection (LCM. In the Apoptosis PCR array, dying neurons showed significant increases in expression of genes associated with cell death, inflammation, and endoplasmic reticulum (ER stress compared with adjacent, surviving neurons. Pro-survival genes with pleiotropic functions were also significantly increased in dying neurons compared to surviving neurons, suggesting that even irreversibly injured neurons are able to mount a protective response. In the Neurotrophins & Receptors PCR array, which consists of genes that are normally expected to be expressed in both groups of hippocampal neurons, only a few genes were expressed at significantly different levels between dying and surviving neurons. Immunohistochemical analysis of selected, differentially expressed proteins supported the gene expression data. This is the first demonstration of pathway-focused PCR array profiling of identified populations of dying and surviving neurons in the brain after TBI. Combining precise laser microdissection of identifiable cells with pathway-focused PCR array analysis is a practical, low-cost alternative to microarrays that provided insight into neuroprotective signals that could be therapeutically targeted to ameliorate TBI-induced neurodegeneration.

  8. 1-Million droplet array with wide-field fluorescence imaging for digital PCR.

    Science.gov (United States)

    Hatch, Andrew C; Fisher, Jeffrey S; Tovar, Armando R; Hsieh, Albert T; Lin, Robert; Pentoney, Stephen L; Yang, David L; Lee, Abraham P

    2011-11-21

    Digital droplet reactors are useful as chemical and biological containers to discretize reagents into picolitre or nanolitre volumes for analysis of single cells, organisms, or molecules. However, most DNA based assays require processing of samples on the order of tens of microlitres and contain as few as one to as many as millions of fragments to be detected. Presented in this work is a droplet microfluidic platform and fluorescence imaging setup designed to better meet the needs of the high-throughput and high-dynamic-range by integrating multiple high-throughput droplet processing schemes on the chip. The design is capable of generating over 1-million, monodisperse, 50 picolitre droplets in 2-7 minutes that then self-assemble into high density 3-dimensional sphere packing configurations in a large viewing chamber for visualization and analysis. This device then undergoes on-chip polymerase chain reaction (PCR) amplification and fluorescence detection to digitally quantify the sample's nucleic acid contents. Wide-field fluorescence images are captured using a low cost 21-megapixel digital camera and macro-lens with an 8-12 cm(2) field-of-view at 1× to 0.85× magnification, respectively. We demonstrate both end-point and real-time imaging ability to perform on-chip quantitative digital PCR analysis of the entire droplet array. Compared to previous work, this highly integrated design yields a 100-fold increase in the number of on-chip digitized reactors with simultaneous fluorescence imaging for digital PCR based assays.

  9. Development of real-time PCR array for simultaneous detection of eight human blood-borne viral pathogens.

    Directory of Open Access Journals (Sweden)

    Natalia Pripuzova

    Full Text Available BACKGROUND: Real-time PCR array for rapid detection of multiple viral pathogens should be highly useful in cases where the sample volume and the time of testing are limited, i.e. in the eligibility testing of tissue and organ donors. FINDINGS: We developed a real-time PCR array capable of simultaneously detecting eight human viral pathogens: human immunodeficiency virus types 1 and 2 (HIV-1 and -2, hepatitis B virus (HBV, hepatitis C virus (HCV, human T-cell leukemia virus-1 and -2 (HTLV-1 and -2, vaccinia virus (VACV and West Nile virus (WNV. One hundred twenty (120 primers were designed using a combination of bioinformatics approaches, and, after experimental testing, 24 primer sets targeting eight viral pathogens were selected to set up the array with SYBR Green chemistry. The specificity and sensitivity of the virus-specific primer sets selected for the array were evaluated using analytical panels with known amounts of viruses spiked into human plasma. The array detected: 10 genome equivalents (geq/ml of HIV-2 and HCV, 50 geq of HIV-1 (subtype B, HBV (genotype A and WNV. It detected 100-1,000 geq/ml of plasma of HIV-1 subtypes (A - G, group N and CRF (AE and AG isolates. Further evaluation with a panel consisting of 28 HIV-1 and HIV-2 clinical isolates revealed no cross-reactivity of HIV-1 or HIV-2 specific primers with another type of HIV. All 28 viral isolates were identified with specific primer sets targeting the most conserved genome areas. The PCR array correctly identified viral infections in a panel of 17 previously quantified clinical plasma samples positive for HIV-1, HCV or HBV at as low as several geq per PCR reaction. CONCLUSIONS: The viral array described here demonstrated adequate performance in the testing of donors' clinical samples. Further improvement in its sensitivity for the broad spectrum of HIV-1 subtypes is under development.

  10. Gut Low Density Array (GULDA), a novel qPCR approach to the study of the intestinal microbial ecosystem

    DEFF Research Database (Denmark)

    Bergström, Anders; Andersen, Jens Bo; Licht, Tine Rask

    developed GULDA; a qPCR low-density array with particular focus on bacteria of relevance to the human gut microbiota. The output is given as arbitrary bacterial quantities, which for large sample numbers allow for further characterization of the gut microbiota by uni- and multivariate statistical methods....

  11. PCR

    African Journals Online (AJOL)

    Jane

    2011-07-11

    Jul 11, 2011 ... then in age group 31 to 40 years, 2.25% (2/89) CMV DNA were detected by PCR and 0% was recorded in age group of above 40 years. The overall prevalence of human cytomegalovirus (HCMV) infection in 16 ..... genome revisited: Comparison with the chimpanzee cytomegalovirus genome. J. Gen. Virol.

  12. Identification of Multiple Bacteriocins in Enterococcus spp. Using an Enterococcus-Specific Bacteriocin PCR Array

    Directory of Open Access Journals (Sweden)

    Chris Henning

    2015-02-01

    Full Text Available Twenty-two bacteriocin-producing Enterococcus isolates obtained from food and animal sources, and demonstrating activity against Listeria monocytogenes, were screened for bacteriocin-related genes using a bacteriocin PCR array based on known enterococcal bacteriocin gene sequences in the NCBI GenBank database. The 22 bacteriocin-positive (Bac+ enterococci included En. durans (1, En. faecalis (4, En. faecium (12, En. hirae (3, and En. thailandicus (2. Enterocin A (entA, enterocins mr10A and mr10B (mr10AB, and bacteriocin T8 (bacA were the most commonly found structural genes in order of decreasing prevalence. Forty-five bacteriocin genes were identified within the 22 Bac+ isolates, each containing at least one of the screened structural genes. Of the 22 Bac+ isolates, 15 possessed two bacteriocin genes, seven isolates contained three different bacteriocins, and three isolates contained as many as four different bacteriocin genes. These results may explain the high degree of bactericidal activity observed with various Bac+ Enterococcus spp. Antimicrobial activity against wild-type L. monocytogenes and a bacteriocin-resistant variant demonstrated bacteriocins having different modes-of-action. Mixtures of bacteriocins, especially those with different modes-of-action and having activity against foodborne pathogens, such as L. monocytogenes, may play a promising role in the preservation of food.

  13. Cross-platform comparison of SYBR® Green real-time PCR with TaqMan PCR, microarrays and other gene expression measurement technologies evaluated in the MicroArray Quality Control (MAQC study

    Directory of Open Access Journals (Sweden)

    Dial Stacey L

    2008-07-01

    Full Text Available Abstract Background The MicroArray Quality Control (MAQC project evaluated the inter- and intra-platform reproducibility of seven microarray platforms and three quantitative gene expression assays in profiling the expression of two commercially available Reference RNA samples (Nat Biotechnol 24:1115-22, 2006. The tested microarrays were the platforms from Affymetrix, Agilent Technologies, Applied Biosystems, GE Healthcare, Illumina, Eppendorf and the National Cancer Institute, and quantitative gene expression assays included TaqMan® Gene Expression PCR Assay, Standardized (Sta RT-PCR™ and QuantiGene®. The data showed great consistency in gene expression measurements across different microarray platforms, different technologies and test sites. However, SYBR® Green real-time PCR, another common technique utilized by half of all real-time PCR users for gene expression measurement, was not addressed in the MAQC study. In the present study, we compared the performance of SYBR Green PCR with TaqMan PCR, microarrays and other quantitative technologies using the same two Reference RNA samples as the MAQC project. We assessed SYBR Green real-time PCR using commercially available RT2 Profiler™ PCR Arrays from SuperArray, containing primer pairs that have been experimentally validated to ensure gene-specificity and high amplification efficiency. Results The SYBR Green PCR Arrays exhibit good reproducibility among different users, PCR instruments and test sites. In addition, the SYBR Green PCR Arrays have the highest concordance with TaqMan PCR, and a high level of concordance with other quantitative methods and microarrays that were evaluated in this study in terms of fold-change correlation and overlap of lists of differentially expressed genes. Conclusion These data demonstrate that SYBR Green real-time PCR delivers highly comparable results in gene expression measurement with TaqMan PCR and other high-density microarrays.

  14. A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection

    DEFF Research Database (Denmark)

    Hung, Tran Quang; Chin, Wai Hoe; Sun, Yi

    2016-01-01

    technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP......-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP......-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples....

  15. Development of a Multiplex PCR for Discrimination of the TLC:RS1:CTX array of Vibrio cholerae Wave 3 El Tor Strains.

    Science.gov (United States)

    Kim, Eun Jin; Yu, Hyun Jin; Nair, G Balakrish; Kim, Dong Wook

    2016-12-28

    Vibrio cholerae O1 serogroup Wave 3 El Tor strains are presently prevalent worldwide. The Wave 3 El Tor strains contain a TLC:RS1:CTX array on chromosome 1, and no element is integrated on chromosome 2. A multiplex PCR optimized to identify the TLC:RS1:CTX array of Wave 3 strains has been developed in this study. By using eight primers, the multiplex PCR can identify the characteristic CTX and RS1 array of Wave 3 strains from various arrays of strains belonging to other Waves. The four amplified DNA fragments of Wave 3 strains have been cloned in a vector, which could be used as a positive control for the multiplex PCR. This multiplex PCR and the positive control set could be useful tools for rapid recognition of Wave 3 El Tor strains.

  16. A Novel Pressure Indicator for Continuous Flow PCR Chip Using Micro Molded PDMS Pillar Arrays

    National Research Council Canada - National Science Library

    Zhao, Yi; Zhang, Xin

    2005-01-01

    .... Continuous flow PCR chip releases biologists from their laborious exercises. The use of such chip is, however, hindered by costly expense of the syringe pump, which is used to maintain a constant flow rate...

  17. A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection.

    Science.gov (United States)

    Hung, Tran Quang; Chin, Wai Hoe; Sun, Yi; Wolff, Anders; Bang, Dang Duong

    2017-04-15

    Solid-phase PCR (SP-PCR) has become increasingly popular for molecular diagnosis and there have been a few attempts to incorporate SP-PCR into lab-on-a-chip (LOC) devices. However, their applicability for on-line diagnosis is hindered by the lack of sensitive and portable on-chip optical detection technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Digital quantification of gene methylation in stool DNA by emulsion-PCR coupled with hydrogel immobilized bead-array.

    Science.gov (United States)

    Liu, Yunlong; Wu, Haiping; Zhou, Qiang; Song, Qinxin; Rui, Jianzhong; Zou, Bingjie; Zhou, Guohua

    2017-06-15

    Aberrations of gene methylation in stool DNA (sDNA) is an effective biomarker for non-invasive colorectal cancer diagnosis. However, it is challenging to accurately quantitate the gene methylation levels in sDNA due to the low abundance and degradation of sDNA. In this study, a digital quantification strategy was proposed by combining emulsion PCR (emPCR) with hydrogel immobilized bead-array. The assay includes following steps: bisulfite conversion of sDNA, pre-amplification by PCR with specific primers containing 5' universal sequences, emPCR of pre-amplicons with beaded primers to achieve single-molecular amplification and identification of hydrogel embedding beads coated with amplicons. The sensitivity and the specificity of the method are high enough to pick up 0.05% methylated targets from unmethylated DNA background. The successful detection of hypermethylated vimentin gene in clinical stool samples suggests that the proposed method should be a potential tool for non-invasive colorectal cancer screening. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Introducing GUt Low-Density Array (GULDA) - a validated approach for qPCR-based intestinal microbial community analysis

    DEFF Research Database (Denmark)

    Bergström, Anders; Licht, Tine Rask; Wilcks, Andrea

    2012-01-01

    . In the present study, we developed and validated a high-throughput real-time quantitative PCR-based analysis platform, termed 'GUt Low-Density Array' (GULDA). The platform was designed for simultaneous analysis of the change in the abundance of 31 different microbial 16S rRNA gene targets in fecal samples....... To demonstrate the applicability of GULDA, analysis of fecal samples obtained from six healthy infants at both 9 and 18 months of age was performed and showed a significant increase over time of the relative abundance of bacteria belonging to Clostridial cluster IV (Clostridia leptum group) and Bifidobacterium...

  20. High Throughput Sample Preparation and Analysis for DNA Sequencing, PCR and Combinatorial Screening of Catalysis Based on Capillary Array Technique

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yonghua [Iowa State Univ., Ames, IA (United States)

    2000-01-01

    Sample preparation has been one of the major bottlenecks for many high throughput analyses. The purpose of this research was to develop new sample preparation and integration approach for DNA sequencing, PCR based DNA analysis and combinatorial screening of homogeneous catalysis based on multiplexed capillary electrophoresis with laser induced fluorescence or imaging UV absorption detection. The author first introduced a method to integrate the front-end tasks to DNA capillary-array sequencers. protocols for directly sequencing the plasmids from a single bacterial colony in fused-silica capillaries were developed. After the colony was picked, lysis was accomplished in situ in the plastic sample tube using either a thermocycler or heating block. Upon heating, the plasmids were released while chromsomal DNA and membrane proteins were denatured and precipitated to the bottom of the tube. After adding enzyme and Sanger reagents, the resulting solution was aspirated into the reaction capillaries by a syringe pump, and cycle sequencing was initiated. No deleterious effect upon the reaction efficiency, the on-line purification system, or the capillary electrophoresis separation was observed, even though the crude lysate was used as the template. Multiplexed on-line DNA sequencing data from 8 parallel channels allowed base calling up to 620 bp with an accuracy of 98%. The entire system can be automatically regenerated for repeated operation. For PCR based DNA analysis, they demonstrated that capillary electrophoresis with UV detection can be used for DNA analysis starting from clinical sample without purification. After PCR reaction using cheek cell, blood or HIV-1 gag DNA, the reaction mixtures was injected into the capillary either on-line or off-line by base stacking. The protocol was also applied to capillary array electrophoresis. The use of cheaper detection, and the elimination of purification of DNA sample before or after PCR reaction, will make this approach an

  1. A Novel Pressure Indicator for Continuous Flow PCR Chip Using Micro Molded PDMS Pillar Arrays

    National Research Council Canada - National Science Library

    Zhao, Yi; Zhang, Xin

    2005-01-01

    .... A polymeric PCR chip with such pressure indicators is presented. With a much less expense as compared to its conventional peers, this indicator has a wide potential for the use in the laboratories which runs daily activities like sequencing or mutagenesis.

  2. An integrated microfluidic device utilizing dielectrophoresis and multiplex array PCR for point-of-care detection of pathogens.

    Science.gov (United States)

    Cai, Dongyang; Xiao, Meng; Xu, Peng; Xu, Ying-Chun; Du, Wenbin

    2014-10-21

    The early identification of causative pathogens in clinical specimens that require no cultivation is essential for directing evidence-based antimicrobial treatments in resource limited settings. Here, we describe an integrated microfluidic device for the rapid identification of pathogens in complex physiological matrices such as blood. The device was designed and fabricated using SlipChip technologies, which integrated four channels processing independent samples and identifying up to twenty different pathogens. Briefly, diluted whole human blood samples were directly injected into the device for analysis. The pathogens were extracted from the blood by dielectrophoresis, retained in an array of grooves, and identified by multiplex array PCR in nanoliter volumes with end-point fluorescence detection. The universality of the dielectrophoretic separation of pathogens from physiological fluids was evaluated with a panel of clinical isolates covering predominant bacterial and fungal species. Using this system, we simultaneously identified Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli O157:H7 within 3 h. In addition to the prompt diagnosis of bloodstream infections, this method may also be utilized for differentiating microorganisms in contaminated water and environmental samples.

  3. Detection of Her2-overexpressing cancer cells using keyhole shaped chamber array employing a magnetic droplet-handling system.

    Science.gov (United States)

    Okochi, Mina; Koike, Shinji; Tanaka, Masayoshi; Honda, Hiroyuki

    2017-07-15

    An on-chip gene expression analysis compartmentalized in droplets was developed for detection of cancer cells at a single-cell level. The chip consists of a keyhole-shaped reaction chamber with hydrophobic modification employing a magnetic bead-droplet-handling system with a gate for bead separation. Using three kinds of water-based droplets in oil, a droplet with sample cells, a lysis buffer with magnetic beads, and RT-PCR buffer, parallel magnetic manipulation and fusion of droplets were performed using a magnet-handling device containing small external magnet patterns in an array. The actuation with the magnet offers a simple system for droplet manipulation that allows separation and fusion of droplets containing magnetic beads. After reverse transcription and amplification by thermal cycling, fluorescence was obtained for detection of overexpressing genes. For clinical detection of gastric cancer cells in peritoneal washing, the Her2-overexpressing gastric cancer cells spiked within normal cells was detected by gene expression analysis of droplets containing an average of 2.5 cells. Our developed droplet-based cancer detection system manipulated by external magnetic force without pumps or valves offers a simple and flexible set-up for transcriptional detection of cancer cells, and will be greatly advantageous for less-invasive clinical diagnosis and prognostic prediction. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth disease virus and look-alike disease viruses

    Energy Technology Data Exchange (ETDEWEB)

    Hindson, B J; Reid, S M; Baker, B R; Ebert, K; Ferris, N P; Bentley Tammero, L F; Lenhoff, R J; Naraghi-Arani, P; Vitalis, E A; Slezak, T R; Hullinger, P J; King, D P

    2007-07-26

    A high-throughput multiplexed assay was developed for the differential laboratory diagnosis of foot-and-mouth disease virus (FMDV) from viruses which cause clinically similar diseases of livestock. This assay simultaneously screens for five RNA and two DNA viruses using multiplexed reverse transcription PCR (mRT-PCR) amplification coupled with a microsphere hybridization array and flow-cytometric detection. Two of the seventeen primer-probe sets included in this multiplex assay were adopted from previously characterized real-time RT-PCR (rRT-PCR) assays for FMDV. The diagnostic accuracy of the mRT-PCR was evaluated using 287 field samples, including 248 (true positive n= 213, true negative n=34) from suspect cases of foot-and-mouth disease collected from 65 countries between 1965 and 2006 and 39 true negative samples collected from healthy animals. The mRT-PCR assay results were compared with two singleplex rRT-PCR assays, using virus isolation with antigen-ELISA as the reference method. The diagnostic sensitivity of the mRT-PCR assay for FMDV was 93.9% [95% C.I. 89.8-96.4%], compared to 98.1% [95% C.I. 95.3-99.3%] for the two singleplex rRT-PCR assays used in combination. In addition, the assay could reliably differentiate between FMDV and other vesicular viruses such as swine vesicular disease virus and vesicular exanthema of swine virus. Interestingly, the mRT-PCR detected parapoxvirus (n=2) and bovine viral diarrhea virus (n=2) in clinical samples, demonstrating the screening potential of this mRT-PCR assay to identify viruses in FMDV-negative material not previously recognized using focused single-target rRT-PCR assays.

  5. Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth and look-alike disease viruses

    Energy Technology Data Exchange (ETDEWEB)

    Hindson, B J; Baker, B R; Bentley Tammero, L F; Lenhoff, R J; Naraghi-Arani, P; Vitalis, E A; Slezak, T R; Hullinger, P J; Reid, S M; Ebert, K; Ferris, N P; King, D P

    2007-09-18

    A high-throughput multiplexed assay (Multiplex Version 1.0) was developed for the differential laboratory diagnosis of foot-and-mouth disease virus (FMDV) from viruses which cause clinically similar diseases of livestock. This assay simultaneously screens for five RNA and two DNA viruses using multiplexed reverse transcription PCR (mRT-PCR) amplification coupled with a microsphere hybridization array and flow-cytometric detection. Two of the seventeen primer-probe sets included in this multiplex assay were adopted from previously characterized real-time RT-PCR (rRT-PCR) assays for FMDV. The diagnostic accuracy of the mRT-PCR was evaluated using 287 field samples, including 248 (true positive n= 213, true negative n=34) from suspect cases of foot-and-mouth disease collected from 65 countries between 1965 and 2006 and 39 true negative samples collected from healthy animals. The mRT-PCR assay results were compared with two singleplex rRT-PCR assays, using virus isolation with antigen-ELISA as the reference method. The diagnostic sensitivity of the mRT-PCR assay for FMDV was 93.9% [95% C.I. 89.8-96.4%], compared to 98.1% [95% C.I. 95.3-99.3%] for the two singleplex rRTPCR assays used in combination. In addition, the assay could reliably differentiate between FMDV and other vesicular viruses such as swine vesicular disease virus and vesicular exanthema of swine virus. Interestingly, the mRT-PCR detected parapoxvirus (n=2) and bovine viral diarrhea virus (n=2) in clinical samples, demonstrating the screening potential of this mRT-PCR assay to identify viruses in FMDV-negative material not previously recognized using focused single-target rRT-PCR assays.

  6. Comparison of LCD array and IS6110-PCR with conventional techniques for detection of Mycobacterium bovis isolated from Egyptian cattle and Buffaloes.

    Science.gov (United States)

    Zahran, Rasha Nabil; El Behiry, Ayman; Marzouk, Eman; Askar, Tamer

    2014-09-01

    Bovine tuberculosis is a chronic bacterial and major infectious disease of cattle and buffaloes caused by Mycobacterium bovis. Rapid diagnosis of bovine tuberculosis is considered one of the cornerstones for worldwide control as it permits early epidemiological and therapeutic interventions. Therefore, this study was designed to evaluate conventional techniques (tuberculin test, Ziehl Neelsen staining and culturing) in comparison with proven molecular laboratory techniques (LCD array and IS6110 PCR) for identification of Bovine tuberculosis. A total of 902 Egyptian animals (480 buffaloes and 422 cattle) were examined by tuberculin test, and the positive reactors were slaughtered. Tissue samples were collected for staining as well as culturing. Moreover, LCD array and PCR using IS6110 on DNA extracted from tissue and culture samples were carried out for molecular identification of M. bovis. According to the results, the tuberculin positive cases for cattle and buffaloes were 2.14% (9 cases) and 5.62% (27 cases), respectively. After post-mortem examination, the prevalence of tuberculin positive cases with visible lesions was 88.9% for cattle and 14.8% for buffaloes. Alternatively, these percentages were 11.1% and 85.2% for cattle and buffalo carcasses with non-visible lesions. The percentage of cattle and buffaloes showing positive culture was 88.9% and 62.9%, respectively. This percentage was 69.5% after staining with Ziehl Neelsen. In contrast, LCD array and IS6110 were 100%, confirming the isolation results. In conclusion, LCD array depending on 16S RNA and DNA hybridization with specific probes for detection of M. bovis are rapid, sensitive and labor-saving when combined with IS6110-PCR. Copyright © 2014 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  7. Validation of GUt Low Density Array (GULDA), a novel qPCR approach to the study of the intestinal microbial ecosystem

    DEFF Research Database (Denmark)

    Bergström, Anders; Licht, Tine Rask; Bahl, Martin Iain

    Causal relationships between the vast numbers of bacterial species present in the human intestines contain a lot of potential information on the regulation of the gut in the healthy as well as in diseased states. Based on the hypothesis that the human gut microbiota constitutes a dynamic ecosyste...... one standard qPCR program is used for ~40 primer sets, validation is important. We present here strategies involved in verification of GULDA as a valid tool for analysis of the human gut microbiota......., interesting correlations between the presences of the given species should exist at any time. In order to analyze this, we have developed GULDA, a cheap, flexible, reliable and high throughput qPCR-based gut low-density array (GULDA), which simultaneously gives the quantities of approximately 40 different...

  8. Characterization of the infant gut microbiota in a cohort of 330 Danish children at 9, 18 and 36 months by quantitative PCR array (GULDA) analysis

    DEFF Research Database (Denmark)

    Bergström, Anders; Skov, Thomas; Bahl, Martin Iain

    (the so called SKOT-cohort), sampled at 9, 18 and 36 months after birth. The resulting data together with previously determined nutritional and anthropometrical parameters were used as input for multivariate statistics. We found significant changes in the composition of the gut microbiota between 9...... for microbial community characterization, which here provides new insights into the interactions between the gut microbiota, diet and physiology in infants.......We have developed a qPCR-based array (GUt Low Density Array, GULDA), which simultaneously determine the relative abundance of >30 different bacterial 16S rRNA gene targets in a given DNA-sample covering selected phylogenetic levels. GULDA was applied to fecal DNA from 330 healthy Danish infants...

  9. Eight-plex PCR and liquid-array detection of bacterial and viral pathogens in cerebrospinal fluid from patients with suspected meningitis

    DEFF Research Database (Denmark)

    Bøving, Mette Kusk; Pedersen, Lisbeth Nørum; Møller, Jens Kjølseth

    2009-01-01

    We here report on the development of a novel multiplex PCR with product detection in a Luminex 100 suspension array system. The assay covers the nine most important bacterial and viral pathogens found in Danish meningitis patients. The microorganisms include Neisseria meningitidis, Streptococcus...... and culture of cerebrospinal fluid (CSF) samples in a routine diagnostic setting, and results can be available within 1 workday. The method is suitable for use for the initial screening and identification of nine important microorganisms in CSF samples from patients with suspected meningitis. Compared...

  10. A hybrid qPCR/SNP array approach allows cost efficient assessment of KIR gene copy numbers in large samples.

    Science.gov (United States)

    Pontikos, Nikolas; Smyth, Deborah J; Schuilenburg, Helen; Howson, Joanna M M; Walker, Neil M; Burren, Oliver S; Guo, Hui; Onengut-Gumuscu, Suna; Chen, Wei-Min; Concannon, Patrick; Rich, Stephen S; Jayaraman, Jyothi; Jiang, Wei; Traherne, James A; Trowsdale, John; Todd, John A; Wallace, Chris

    2014-04-11

    Killer Immunoglobulin-like Receptors (KIRs) are surface receptors of natural killer cells that bind to their corresponding Human Leukocyte Antigen (HLA) class I ligands, making them interesting candidate genes for HLA-associated autoimmune diseases, including type 1 diabetes (T1D). However, allelic and copy number variation in the KIR region effectively mask it from standard genome-wide association studies: single nucleotide polymorphism (SNP) probes targeting the region are often discarded by standard genotype callers since they exhibit variable cluster numbers. Quantitative Polymerase Chain Reaction (qPCR) assays address this issue. However, their cost is prohibitive at the sample sizes required for detecting effects typically observed in complex genetic diseases. We propose a more powerful and cost-effective alternative, which combines signals from SNPs with more than three clusters found in existing datasets, with qPCR on a subset of samples. First, we showed that noise and batch effects in multiplexed qPCR assays are addressed through normalisation and simultaneous copy number calling of multiple genes. Then, we used supervised classification to impute copy numbers of specific KIR genes from SNP signals. We applied this method to assess copy number variation in two KIR genes, KIR3DL1 and KIR3DS1, which are suitable candidates for T1D susceptibility since they encode the only KIR molecules known to bind with HLA-Bw4 epitopes. We find no association between KIR3DL1/3DS1 copy number and T1D in 6744 cases and 5362 controls; a sample size twenty-fold larger than in any previous KIR association study. Due to our sample size, we can exclude odds ratios larger than 1.1 for the common KIR3DL1/3DS1 copy number groups at the 5% significance level. We found no evidence of association of KIR3DL1/3DS1 copy number with T1D, either overall or dependent on HLA-Bw4 epitope. Five other KIR genes, KIR2DS4, KIR2DL3, KIR2DL5, KIR2DS5 and KIR2DS1, in high linkage disequilibrium with

  11. Altered Gene Expression by Low-Dose Arsenic Exposure in Humans and Cultured Cardiomyocytes: Assessment by Real-Time PCR Arrays

    Directory of Open Access Journals (Sweden)

    Judy Mumford

    2011-06-01

    Full Text Available Chronic arsenic exposure results in higher risk of skin, lung, and bladder cancer, as well as cardiovascular disease and diabetes. The purpose of this study was to investigate the effects on expression of selected genes in the blood lymphocytes from 159 people exposed chronically to arsenic in their drinking water using a novel RT-PCR TaqMan low-density array (TLDA. We found that expression of tumor necrosis factor-α (TNF-α, which activates both inflammation and NF-κB-dependent survival pathways, was strongly associated with water and urinary arsenic levels. Expression of KCNA5, which encodes a potassium ion channel protein, was positively associated with water and toe nail arsenic levels. Expression of 2 and 11 genes were positively associated with nail and urinary arsenic, respectively. Because arsenic exposure has been reported to be associated with long QT intervals and vascular disease in humans, we also used this TLDA for analysis of gene expression in human cardiomyocytes exposed to arsenic in vitro. Expression of the ion-channel genes CACNA1, KCNH2, KCNQ1 and KCNE1 were down-regulated by 1-mM arsenic. Alteration of some common pathways, including those involved in oxidative stress, inflammatory signaling, and ion-channel function, may underlay the seemingly disparate array of arsenic-associated diseases, such as cancer, cardiovascular disease, and diabetes.

  12. A high-throughput open-array qPCR gene panel to identify housekeeping genes suitable for myometrium and leiomyoma expression analysis.

    Science.gov (United States)

    Almeida, Teresa A; Quispe-Ricalde, Antonieta; Montes de Oca, Francisco; Foronda, Pilar; Hernández, Mariano M

    2014-07-01

    To evaluate 51 different housekeeping genes for their use as internal standards in myometrial and matched leiomyoma samples in proliferative and secretory phases. RNA from 6 myometrium and matched leiomyoma samples was obtained from pre-menopausal women who underwent hysterectomy. Reverse-transcription and real-time quantitative PCR were achieved using TaqMan high-density open-array human endogenous control panel. Expression stability of 51 candidate genes was determined by GeNorm and NormFinder softwares. We identified 10 housekeeping genes, ARF1, MRPL19, FBXW2, PUM1, UBE2D2, EIF2B1, HPRT1, GUSB, ALAS1, and TRIM27, as the best set of normalization genes for comparing relative expression between leiomyoma and myometrium samples in proliferative and secretory phases. Adequate reference genes for accurate normalization are essential to compare gene expression between leiomyoma and myometrial samples. Ideal housekeeping genes must have stable expression patterns regardless of the sample type and menstrual cycle phase. In this study, we propose a set of 10 candidate genes with greater expression stability than those housekeeping genes commonly used in leiomyoma and myometrium tissues. Their use will improve the sensitivity and specificity of the gene expression analysis in these tissues. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. Detection of siRNA Mediated Target mRNA Cleavage Activities in Human Cells by a Novel Stem-Loop Array RT-PCR Analysis

    Science.gov (United States)

    2016-09-07

    of input synthetic RNU44 ducts were analyzed by agarose gel electrophoresis (B). (C) Dynamic range of the SL- by using SLA-RT-PCR assay and SL-RT-PCR...by agarose gel electrophoresis (upper panel) and relative band intensity was shown in lower panel. J. Lin et al. / Biochemistry and Biophysics...by polyacrylamide gel electrophoresis (PAGE). The purified RUN44 RNA was used as a RNA template to evaluate SLA-RT-PCR for detection and verification

  14. Accurate quantification of microorganisms in PCR-inhibiting environmental DNA extracts by a novel Internal Amplification Control approach using Biotrove OpenArrays

    NARCIS (Netherlands)

    Van Doorn, R.; Klerks, M.; van Gent-Pelzer, M.; Speksnijder, A.G.C.L.; Kowalchuk, G.A.; Schoen, C.D.

    2009-01-01

    PCR-based detection assays are prone to inhibition by substances present in environmental samples, thereby potentially leading to inaccurate target quantification or false-negative results. Internal amplification controls (IACs) have been developed to help alleviate this problem but are generally

  15. Differential expression of cellular microRNAs in HPV-11 transfected cells. An analysis by three different array platforms and qRT-PCR

    DEFF Research Database (Denmark)

    Dreher, Anita; Rossing, Maria; Kaczkowski, Bogumil

    2010-01-01

    . The changes of cellular microRNAs by HPV-11 gene expression were investigated in a cell culture model of HaCaT cells transfected with HPV-11, with the goal of understanding which cellular processes were affected by the virus. Human microRNA profiling was conducted on three different array platform systems...

  16. Comparison of In-House Multiplex Real Time PCR, Diagcor GenoFlow HPV Array Test and INNO-LiPA HPV Genotyping Extra Assays with LCD- Array Kit for Human Papillomavirus Genotyping in Cervical Liquid Based Cytology Specimens and Genital Lesions in Tehran, Iran.

    Science.gov (United States)

    Sohrabi, Amir; Rahnamaye-Farzami, Marjan; Mirab-Samiee, Siamak; Mahdavi, Saeed; Babaei, Monireh

    2016-01-01

    Human papillomavirus is a major etiologic agent for some human common cancers. Cervical precancer and cancer is the most prevalent dysplasia by HPV genotypes. Various rapid and sensitive methods have been developed into readily HPV genotyping. In the present study, we compared the performance of Real Time PCR, GenoFlow HPV Array, and INNO-LiPA HPV Genotyping Extra Assays with LCD- Array. From 108 cervical samples, HPV was detected in 33 women (30.55%). Among detected HPV genotypes, HPV 6 and 11 were dominant genotypes. Comparing these methods revealed that for Real Time PCR, Genoflow, and INNO-LiPA in comparison with LCD Array, sensitivity and specificity were 94.2%, 93%; 76.7%, 93%; 64%, 96.5%, respectively. Overall, accuracy and precision of these methods were more than 80% and 90%, respectively. It seems that these methods are reliable and suitable for detection and genotyping of HPVs in cervical disorders and other dysplasia associated with human papillomaviruses.

  17. Performance of the digene LQ, RH and PS HPVs genotyping systems on clinical samples and comparison with HC2 and PCR-based Linear Array

    Directory of Open Access Journals (Sweden)

    Godínez Jose M

    2011-11-01

    Full Text Available Abstract Background Certain Human Papillomaviruses (HPVs are the infectious agents involved in cervical cancer development. Detection of HPVs DNA is part of the cervical cancer screening protocols and HPVs genotyping has been proposed for its inclusion in these preventive programs. The aim of this study was to evaluate three novel genotyping tests, namely Qiagen LQ, RH and PS, in clinical samples with and without abnormalities. For this, 305 cervical samples were processed and the results of the evaluated techniques were compared with those obtained in the HPVs diagnostic process in our lab, by using HC2 and Linear Array (LA technologies. Results The concordances and kappa statistics (k for each technique compared with HC2 were 98.69% (k = 0.94 for LQ, 98.03% (k = 0.91 for RH and 91.80% (k = 0.82 for PS. There was a very good agreement in HPVs type-specific concordance for the most prevalent types HPV16 (kappa range = 0.83-0.90, HPV18 (k.r.= 0.74-0.80 and HPV45 (k.r.= 0.82-0.90. Conclusions The three tests showed an overall good concordance for HPVs detection when compared with HR-HC2 system. LQ and RH rendered lower detection rate for multiple infections than LA genotyping. However, our understanding of the clinical significance of multiple HPVs infections is still incomplete and therefore the relevance of the lower ability to detect multiple infections needs to be evaluated.

  18. Replica amplification of nucleic acid arrays

    Science.gov (United States)

    Church, George M.; Mitra, Robi D.

    2010-08-31

    Disclosed are improved methods of making and using immobilized arrays of nucleic acids, particularly methods for producing replicas of such arrays. Included are methods for producing high density arrays of nucleic acids and replicas of such arrays, as well as methods for preserving the resolution of arrays through rounds of replication. Also included are methods which take advantage of the availability of replicas of arrays for increased sensitivity in detection of sequences on arrays. Improved methods of sequencing nucleic acids immobilized on arrays utilizing single copies of arrays and methods taking further advantage of the availability of replicas of arrays are disclosed. The improvements lead to higher fidelity and longer read lengths of sequences immobilized on arrays. Methods are also disclosed which improve the efficiency of multiplex PCR using arrays of immobilized nucleic acids.

  19. Magnetic arrays

    Science.gov (United States)

    Trumper, D.L.; Kim, W.; Williams, M.E.

    1997-05-20

    Electromagnet arrays are disclosed which can provide selected field patterns in either two or three dimensions, and in particular, which can provide single-sided field patterns in two or three dimensions. These features are achieved by providing arrays which have current densities that vary in the windings both parallel to the array and in the direction of array thickness. 12 figs.

  20. Magnetic arrays

    Energy Technology Data Exchange (ETDEWEB)

    Trumper, David L. (Plaistow, NH); Kim, Won-jong (Cambridge, MA); Williams, Mark E. (Pelham, NH)

    1997-05-20

    Electromagnet arrays which can provide selected field patterns in either two or three dimensions, and in particular, which can provide single-sided field patterns in two or three dimensions. These features are achieved by providing arrays which have current densities that vary in the windings both parallel to the array and in the direction of array thickness.

  1. Assessment of the real-time PCR and different digital PCR platforms for DNA quantification.

    Science.gov (United States)

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2016-01-01

    Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been done, and the 37 K array on the Biomark system has also not been evaluated in terms of linearity, analytical sensitivity and limit of quantification. Here, a first assessment of qPCR, the QX100 system and both arrays of the Biomark system was performed with plasmid and genomic DNA from human cytomegalovirus. With use of PCR components that alter the efficiency of qPCR, each dPCR platform demonstrated consistent copy-number estimations, which indicates the high resilience of dPCR. Two approaches, one considering the total reaction volume and the other considering the effective reaction size, were used to assess linearity, analytical sensitivity and variability. When the total reaction volume was considered, the best performance was observed with qPCR, followed by the QX100 system and the Biomark system. In contrast, when the effective reaction size was considered, all three platforms showed almost equal limits of detection and variability. Although dPCR might not always be more appropriate than qPCR for quantification of low copy numbers, dPCR is a suitable method for robust and reproducible quantification of viral DNA, and a promising technology for the higher-order reference measurement method.

  2. Design Considerations for Array CGH to OligonucleotideArrays

    Energy Technology Data Exchange (ETDEWEB)

    Baldocchi, R.A.; Glynne, R.J.; Chin, K.; Kowbel, D.; Collins, C.; Mack, D.H.; Gray, J.W.

    2005-03-04

    Background: Representational oligonucleotide microarray analysis has been developed for detection of single nucleotide polymorphisms and/or for genome copy number changes. In this process, the intensity of hybridization to oligonucleotides arrays is increased by hybridizing a polymerase chain reaction (PCR)-amplified representation of reduced genomic complexity. However, hybridization to some oligonucleotides is not sufficiently high to allow precise analysis of that portion of the genome. Methods: In an effort to identify aspects of oligonucleotide hybridization affecting signal intensity, we explored the importance of the PCR product strand to which each oligonucleotide is homologous and the sequence of the array oligonucleotides. We accomplished this by hybridizing multiple PCR-amplified products to oligonucleotide arrays carrying two sense and two antisense 50-mer oligonucleotides for each PCR amplicon. Results: In some cases, hybridization intensity depended more strongly on the PCR amplicon strand (i.e., sense vs. antisense) than on the detection oligonucleotide sequence. In other cases, the oligonucleotide sequence seemed to dominate. Conclusion: Oligonucleotide arrays for analysis of DNA copy number or for single nucleotide polymorphism content should be designed to carry probes to sense and antisense strands of each PCR amplicon to ensure sufficient hybridization and signal intensity.

  3. Array capabilities and future arrays

    International Nuclear Information System (INIS)

    Radford, D.

    1993-01-01

    Early results from the new third-generation instruments GAMMASPHERE and EUROGAM are confirming the expectation that such arrays will have a revolutionary effect on the field of high-spin nuclear structure. When completed, GAMMASHPERE will have a resolving power am order of magnitude greater that of the best second-generation arrays. When combined with other instruments such as particle-detector arrays and fragment mass analysers, the capabilites of the arrays for the study of more exotic nuclei will be further enhanced. In order to better understand the limitations of these instruments, and to design improved future detector systems, it is important to have some intelligible and reliable calculation for the relative resolving power of different instrument designs. The derivation of such a figure of merit will be briefly presented, and the relative sensitivities of arrays currently proposed or under construction presented. The design of TRIGAM, a new third-generation array proposed for Chalk River, will also be discussed. It is instructive to consider how far arrays of Compton-suppressed Ge detectors could be taken. For example, it will be shown that an idealised open-quote perfectclose quotes third-generation array of 1000 detectors has a sensitivity an order of magnitude higher again than that of GAMMASPHERE. Less conventional options for new arrays will also be explored

  4. SNP Arrays

    Directory of Open Access Journals (Sweden)

    Jari Louhelainen

    2016-10-01

    Full Text Available The papers published in this Special Issue “SNP arrays” (Single Nucleotide Polymorphism Arrays focus on several perspectives associated with arrays of this type. The range of papers vary from a case report to reviews, thereby targeting wider audiences working in this field. The research focus of SNP arrays is often human cancers but this Issue expands that focus to include areas such as rare conditions, animal breeding and bioinformatics tools. Given the limited scope, the spectrum of papers is nothing short of remarkable and even from a technical point of view these papers will contribute to the field at a general level. Three of the papers published in this Special Issue focus on the use of various SNP array approaches in the analysis of three different cancer types. Two of the papers concentrate on two very different rare conditions, applying the SNP arrays slightly differently. Finally, two other papers evaluate the use of the SNP arrays in the context of genetic analysis of livestock. The findings reported in these papers help to close gaps in the current literature and also to give guidelines for future applications of SNP arrays.

  5. PCR-RFLP

    African Journals Online (AJOL)

    2012-03-30

    Mar 30, 2012 ... The. cpDNA PCR-RFLP based genetic distance (GD) among 30 tea accessions ranged from 0 to 0.071, with the mean of 0.049. This study suggests that the optimization system was suitable for PCR-RFLP analysis of. cpDNA in tea. Key words: Camellia sinensis, PCR-RFLP, chloroplast DNA, establishment.

  6. Filter arrays

    Energy Technology Data Exchange (ETDEWEB)

    Page, Ralph H.; Doty, Patrick F.

    2017-08-01

    The various technologies presented herein relate to a tiled filter array that can be used in connection with performance of spatial sampling of optical signals. The filter array comprises filter tiles, wherein a first plurality of filter tiles are formed from a first material, the first material being configured such that only photons having wavelengths in a first wavelength band pass therethrough. A second plurality of filter tiles is formed from a second material, the second material being configured such that only photons having wavelengths in a second wavelength band pass therethrough. The first plurality of filter tiles and the second plurality of filter tiles can be interspersed to form the filter array comprising an alternating arrangement of first filter tiles and second filter tiles.

  7. CODEHOP PCR and CODEHOP PCR primer design.

    Science.gov (United States)

    Staheli, Jeannette P; Boyce, Richard; Kovarik, Dina; Rose, Timothy M

    2011-01-01

    While PCR primer design for the amplification of known sequences is usually quite straightforward, the design, and successful application of primers aimed at the detection of as yet unknown genes is often not. The search for genes that are presumed to be distantly related to a known gene sequence, such as homologous genes in different species, paralogs in the same genome, or novel pathogens in diverse hosts, often turns into the proverbial search for the needle in the haystack. PCR-based methods commonly used to address this issue involve the use of either consensus primers or degenerate primers, both of which have significant shortcomings regarding sensitivity and specificity. We have developed a novel primer design approach that diminishes these shortcomings and instead takes advantage of the strengths of both consensus and degenerate primer designs, by combining the two concepts into a Consensus-Degenerate Hybrid Oligonucleotide Primer (CODEHOP) approach. CODEHOP PCR primers contain a relatively short degenerate 3' core and a 5' nondegenerate clamp. The 3' degenerate core consists of a pool of primers containing all possible codons for a 3-4 aminoacid motif that is highly conserved in multiply aligned sequences from known members of a protein family. Each primer in the pool also contains a single 5' nondegenerate nucleotide sequence derived from a codon consensus across the aligned aminoacid sequences flanking the conserved motif. During the initial PCR amplification cycles, the degenerate core is responsible for specific binding to sequences encoding the conserved aminoacid motif. The longer consensus clamp region serves to stabilize the primer and allows the participation of all primers in the pool in the efficient amplification of products during later PCR cycles. We have developed an interactive web site and algorithm (iCODEHOP) for designing CODEHOP PCR primers from multiply aligned protein sequences, which is freely available online. Here, we describe the

  8. Soft colloidal probes for AFM force measurements between water droplets in oil

    KAUST Repository

    Vakarelski, Ivan Uriev

    2014-11-01

    Here we introduce an extension of the atomic force microscopy (AFM) colloidal probe technique, as a simple and reliable experimental approach to measure the interaction forces between small water droplets (~80-160. μm) dispersed in oil. Small water droplets are formed by capillary breakup of a microscale water jet in air, which is forced out of a fine capillary nozzle, and deposited on a superhydrophobic substrate immersed in tetradecane oil medium. In these conditions the water droplets are very loosely attached to the superhydrophobic substrate and are easily picked up with a hydrophobic AFM cantilever to form a soft colloidal probe. Sample force measurements are conducted to demonstrate the capability of the technique.

  9. External PCR, ASN's decision

    International Nuclear Information System (INIS)

    Anon.

    2012-01-01

    The French law imposes in some situations the presence of a person skilled in radiation protection (PCR). This article describes the cases when this person must belong to the staff of the enterprise or when this person may be sub-contracted. For instance in most nuclear facilities the PCR must be on the payroll, for enterprises dedicated to nuclear transport the PCR's job can be sub-contracted. A decision given by the ASN (French Nuclear Safety Authority) sets the minimal requests (in terms of training, job contract, activities) of the sub-contracted PCR. (A.C.)

  10. Molecular Subtyping of Salmonella Typhimurium with Multiplex Oligonucleotide Ligation-PCR (MOL-PCR).

    Science.gov (United States)

    Wuyts, Véronique; Mattheus, Wesley; Roosens, Nancy H C; Marchal, Kathleen; Bertrand, Sophie; De Keersmaecker, Sigrid C J

    2017-01-01

    A multiplex oligonucleotide ligation-PCR (MOL-PCR) assay is a valuable high-throughput technique for the detection of bacteria and viruses, for characterization of pathogens and for diagnosis of genetic diseases, as it allows one to combine different types of molecular markers in a high-throughput multiplex assay. A MOL-PCR assay starts with a multiplex oligonucleotide ligation reaction for detection of the molecular marker, followed by a singleplex PCR for signal amplification and analysis of the MOL-PCR products on a Luminex platform. This last step occurs through a liquid bead suspension array in which the MOL-PCR products are hybridized to MagPlex-TAG beads.In this chapter, we describe the complete procedure for a MOL-PCR assay for subtyping of Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) and its monophasic variant S. 1,4[5],12:i:- from DNA isolation through heat lysis up to data interpretation through a Gödel Prime Product. The subtyping assay consists of 50 discriminative molecular markers and two internal positive control markers divided over three MOL-PCR assays.

  11. Tomographic array

    International Nuclear Information System (INIS)

    1976-01-01

    The configuration of a tomographic array in which the object can rotate about its axis is described. The X-ray detector is a cylindrical screen perpendicular to the axis of rotation. The X-ray source has a line-shaped focus coinciding with the axis of rotation. The beam is fan-shaped with one side of this fan lying along the axis of rotation. The detector screen is placed inside an X-ray image multiplier tube

  12. Tomographic array

    International Nuclear Information System (INIS)

    1976-01-01

    A tomographic array with the following characteristics is described. An X-ray screen serving as detector is placed before a photomultiplier tube which itself is placed in front of a television camera connected to a set of image processors. The detector is concave towards the source and is replacable. Different images of the object are obtained simultaneously. Optical fibers and lenses are used for transmission within the system

  13. Molecular diagnostic PCR handbook

    International Nuclear Information System (INIS)

    Viljoen, G.J.; Crowther, J.R.; Nel, L.H.

    2005-01-01

    The uses of nucleic acid-directed methods have increased significantly in the past five years and have made important contributions to disease control country programmes for improving national and international trade. These developments include the more routine use of PCR as a diagnostic tool in veterinary diagnostic laboratories. However, there are many problems associated with the transfer and particularly, the application of this technology. These include lack of consideration of: the establishment of quality-assured procedures, the required set-up of the laboratory and the proper training of staff. This can lead to a situation where results are not assured. This book gives a comprehensive account of the practical aspects of PCR and strong consideration is given to ensure its optimal use in a laboratory environment. This includes the setting-up of a PCR laboratory; Good Laboratory Practice and standardised PCR protocols to detect animal disease pathogens. Examples of Standard Operating Procedures as used in individual specialist laboratories and an outline of training materials necessary for PCR technology transfer are presented. The difficulties, advantages and disadvantages in PCR applications are explained and placed in context with other test systems. Emphasis is placed on the use of PCR for detection of pathogens, with a particular focus on diagnosticians and scientists from the developing world. It is hoped that this book will enable readers from various disciplines and levels of expertise to better judge the merits of PCR and to increase their skills and knowledge in order to assist in a more logical, efficient and assured use of this technology

  14. Poisson Plus Quantification for Digital PCR Systems.

    Science.gov (United States)

    Majumdar, Nivedita; Banerjee, Swapnonil; Pallas, Michael; Wessel, Thomas; Hegerich, Patricia

    2017-08-29

    Digital PCR, a state-of-the-art nucleic acid quantification technique, works by spreading the target material across a large number of partitions. The average number of molecules per partition is estimated using Poisson statistics, and then converted into concentration by dividing by partition volume. In this standard approach, identical partition sizing is assumed. Violations of this assumption result in underestimation of target quantity, when using Poisson modeling, especially at higher concentrations. The Poisson-Plus Model accommodates for this underestimation, if statistics of the volume variation are well characterized. The volume variation was measured on the chip array based QuantStudio 3D Digital PCR System using the ROX fluorescence level as a proxy for effective load volume per through-hole. Monte Carlo simulations demonstrate the efficacy of the proposed correction. Empirical measurement of model parameters characterizing the effective load volume on QuantStudio 3D Digital PCR chips is presented. The model was used to analyze digital PCR experiments and showed improved accuracy in quantification. At the higher concentrations, the modeling must take effective fill volume variation into account to produce accurate estimates. The extent of the difference from the standard to the new modeling is positively correlated to the extent of fill volume variation in the effective load of your reactions.

  15. Coupling in reflector arrays

    DEFF Research Database (Denmark)

    Appel-Hansen, Jørgen

    1968-01-01

    In order to reduce the space occupied by a reflector array, it is desirable to arrange the array antennas as close to each other as possible; however, in this case coupling between the array antennas will reduce the reflecting properties of the reflector array. The purpose of the present communic......In order to reduce the space occupied by a reflector array, it is desirable to arrange the array antennas as close to each other as possible; however, in this case coupling between the array antennas will reduce the reflecting properties of the reflector array. The purpose of the present...

  16. PCR in forensic genetics

    DEFF Research Database (Denmark)

    Morling, Niels

    2009-01-01

    Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more...... and more advanced, special investigations in cases concerning crime, paternity, relationship, disaster victim identification etc. The present review gives an update on the use of DNA investigations in forensic genetics....

  17. One-stop polymerase chain reaction (PCR): An improved PCR ...

    African Journals Online (AJOL)

    -cycling steps to visualize amplicons, decelerating PCR sample processing and result calling. “One-stop PCR” was developed by including both the loading buffer and nontoxic staining dye within a single PCR tube, allowing direct loading and ...

  18. One-stop polymerase chain reaction (PCR): An improved PCR ...

    African Journals Online (AJOL)

    Yomi

    2011-12-21

    Dec 21, 2011 ... improved PCR method with speedy operation and ... novel PCR method is desired to compatibilize Taq DNA .... as template. A 20 ul traditional PCR mixture included 10×PCR reaction buffer, 2 µl, 40 mM dNTP (10mM each), 0.5 µl; Taq DNA polymerase, 0.2 µl (1 unit), 0.5 µl forward and reverse primer mix ...

  19. Enhancement of PCR Detection Limit by Single-Tube Restriction Endonuclease-PCR (RE-PCR).

    Science.gov (United States)

    Datta, Sibnarayan; Budhauliya, Raghvendra; Chatterjee, Soumya; Vanlalhmuaka; Veer, Vijay; Chakravarty, Runu

    2016-06-01

    Polymerase chain reaction (PCR) is widely used in biological research and diagnostics because of its high sensitivity and specificity. However, the sensitivity of PCR is strongly influenced by topological characteristics of the template. Supercoiled templates are known to inhibit PCR, whereas linearized forms of the same supercoiled templates facilitate PCR. This study was conducted to compare the PCR efficiency of circular supercoiled DNA templates to their restriction endonuclease (RE)-mediated linearized forms. Additionally, we also evaluated the possibility of RE digestion of the circular supercoiled templates within the complete PCR buffer. Following a systematic approach, we demonstrated that circular supercoiled templates could be efficiently linearized by RE in the complete PCR buffer itself. This allowed linearization of circular supercoiled templates and their subsequent amplification in the PCR buffer in a single-tube format. Using this extremely simple RE-PCR approach, we documented up to tenfold increases in detection efficiency of PCR with two different circular supercoiled templates of clinical origin, including an international calibration standard. This inexpensive and easy approach to increasing PCR sensitivity can be easily adapted to any standard PCR protocol aimed at amplifying circular supercoiled genomes. Apart from its application in the development of sensitive clinical diagnostic PCR assays for a large number of organisms, this method could also prove to be very useful in simplifying the existing protocols for other applications where pre-PCR restriction digestion is required, such as mutation detection, genotyping, and selective template amplification.

  20. PCR, exit stage left ...

    CERN Multimedia

    2004-01-01

    The Prevessin Control Room during LEP's start up in 1989. The Prévessin Control Room (PCR) was recently engulfed in a wave of nostalgia. The PCR, scene of some of the greatest moments in CERN's history, is being dismantled to prepare for a complete overhaul. In February 2006, a new combined control centre for all the accelerators will open its doors on the same site, together with a new building currently under construction (see Bulletin issue 27/2004 of 28 June 2004). This marks the end of an important chapter in CERN's history. The Prévessin Control Room saw its first momentous event 28 years ago when the 400 GeV beam for the SPS was commissioned in the presence of Project Leader John Adams. It was also here that the first proton-antiproton collisions were observed, in 1981. Eight years later, in 1989, operators and directors alike jumped for joy at the announcement of the first electron-positron collisions at the start up of LEP, the biggest accelerator in the world. Today the 80 terminals and PCs have b...

  1. Fiber Laser Array

    National Research Council Canada - National Science Library

    Simpson, Thomas

    2002-01-01

    ...., field-dependent, loss within the coupled laser array. During this program, Jaycor focused on the construction and use of an experimental apparatus that can be used to investigate the coherent combination of an array of fiber lasers...

  2. A Novel High-Throughput Method for Molecular Detection of Human Pathogenic Viruses Using a Nanofluidic Real-Time PCR System.

    Science.gov (United States)

    Coudray-Meunier, Coralie; Fraisse, Audrey; Martin-Latil, Sandra; Delannoy, Sabine; Fach, Patrick; Perelle, Sylvie

    2016-01-01

    Human enteric viruses are recognized as the main causes of food- and waterborne diseases worldwide. Sensitive and quantitative detection of human enteric viruses is typically achieved through quantitative RT-PCR (RT-qPCR). A nanofluidic real-time PCR system was used to develop novel high-throughput methods for qualitative molecular detection (RT-qPCR array) and quantification of human pathogenic viruses by digital RT-PCR (RT-dPCR). The performance of high-throughput PCR methods was investigated for detecting 19 human pathogenic viruses and two main process controls used in food virology. The conventional real-time PCR system was compared to the RT-dPCR and RT-qPCR array. Based on the number of genome copies calculated by spectrophotometry, sensitivity was found to be slightly better with RT-qPCR than with RT-dPCR for 14 viruses by a factor range of from 0.3 to 1.6 log10. Conversely, sensitivity was better with RT-dPCR than with RT-qPCR for seven viruses by a factor range of from 0.10 to 1.40 log10. Interestingly, the number of genome copies determined by RT-dPCR was always from 1 to 2 log10 lower than the expected copy number calculated by RT-qPCR standard curve. The sensitivity of the RT-qPCR and RT-qPCR array assays was found to be similar for two viruses, and better with RT-qPCR than with RT-qPCR array for eighteen viruses by a factor range of from 0.7 to 3.0 log10. Conversely, sensitivity was only 0.30 log10 better with the RT-qPCR array than with conventional RT-qPCR assays for norovirus GIV detection. Finally, the RT-qPCR array and RT-dPCR assays were successfully used together to screen clinical samples and quantify pathogenic viruses. Additionally, this method made it possible to identify co-infection in clinical samples. In conclusion, given the rapidity and potential for large numbers of viral targets, this nanofluidic RT-qPCR assay should have a major impact on human pathogenic virus surveillance and outbreak investigations and is likely to be of benefit

  3. Axiom turkey genotyping array

    Science.gov (United States)

    The Axiom®Turkey Genotyping Array interrogates 643,845 probesets on the array, covering 643,845 SNPs. The array development was led by Dr. Julie Long of the USDA-ARS Beltsville Agricultural Research Center under a public-private partnership with Hendrix Genetics, Aviagen, and Affymetrix. The Turk...

  4. Quantitative (real-time) PCR

    International Nuclear Information System (INIS)

    Denman, S.E.; McSweeney, C.S.

    2005-01-01

    Many nucleic acid-based probe and PCR assays have been developed for the detection tracking of specific microbes within the rumen ecosystem. Conventional PCR assays detect PCR products at the end stage of each PCR reaction, where exponential amplification is no longer being achieved. This approach can result in different end product (amplicon) quantities being generated. In contrast, using quantitative, or real-time PCR, quantification of the amplicon is performed not at the end of the reaction, but rather during exponential amplification, where theoretically each cycle will result in a doubling of product being created. For real-time PCR, the cycle at which fluorescence is deemed to be detectable above the background during the exponential phase is termed the cycle threshold (Ct). The Ct values obtained are then used for quantitation, which will be discussed later

  5. Setup of a PCR laboratory.

    Science.gov (United States)

    Khan, Zaheer

    2011-01-01

    PCR represents an extremely powerful and central molecular biology method. At the heart of its power is the exquisite sensitivity offered: single molecule detection in certain contexts. However, with great power comes great responsibility. Contamination of reagents or test samples with amplifiable material, such as previous reaction products, can be crippling to scientists applying PCR protocols. Prevention of PCR contamination is far and away preferred over eradication. This chapter sets out to offer guidance as to how to use PCR while minimising contamination problems.

  6. Carbon nanotube nanoelectrode arrays

    Science.gov (United States)

    Ren, Zhifeng; Lin, Yuehe; Yantasee, Wassana; Liu, Guodong; Lu, Fang; Tu, Yi

    2008-11-18

    The present invention relates to microelectode arrays (MEAs), and more particularly to carbon nanotube nanoelectrode arrays (CNT-NEAs) for chemical and biological sensing, and methods of use. A nanoelectrode array includes a carbon nanotube material comprising an array of substantially linear carbon nanotubes each having a proximal end and a distal end, the proximal end of the carbon nanotubes are attached to a catalyst substrate material so as to form the array with a pre-determined site density, wherein the carbon nanotubes are aligned with respect to one another within the array; an electrically insulating layer on the surface of the carbon nanotube material, whereby the distal end of the carbon nanotubes extend beyond the electrically insulating layer; a second adhesive electrically insulating layer on the surface of the electrically insulating layer, whereby the distal end of the carbon nanotubes extend beyond the second adhesive electrically insulating layer; and a metal wire attached to the catalyst substrate material.

  7. Phased-array radars

    Science.gov (United States)

    Brookner, E.

    1985-02-01

    The operating principles, technology, and applications of phased-array radars are reviewed and illustrated with diagrams and photographs. Consideration is given to the antenna elements, circuitry for time delays, phase shifters, pulse coding and compression, and hybrid radars combining phased arrays with lenses to alter the beam characteristics. The capabilities and typical hardware of phased arrays are shown using the US military systems COBRA DANE and PAVE PAWS as examples.

  8. Integrated avalanche photodiode arrays

    Science.gov (United States)

    Harmon, Eric S.

    2015-07-07

    The present disclosure includes devices for detecting photons, including avalanche photon detectors, arrays of such detectors, and circuits including such arrays. In some aspects, the detectors and arrays include a virtual beveled edge mesa structure surrounded by resistive material damaged by ion implantation and having side wall profiles that taper inwardly towards the top of the mesa structures, or towards the direction from which the ion implantation occurred. Other aspects are directed to masking and multiple implantation and/or annealing steps. Furthermore, methods for fabricating and using such devices, circuits and arrays are disclosed.

  9. Cover array string reconstruction

    OpenAIRE

    Crochemore, Maxime; S. Iliopoulos, Costas; P. Pissis, Solon; Tischler, German

    2010-01-01

    International audience; A proper factor u of a string y is a cover of y if every letter of y is within some occurrence of u in y. The concept generalises the notion of periods of a string. An integer array C is the minimal-cover (resp. maximal-cover) array of y if C[i] is the minimal (resp. maximal) length of covers of y[0.. i], or zero if no cover exists. In this paper, we present a constructive algorithm checking the validity of an array as a minimal-cover or maximal-cover array of some str...

  10. Storage array reflection considerations

    International Nuclear Information System (INIS)

    Haire, M.J.; Jordan, W.C.; Taylor, R.G.

    1997-01-01

    The assumptions used for reflection conditions of single containers are fairly well established and consistently applied throughout the industry in nuclear criticality safety evaluations. Containers are usually considered to be either fully water reflected (i.e., surrounded by 6 to 12 in. of water) for safety calculations or reflected by 1 in. of water for nominal (structural material and air) conditions. Tables and figures are usually available for performing comparative evaluations of containers under various loading conditions. Reflection considerations used for evaluating the safety of storage arrays of fissile material are not as well established. When evaluating arrays, it has become more common for analysts to use calculations to demonstrate the safety of the array configuration. In performing these calculations, the analyst has considerable freedom concerning the assumptions made for modeling the reflection of the array. Considerations are given for the physical layout of the array with little or no discussion (or demonstration) of what conditions are bounded by the assumed reflection conditions. For example, an array may be generically evaluated by placing it in a corner of a room in which the opposing walls are far away. Typically, it is believed that complete flooding of the room is incredible, so the array is evaluated for various levels of water mist interspersed among array containers. This paper discusses some assumptions that are made regarding storage array reflection

  11. Solar array stepping to minimize array excitation

    Science.gov (United States)

    Bhat, Mahabaleshwar K. P. (Inventor); Liu, Tung Y. (Inventor); Plescia, Carl T. (Inventor)

    1989-01-01

    Mechanical oscillations of a mechanism containing a stepper motor, such as a solar-array powered spacecraft, are reduced and minimized by the execution of step movements in pairs of steps, the period between steps being equal to one-half of the period of torsional oscillation of the mechanism. Each pair of steps is repeated at needed intervals to maintain desired continuous movement of the portion of elements to be moved, such as the solar array of a spacecraft. In order to account for uncertainty as well as slow change in the period of torsional oscillation, a command unit may be provided for varying the interval between steps in a pair.

  12. Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?

    NARCIS (Netherlands)

    Morton, C.O.; White, P.L.; Barnes, R.A.; Klingspor, L.; Cuenca-Estrella, M.; Lagrou, K.; Bretagne, S.; Melchers, W.J.; Mengoli, C.; Caliendo, A.M.; Cogliati, M.; Debets-Ossenkopp, Y.; Gorton, R.; Hagen, F.; Halliday, C.; Hamal, P.; Harvey-Wood, K.; Jaton, K.; Johnson, G.; Kidd, S.; Lengerova, M.; Lass-Florl, C.; Linton, C.; Millon, L.; Morrissey, C.O.; Paholcsek, M.; Talento, A.F.; Ruhnke, M.; Willinger, B.; Donnelly, J.P.; Loeffler, J.

    2017-01-01

    A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help

  13. Focal plane array with modular pixel array components for scalability

    Science.gov (United States)

    Kay, Randolph R; Campbell, David V; Shinde, Subhash L; Rienstra, Jeffrey L; Serkland, Darwin K; Holmes, Michael L

    2014-12-09

    A modular, scalable focal plane array is provided as an array of integrated circuit dice, wherein each die includes a given amount of modular pixel array circuitry. The array of dice effectively multiplies the amount of modular pixel array circuitry to produce a larger pixel array without increasing die size. Desired pixel pitch across the enlarged pixel array is preserved by forming die stacks with each pixel array circuitry die stacked on a separate die that contains the corresponding signal processing circuitry. Techniques for die stack interconnections and die stack placement are implemented to ensure that the desired pixel pitch is preserved across the enlarged pixel array.

  14. Digital PCR: A brief history

    OpenAIRE

    Morley, Alexander A.

    2014-01-01

    Digital PCR for quantification of a target of interest has been independently developed several times, being described in 1990 and 1991 using the term “limiting dilution PCR” and in 1999 using the term “digital PCR”. It came into use in the decade following its first development but its use was cut short by the description of real-time PCR in 1996. However digital PCR has now had a renaissance due to the recent development of new instruments and chemistry which have made it a much simpler and...

  15. Micromirror array for protein micro array fabrication

    Science.gov (United States)

    Lee, Kook-Nyung; Shin, Dong-Sik; Lee, Yoon-Sik; Kim, Yong-Kweon

    2003-05-01

    We have designed, fabricated and characterized a micromirror array for protein microarray fabrication that has a simple structure, and the straightforward fabrication process for the mirror will allow the use of low-cost mirrors in protein pattern applications. The characteristics of an exposure system utilizing the micromirror array have been demonstrated by means of an experiment employing a photoresist that is in general use in the semiconductor industry. The micromirror dimensions were 54 × 54 mum2, with a 30 mum separation between mirrors, and a 5.5 mum gap from the bottom electrode so that the mirror had an approximately 10° deflection angle. The size and separation of the mirror were designed in consideration of the protein pattern size and pitch, in contrast with the commercial Texas Instruments Digital Light Processor, which is utilized in the components of projection display systems. The exposure system combined with the micromirror has been used in the photochemical synthesis of chemical ligands via protein immobilization on a chip. Several photosynthesis experiments for peptide array synthesis have been carried out using the micromirror array. Parallel experiments on photochemical ligand synthesis on a chip can easily be performed in the laboratory using this exposure system.

  16. Triggering the GRANDE array

    International Nuclear Information System (INIS)

    Wilson, C.L.; Bratton, C.B.; Gurr, J.; Kropp, W.; Nelson, M.; Sobel, H.; Svoboda, R.; Yodh, G.; Burnett, T.; Chaloupka, V.; Wilkes, R.J.; Cherry, M.; Ellison, S.B.; Guzik, T.G.; Wefel, J.; Gaidos, J.; Loeffler, F.; Sembroski, G.; Goodman, J.; Haines, T.J.; Kielczewska, D.; Lane, C.; Steinberg, R.; Lieber, M.; Nagle, D.; Potter, M.; Tripp, R.

    1990-01-01

    A brief description of the Gamma Ray And Neutrino Detector Experiment (GRANDE) is presented. The detector elements and electronics are described. The trigger logic for the array is then examined. The triggers for the Gamma Ray and the Neutrino portions of the array are treated separately. (orig.)

  17. Micromachined electrode array

    Science.gov (United States)

    Okandan, Murat; Wessendorf, Kurt O.

    2007-12-11

    An electrode array is disclosed which has applications for neural stimulation and sensing. The electrode array, in certain embodiments, can include a plurality of electrodes each of which is flexibly attached to a common substrate using a plurality of springs to allow the electrodes to move independently. In other embodiments of the electrode array, the electrodes can be fixed to the substrate. The electrode array can be formed from a combination of bulk and surface micromachining, and can include electrode tips having an electroplated metal (e.g. platinum, iridium, gold or titanium) or a metal oxide (e.g. iridium oxide) for biocompatibility. The electrode array can be used to form a part of a neural prosthesis, and is particularly well adapted for use in an implantable retinal prosthesis.

  18. Sensor array signal processing

    CERN Document Server

    Naidu, Prabhakar S

    2009-01-01

    Chapter One: An Overview of Wavefields 1.1 Types of Wavefields and the Governing Equations 1.2 Wavefield in open space 1.3 Wavefield in bounded space 1.4 Stochastic wavefield 1.5 Multipath propagation 1.6 Propagation through random medium 1.7 ExercisesChapter Two: Sensor Array Systems 2.1 Uniform linear array (ULA) 2.2 Planar array 2.3 Distributed sensor array 2.4 Broadband sensor array 2.5 Source and sensor arrays 2.6 Multi-component sensor array2.7 ExercisesChapter Three: Frequency Wavenumber Processing 3.1 Digital filters in the w-k domain 3.2 Mapping of 1D into 2D filters 3.3 Multichannel Wiener filters 3.4 Wiener filters for ULA and UCA 3.5 Predictive noise cancellation 3.6 Exercises Chapter Four: Source Localization: Frequency Wavenumber Spectrum4.1 Frequency wavenumber spectrum 4.2 Beamformation 4.3 Capon's w-k spectrum 4.4 Maximum entropy w-k spectrum 4.5 Doppler-Azimuth Processing4.6 ExercisesChapter Five: Source Localization: Subspace Methods 5.1 Subspace methods (Narrowband) 5.2 Subspace methods (B...

  19. Piezoelectric transducer array microspeaker

    KAUST Repository

    Carreno, Armando Arpys Arevalo

    2016-12-19

    In this paper we present the fabrication and characterization of a piezoelectric micro-speaker. The speaker is an array of micro-machined piezoelectric membranes, fabricated on silicon wafer using advanced micro-machining techniques. Each array contains 2n piezoelectric transducer membranes, where “n” is the bit number. Every element of the array has a circular shape structure. The membrane is made out four layers: 300nm of platinum for the bottom electrode, 250nm or lead zirconate titanate (PZT), a top electrode of 300nm and a structural layer of 50

  20. Introduction to adaptive arrays

    CERN Document Server

    Monzingo, Bob; Haupt, Randy

    2011-01-01

    This second edition is an extensive modernization of the bestselling introduction to the subject of adaptive array sensor systems. With the number of applications of adaptive array sensor systems growing each year, this look at the principles and fundamental techniques that are critical to these systems is more important than ever before. Introduction to Adaptive Arrays, 2nd Edition is organized as a tutorial, taking the reader by the hand and leading them through the maze of jargon that often surrounds this highly technical subject. It is easy to read and easy to follow as fundamental concept

  1. Photonic Crystal Nanocavity Arrays

    National Research Council Canada - National Science Library

    Altug, Hatice; Vuckovic, Jelena

    2006-01-01

    We recently proposed two-dimensional coupled photonic crystal nanocavity arrays as a route to achieve a slow-group velocity of light in all crystal directions, thereby enabling numerous applications...

  2. Flexible retinal electrode array

    Science.gov (United States)

    Okandan, Murat [Albuquerque, NM; Wessendorf, Kurt O [Albuquerque, NM; Christenson, Todd R [Albuquerque, NM

    2006-10-24

    An electrode array which has applications for neural stimulation and sensing. The electrode array can include a large number of electrodes each of which is flexibly attached to a common substrate using a plurality of springs to allow the electrodes to move independently. The electrode array can be formed from a combination of bulk and surface micromachining, with electrode tips that can include an electroplated metal (e.g. platinum, iridium, gold or titanium) or a metal oxide (e.g. iridium oxide) for biocompatibility. The electrode array can be used to form a part of a neural prosthesis, and is particularly well adapted for use in an implantable retinal prosthesis where the electrodes can be tailored to provide a uniform gentle contact pressure with optional sensing of this contact pressure at one or more of the electrodes.

  3. Protein Functionalized Nanodiamond Arrays

    Directory of Open Access Journals (Sweden)

    Liu YL

    2010-01-01

    Full Text Available Abstract Various nanoscale elements are currently being explored for bio-applications, such as in bio-images, bio-detection, and bio-sensors. Among them, nanodiamonds possess remarkable features such as low bio-cytotoxicity, good optical property in fluorescent and Raman spectra, and good photostability for bio-applications. In this work, we devise techniques to position functionalized nanodiamonds on self-assembled monolayer (SAMs arrays adsorbed on silicon and ITO substrates surface using electron beam lithography techniques. The nanodiamond arrays were functionalized with lysozyme to target a certain biomolecule or protein specifically. The optical properties of the nanodiamond-protein complex arrays were characterized by a high throughput confocal microscope. The synthesized nanodiamond-lysozyme complex arrays were found to still retain their functionality in interacting with E. coli.

  4. Expandable LED array interconnect

    Science.gov (United States)

    Yuan, Thomas Cheng-Hsin; Keller, Bernd

    2011-03-01

    A light emitting device that can function as an array element in an expandable array of such devices. The light emitting device comprises a substrate that has a top surface and a plurality of edges. Input and output terminals are mounted to the top surface of the substrate. Both terminals comprise a plurality of contact pads disposed proximate to the edges of the substrate, allowing for easy access to both terminals from multiple edges of the substrate. A lighting element is mounted to the top surface of the substrate. The lighting element is connected between the input and output terminals. The contact pads provide multiple access points to the terminals which allow for greater flexibility in design when the devices are used as array elements in an expandable array.

  5. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  6. Evaluation of Microarray Preprocessing Algorithms Based on Concordance with RT-PCR in Clinical Samples

    DEFF Research Database (Denmark)

    Hansen, Kasper Lage; Szallasi, Zoltan Imre; Eklund, Aron Charles

    2009-01-01

    performed. METHODOLOGY/PRINCIPAL FINDINGS: We used TaqMan RT-PCR arrays as a reference to evaluate the accuracy of expression values from Affymetrix microarrays in two experimental data sets: one comprising 84 genes in 36 colon biopsies, and the other comprising 75 genes in 29 cancer cell lines. We...

  7. Testing of focal plane arrays

    International Nuclear Information System (INIS)

    Merriam, J.D.

    1988-01-01

    Problems associated with the testing of focal plane arrays are briefly examined with reference to the instrumentation and measurement procedures. In particular, the approach and instrumentation used as the Naval Ocean Systems Center is presented. Most of the measurements are made with flooded illumination on the focal plane array. The array is treated as an ensemble of individual pixels, data being taken on each pixel and array averages and standard deviations computed for the entire array. Data maps are generated, showing the pixel data in the proper spatial position on the array and the array statistics

  8. PCR

    African Journals Online (AJOL)

    Administrator

    2006-10-02

    Oct 2, 2006 ... RP-N (15941482) ..... Cloning of the nucleocapsid protein gene of peste des petits ruminants virus: relationship to other Morbillivuruses. J. General Virol. 75:233-237. Diallo A, Barrett T, Barbron M, Shaila MS, Taylor WP ...

  9. pcr

    African Journals Online (AJOL)

    DR. AMINU

    These include DNA cloning for sequencing, DNA based phylogeny, or functional analysis of genes; the diagnosis of hereditary diseases; the identification of genetic finger prints (used in ... Strand separation (denaturing) of the double stranded sample .... study with 30 clinical specimen 18AH and 12VF from. 20 eyes with the ...

  10. Photovoltaic array reliability optimization

    Science.gov (United States)

    Ross, R. G., Jr.

    1982-01-01

    An overview of the photovoltaic array reliability problem is presented, and a high reliability/minimum cost approach to this problem is presented. Design areas covered are cell failure, interconnect fatigue, and electrical insulation breakdown, and three solution strategies are discussed. The first involves controlling component failures in the solar cell (cell cracking, cell interconnects) and at the module level (must be statistically treated). Second, a fault tolerant circuit is designed which reduces array degradation, improves module yield losses, and controls hot-spot heating. Third, cost optimum module replacement strategies are also effective in reducing array degradation. This can be achieved by minimizing the life-cycle energy cost of the photovoltaic system. The integration of these solutions is aimed at reducing the 0.01% failure rate.

  11. Coaxial phased array antenna

    Science.gov (United States)

    Ellis, H., Jr.

    1980-08-01

    A coaxial antenna array for communicating circularly polarized electromagnetic radiation is disclosed. A pair of open ended antenna cavities is coaxially constructed and operates by excitation of linear radiation elements arranged within each of the cavities. A pair of crossed dipole radiation devices is centered within the inner cavity and operated by means of a phase shifting network circuit to transmit as well as receive circularly polarized radiation. Four monopole radiation devices are symmetrically arranged to operate in the outer cavity in phase quadrature by means of the phase shifting network circuit to both transmit and receive circularly polarized electromagnetic radiation. Combined operation of the two antenna cavities with a 180 deg phase differential between the fields related to the two antenna cavities provides a broad beam, relatively wide frequency bandwidth communication capability. Particular embodiments disclosed feature a generally square cavity array as well as a circular cavity array.

  12. Micromachined droplet ejector arrays

    Science.gov (United States)

    Perçin, Gökhan; Yaralioglu, Göksenin G.; Khuri-Yakub, Butrus T.

    2002-12-01

    In this article we present a micromachined flextensional droplet ejector array used to eject liquids. By placing a fluid behind one face of a vibrating circular plate that has an orifice at its center, we achieve continuous ejection of the fluid. We present results of ejection of water and isopropanol. The ejector is harmless to sensitive fluids and can be used to eject fuels, organic polymers, photoresists, low-k dielectrics, adhesives, and chemical and biological samples. Micromachined two-dimensional array flextensional droplet ejectors were realized using planar silicon micromachining techniques. Typical resonant frequency of the micromachined device ranges from 400 kHz to 4.5 MHz. The ejections of water through a 4 μm diameter orifice at 3.45 MHz and a 10 μm diameter orifice at 2.15 MHz were demonstrated by using the developed micromachined two-dimensional array ejectors.

  13. Wire Array Photovoltaics

    Science.gov (United States)

    Turner-Evans, Dan

    Over the past five years, the cost of solar panels has dropped drastically and, in concert, the number of installed modules has risen exponentially. However, solar electricity is still more than twice as expensive as electricity from a natural gas plant. Fortunately, wire array solar cells have emerged as a promising technology for further lowering the cost of solar. Si wire array solar cells are formed with a unique, low cost growth method and use 100 times less material than conventional Si cells. The wires can be embedded in a transparent, flexible polymer to create a free-standing array that can be rolled up for easy installation in a variety of form factors. Furthermore, by incorporating multijunctions into the wire morphology, higher efficiencies can be achieved while taking advantage of the unique defect relaxation pathways afforded by the 3D wire geometry. The work in this thesis shepherded Si wires from undoped arrays to flexible, functional large area devices and laid the groundwork for multijunction wire array cells. Fabrication techniques were developed to turn intrinsic Si wires into full p-n junctions and the wires were passivated with a-Si:H and a-SiNx:H. Single wire devices yielded open circuit voltages of 600 mV and efficiencies of 9%. The arrays were then embedded in a polymer and contacted with a transparent, flexible, Ni nanoparticle and Ag nanowire top contact. The contact connected >99% of the wires in parallel and yielded flexible, substrate free solar cells featuring hundreds of thousands of wires. Building on the success of the Si wire arrays, GaP was epitaxially grown on the material to create heterostructures for photoelectrochemistry. These cells were limited by low absorption in the GaP due to its indirect bandgap, and poor current collection due to a diffusion length of only 80 nm. However, GaAsP on SiGe offers a superior combination of materials, and wire architectures based on these semiconductors were investigated for multijunction

  14. A review of array radars

    Science.gov (United States)

    Brookner, E.

    1981-10-01

    Achievements in the area of array radars are illustrated by such activities as the operational deployment of the large high-power, high-range-resolution Cobra Dane; the operational deployment of two all-solid-state high-power, large UHF Pave Paws radars; and the development of the SAM multifunction Patriot radar. This paper reviews the following topics: array radars steered in azimuth and elevation by phase shifting (phase-phase steered arrays); arrays steered + or - 60 deg, limited scan arrays, hemispherical coverage, and omnidirectional coverage arrays; array radars steering electronically in only one dimension, either by frequency or by phase steering; and array radar antennas which use no electronic scanning but instead use array antennas for achieving low antenna sidelobes.

  15. Array Theory and Nial

    DEFF Research Database (Denmark)

    Falster, Peter; Jenkins, Michael

    1999-01-01

    This report is the result of collaboration between the authors during the first 8 months of 1999 when M. Jenkins was visiting professor at DTU. The report documents the development of a tool for the investigation of array theory concepts and in particular presents various approaches to choose...

  16. Two-temperature PCR for Microfluidics

    KAUST Repository

    Kodzius, Rimantas

    2010-05-01

    Since its invention in 1983, polymerase chain reaction (PCR) has been the method of choice for DNA amplification. Successful PCR depends on the optimization of several parameters, which is a cumbersome task due to the many variables (conditions and compon

  17. Radar techniques using array antennas

    CERN Document Server

    Wirth, Wulf-Dieter

    2013-01-01

    Radar Techniques Using Array Antennas is a thorough introduction to the possibilities of radar technology based on electronic steerable and active array antennas. Topics covered include array signal processing, array calibration, adaptive digital beamforming, adaptive monopulse, superresolution, pulse compression, sequential detection, target detection with long pulse series, space-time adaptive processing (STAP), moving target detection using synthetic aperture radar (SAR), target imaging, energy management and system parameter relations. The discussed methods are confirmed by simulation stud

  18. DNA Sequence Signatures for Rapid Detection of Six Target Bacterial Pathogens Using PCR Assays

    Directory of Open Access Journals (Sweden)

    Kenjiro Nagamine

    2015-01-01

    Full Text Available Using Streptococcus pyogenes as a model, we previously established a stepwise computational workflow to effectively identify species-specific DNA signatures that could be used as PCR primer sets to detect target bacteria with high specificity and sensitivity. In this study, we extended the workflow for the rapid development of PCR assays targeting Enterococcus faecalis, Enterococcus faecium, Clostridium perfringens, Clostridium difficile, Clostridium tetani , and Staphylococcus aureus , which are of safety concern for human tissue intended for transplantation. Twenty-one primer sets that had sensitivity of detecting 5–50 fg DNA from target bacteria with high specificity were selected. These selected primer sets can be used in a PCR array for detecting target bacteria with high sensitivity and specificity. The workflow could be widely applicable for the rapid development of PCR-based assays for a wide range of target bacteria, including those of biothreat agents.

  19. Pathogen Causing Disease of Diagnosis PCR Tecnology

    OpenAIRE

    SEVİNDİK, Emre; KIR, A. Çağrı; BAŞKEMER, Kadir; UZUN, Veysel

    2013-01-01

    Polimerase chain reaction (PCR) with which, the development of recombinant DNA tecnology, a technique commonly used in field of moleculer biology and genetic. Duplication of the target DNA is provided with this technique without the need for cloning. Some fungus species, bacteria, viruses constitutent an important group of pathogenicity in human, animals and plants. There are routinely applied types of PCR in the detection of pathogens infections diseases. These Nested- PCR, Real- Time PCR, M...

  20. Timed arrays wideband and time varying antenna arrays

    CERN Document Server

    Haupt, Randy L

    2015-01-01

    Introduces timed arrays and design approaches to meet the new high performance standards The author concentrates on any aspect of an antenna array that must be viewed from a time perspective. The first chapters briefly introduce antenna arrays and explain the difference between phased and timed arrays. Since timed arrays are designed for realistic time-varying signals and scenarios, the book also reviews wideband signals, baseband and passband RF signals, polarization and signal bandwidth. Other topics covered include time domain, mutual coupling, wideband elements, and dispersion. The auth

  1. Seismometer array station processors

    International Nuclear Information System (INIS)

    Key, F.A.; Lea, T.G.; Douglas, A.

    1977-01-01

    A description is given of the design, construction and initial testing of two types of Seismometer Array Station Processor (SASP), one to work with data stored on magnetic tape in analogue form, the other with data in digital form. The purpose of a SASP is to detect the short period P waves recorded by a UK-type array of 20 seismometers and to edit these on to a a digital library tape or disc. The edited data are then processed to obtain a rough location for the source and to produce seismograms (after optimum processing) for analysis by a seismologist. SASPs are an important component in the scheme for monitoring underground explosions advocated by the UK in the Conference of the Committee on Disarmament. With digital input a SASP can operate at 30 times real time using a linear detection process and at 20 times real time using the log detector of Weichert. Although the log detector is slower, it has the advantage over the linear detector that signals with lower signal-to-noise ratio can be detected and spurious large amplitudes are less likely to produce a detection. It is recommended, therefore, that where possible array data should be recorded in digital form for input to a SASP and that the log detector of Weichert be used. Trial runs show that a SASP is capable of detecting signals down to signal-to-noise ratios of about two with very few false detections, and at mid-continental array sites it should be capable of detecting most, if not all, the signals with magnitude above msub(b) 4.5; the UK argues that, given a suitable network, it is realistic to hope that sources of this magnitude and above can be detected and identified by seismological means alone. (author)

  2. Solar collector array

    Science.gov (United States)

    Hall, John Champlin; Martins, Guy Lawrence

    2015-09-06

    A method and apparatus for efficient manufacture, assembly and production of solar energy. In one aspect, the apparatus may include a number of modular solar receiver assemblies that may be separately manufactured, assembled and individually inserted into a solar collector array housing shaped to receive a plurality of solar receivers. The housing may include optical elements for focusing light onto the individual receivers, and a circuit for electrically connecting the solar receivers.

  3. Array processor architecture

    Science.gov (United States)

    Barnes, George H. (Inventor); Lundstrom, Stephen F. (Inventor); Shafer, Philip E. (Inventor)

    1983-01-01

    A high speed parallel array data processing architecture fashioned under a computational envelope approach includes a data base memory for secondary storage of programs and data, and a plurality of memory modules interconnected to a plurality of processing modules by a connection network of the Omega gender. Programs and data are fed from the data base memory to the plurality of memory modules and from hence the programs are fed through the connection network to the array of processors (one copy of each program for each processor). Execution of the programs occur with the processors operating normally quite independently of each other in a multiprocessing fashion. For data dependent operations and other suitable operations, all processors are instructed to finish one given task or program branch before all are instructed to proceed in parallel processing fashion on the next instruction. Even when functioning in the parallel processing mode however, the processors are not locked-step but execute their own copy of the program individually unless or until another overall processor array synchronization instruction is issued.

  4. Spaceborne Processor Array

    Science.gov (United States)

    Chow, Edward T.; Schatzel, Donald V.; Whitaker, William D.; Sterling, Thomas

    2008-01-01

    A Spaceborne Processor Array in Multifunctional Structure (SPAMS) can lower the total mass of the electronic and structural overhead of spacecraft, resulting in reduced launch costs, while increasing the science return through dynamic onboard computing. SPAMS integrates the multifunctional structure (MFS) and the Gilgamesh Memory, Intelligence, and Network Device (MIND) multi-core in-memory computer architecture into a single-system super-architecture. This transforms every inch of a spacecraft into a sharable, interconnected, smart computing element to increase computing performance while simultaneously reducing mass. The MIND in-memory architecture provides a foundation for high-performance, low-power, and fault-tolerant computing. The MIND chip has an internal structure that includes memory, processing, and communication functionality. The Gilgamesh is a scalable system comprising multiple MIND chips interconnected to operate as a single, tightly coupled, parallel computer. The array of MIND components shares a global, virtual name space for program variables and tasks that are allocated at run time to the distributed physical memory and processing resources. Individual processor- memory nodes can be activated or powered down at run time to provide active power management and to configure around faults. A SPAMS system is comprised of a distributed Gilgamesh array built into MFS, interfaces into instrument and communication subsystems, a mass storage interface, and a radiation-hardened flight computer.

  5. Lectin-Array Blotting.

    Science.gov (United States)

    Pazos, Raquel; Echevarria, Juan; Hernandez, Alvaro; Reichardt, Niels-Christian

    2017-09-01

    Aberrant protein glycosylation is a hallmark of cancer, infectious diseases, and autoimmune or neurodegenerative disorders. Unlocking the potential of glycans as disease markers will require rapid and unbiased glycoproteomics methods for glycan biomarker discovery. The present method is a facile and rapid protocol for qualitative analysis of protein glycosylation in complex biological mixtures. While traditional lectin arrays only provide an average signal for the glycans in the mixture, which is usually dominated by the most abundant proteins, our method provides individual lectin binding profiles for all proteins separated in the gel electrophoresis step. Proteins do not have to be excised from the gel for subsequent analysis via the lectin array but are transferred by contact diffusion from the gel to a glass slide presenting multiple copies of printed lectin arrays. Fluorescently marked glycoproteins are trapped by the printed lectins via specific carbohydrate-lectin interactions and after a washing step their binding profile with up to 20 lectin probes is analyzed with a fluorescent scanner. The method produces the equivalent of 20 lectin blots in a single experiment, giving detailed insight into the binding epitopes present in the fractionated proteins. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  6. Principles and technical aspects of PCR amplification

    National Research Council Canada - National Science Library

    Pelt-Verkuil, Elizabeth van; Belkum, Alex van; Hays, John P

    2008-01-01

    ... to illustrate any particularly important concepts or comments. Indeed, all commercial PCR biotechnology companies offer information about their products on internet sites and in online technical manuals. These online resources will be invaluable for any readers requiring more detailed PCR protocols. The authors have provided references for many PCR co...

  7. The PCR revolution: basic technologies and applications

    National Research Council Canada - National Science Library

    Bustin, Stephen A

    2010-01-01

    ... by leading authorities on the many applications of PCR and how this technology has revolutionized their respective areas of interest. This book conveys the ways in which PCR has overcome many obstacles in life science and clinical research and also charts the PCR's development from time-consuming, low throughput, nonquantitative proced...

  8. Human minisatellite alleles detectable only after PCR amplification.

    Science.gov (United States)

    Armour, J A; Crosier, M; Jeffreys, A J

    1992-01-01

    We present evidence that a proportion of alleles at two human minisatellite loci is undetected by standard Southern blot hybridization. In each case the missing allele(s) can be identified after PCR amplification and correspond to tandem arrays too short to detect by hybridization. At one locus, there is only one undetected allele (population frequency 0.3), which contains just three repeat units. At the second locus, there are at least five undetected alleles (total population frequency 0.9) containing 60-120 repeats; they are not detected because these tandem repeats give very poor signals when used as a probe in standard Southern blot hybridization, and also cross-hybridize with other sequences in the genome. Under these circumstances only signals from the longest tandemly repeated alleles are detectable above the nonspecific background. The structures of these loci have been compared in human and primate DNA, and at one locus the short human allele containing three repeat units is shown to be an intermediate state in the expansion of a monomeric precursor allele in primates to high copy number in the longer human arrays. We discuss the implications of such loci for studies of human populations, minisatellite isolation by cloning, and the evolution of highly variable tandem arrays.

  9. Real-time PCR in Food Science: PCR Diagnostics.

    Science.gov (United States)

    Rodriguez-Lazaro, David; Cook, Nigel; Hernandez, Marta

    2013-01-01

    A principal consumer demand is a guarantee of the safety and quality of food. The presence of foodborne pathogens and their potential hazard, the use of genetically modified organisms (GMOs) in food production, and the correct labelling in foods suitable for vegetarians are among the subjects where society demands total transparency. The application of controls within the quality assessment programmes of the food industry is a way to satisfy these demands, and is necessary to ensure efficient analytical methodologies are possessed and correctly applied by the Food Sector. The use of real-time PCR has become a promising alternative approach in food diagnostics. It possesses a number of advantages over conventional culturing approaches, including rapidity, excellent analytical sensitivity and selectivity, and potential for quantification. However, the use of expensive equipment and reagents, the need for qualified personnel, and the lack of standardized protocols are impairing its practical implementation for food monitoring and control.

  10. [Progress in digital PCR technology and application].

    Science.gov (United States)

    Lin, Jiaqi; Su, Guocheng; Su, Wenjin; Zhou, Changyi

    2017-02-25

    Digital PCR is an emerging analysis technology for absolute quantification after realtime-PCR. Through digital PCR, single DNA molecules are distributed into isolated reactions, and the product with fluorescence signal can be detected and analyzed after amplification. With the advantages of higher sensitivity and accuracy, digital PCR, independent of a standard curve, is developing rapidly and applied widely to the next generation sequencing and detection fields, such as gene mutation, copy number variation, microorganism, and genetically modified food. In this article, we reviewed the quantitative method and research progress of digital PCR technology in the main application fields.

  11. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  12. Radiation from waveguide arrays

    International Nuclear Information System (INIS)

    Brambilla, M.

    1977-07-01

    The theory of phased waveguide arrays (the 'Grill'), developed in view of Lower Hybrid Heating of toroidal plasmas, is applied to two simple cases, in order to gain insight on field distributions and power flow. First, the far-field radiation pattern of the Grill towards an empty half-space is evaluated. Next, the excitation of a passive waveguide by a Grill mounted in a T configuration is considered. These results constitue two examples of exact solutions of Maxwell's equations in relatively complex geometry

  13. Molecular logic gate arrays.

    Science.gov (United States)

    de Silva, A Prasanna

    2011-03-01

    Chemists are now able to emulate the ideas and instruments of mathematics and computer science with molecules. The integration of molecular logic gates into small arrays has been a growth area during the last few years. The design principles underlying a collection of these cases are examined. Some of these computing molecules are applicable in medical- and biotechnologies. Cases of blood diagnostics, 'lab-on-a-molecule' systems, and molecular computational identification of small objects are included. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. The Long Wavelength Array

    Science.gov (United States)

    Pihlström, Ylva

    The Long Wavelength Array (LWA) will be a new, open-skies, user-oriented aperture synthesis instrument dedicated to explore frequencies between 20 and 80 MHz. The LWA will provide high temporal (millisecond or better) and high spatial resolution (arcsecond) and mJy-level sensitivity. The LWA key science areas include acceleration, propagation, and turbulence in the ISM; the high-redshift Universe; planetary, solar and space science; and the transient universe at radio wavelengths. In addition, key goals of the LWA are as a training ground for the next generation of radio astronomers and to re-invigorate radio astronomy in the US at the university level. The LWA will be operated by the University of New Mexico on behalf of the South West Consortium (SWC), thereby providing opportunities for students within the fields of astronomy, computer science and electrical engineering. Currently, in its first year of construction funding, the LWA team is now bringing up the first station near the Very Large Array site in the southwest US.

  15. Selecting Sums in Arrays

    DEFF Research Database (Denmark)

    Brodal, Gerth Stølting; Jørgensen, Allan Grønlund

    2008-01-01

    In an array of n numbers each of the \\binomn2+nUnknown control sequence '\\binom' contiguous subarrays define a sum. In this paper we focus on algorithms for selecting and reporting maximal sums from an array of numbers. First, we consider the problem of reporting k subarrays inducing the k largest...... sums among all subarrays of length at least l and at most u. For this problem we design an optimal O(n + k) time algorithm. Secondly, we consider the problem of selecting a subarray storing the k’th largest sum. For this problem we prove a time bound of Θ(n · max {1,log(k/n)}) by describing...... an algorithm with this running time and by proving a matching lower bound. Finally, we combine the ideas and obtain an O(n· max {1,log(k/n)}) time algorithm that selects a subarray storing the k’th largest sum among all subarrays of length at least l and at most u....

  16. Electromagnetically Clean Solar Arrays

    Science.gov (United States)

    Stem, Theodore G.; Kenniston, Anthony E.

    2008-01-01

    The term 'electromagnetically clean solar array' ('EMCSA') refers to a panel that contains a planar array of solar photovoltaic cells and that, in comparison with a functionally equivalent solar-array panel of a type heretofore used on spacecraft, (1) exhibits less electromagnetic interferences to and from other nearby electrical and electronic equipment and (2) can be manufactured at lower cost. The reduction of electromagnetic interferences is effected through a combination of (1) electrically conductive, electrically grounded shielding and (2) reduction of areas of current loops (in order to reduce magnetic moments). The reduction of cost is effected by designing the array to be fabricated as a more nearly unitary structure, using fewer components and fewer process steps. Although EMCSAs were conceived primarily for use on spacecraft they are also potentially advantageous for terrestrial applications in which there are requirements to limit electromagnetic interference. In a conventional solar panel of the type meant to be supplanted by an EMCSA panel, the wiring is normally located on the back side, separated from the cells, thereby giving rise to current loops having significant areas and, consequently, significant magnetic moments. Current-loop geometries are chosen in an effort to balance opposing magnetic moments to limit far-0field magnetic interactions, but the relatively large distances separating current loops makes full cancellation of magnetic fields problematic. The panel is assembled from bare photovoltaic cells by means of multiple sensitive process steps that contribute significantly to cost, especially if electomagnetic cleanliness is desired. The steps include applying a cover glass and electrical-interconnect-cell (CIC) sub-assemble, connecting the CIC subassemblies into strings of series-connected cells, laying down and adhesively bonding the strings onto a panel structure that has been made in a separate multi-step process, and mounting the

  17. Application of reverse transcription-PCR and real-time PCR in nanotoxicity research.

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2012-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq(®) DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix.

  18. Enzootic bovine leukosis real time PCR

    OpenAIRE

    Dias, Natanael Lamas; Fonseca Júnior, Antônio Augusto; Rodrigues, Daniel Sobreira; Camargos, Marcelo Fernandes

    2012-01-01

    O objetivo deste trabalho foi realizar a validação de uma reação em cadeia da polimerase em tempo real com o sistema Plexor® (qPCR) para o diagnóstico da Leucose Enzoótica Bovina (LEB), por meio da comparação com testes de diagnóstico recomendados pela Organização Mundial de Saúde Animal (OIE). A qPCR foi comparada com duas outras técnicas: a PCR nested (nPCR) e a imunodifusão em gel de ágar (IDGA). Das 82 amostras analisadas pela qPCR e nPCR, 79 apresentaram resultados concordantes, sendo a ...

  19. Contaminations Occurring in Fungal PCR Assays

    Science.gov (United States)

    Loeffler, Juergen; Hebart, Holger; Bialek, Ralf; Hagmeyer, Lars; Schmidt, Diethard; Serey, Francois-Prâseth; Hartmann, Matthias; Eucker, Jan; Einsele, Hermann

    1999-01-01

    Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified as Aspergillus fumigatus, Saccharomyces cerevisiae, and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10× PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken. PMID:10074553

  20. Terahertz superconducting plasmonic hole array

    OpenAIRE

    Tian, Zhen; Singh, Ranjan; Han, Jiaguang; Gu, Jianqiang; Xing, Qirong; Zhang, Weili

    2010-01-01

    We demonstrate thermally tunable superconductor hole array with active control over their resonant transmission induced by surface plasmon polaritons . The array was lithographically fabricated on high temperature YBCO superconductor and characterized by terahertz-time domain spectroscopy. We observe a clear transition from the virtual excitation of the surface plasmon mode to the real surface plasmon mode. The highly tunable superconducting plasmonic hole arrays may have promising applicatio...

  1. Combinatorial aspects of covering arrays

    Directory of Open Access Journals (Sweden)

    Charles J. Colbourn

    2004-11-01

    Full Text Available Covering arrays generalize orthogonal arrays by requiring that t -tuples be covered, but not requiring that the appearance of t -tuples be balanced.Their uses in screening experiments has found application in software testing, hardware testing, and a variety of fields in which interactions among factors are to be identified. Here a combinatorial view of covering arrays is adopted, encompassing basic bounds, direct constructions, recursive constructions, algorithmic methods, and applications.

  2. Efficient array design for sonotherapy

    International Nuclear Information System (INIS)

    Stephens, Douglas N; Kruse, Dustin E; Ferrara, Katherine W; Ergun, Arif S; Barnes, Stephen; Lu, X Ming

    2008-01-01

    New linear multi-row, multi-frequency arrays have been designed, constructed and tested as fully operational ultrasound probes to produce confocal imaging and therapeutic acoustic intensities with a standard commercial ultrasound imaging system. The triple-array probes and imaging system produce high quality B-mode images with a center row imaging array at 5.3 MHz and sufficient acoustic power with dual therapeutic arrays to produce mild hyperthermia at 1.54 MHz. The therapeutic array pair in the first probe design (termed G3) utilizes a high bandwidth and peak pressure, suitable for mechanical therapies. The second multi-array design (termed G4) has a redesigned therapeutic array pair which is optimized for a high time-averaged power output suitable for mild hyperthermia applications. The 'thermal therapy' design produces more than 4 W of acoustic power from the low-frequency arrays with only a 10.5 deg. C internal rise in temperature after 100 s of continuous use with an unmodified conventional imaging system or substantially longer operation at lower acoustic power. The low-frequency arrays in both probe designs were examined and contrasted for real power transfer efficiency with a KLM model which includes all lossy contributions in the power delivery path from system transmitters to the tissue load. Laboratory verification was successfully performed for the KLM-derived estimates of transducer parallel model acoustic resistance and dissipation resistance, which are the critical design factors for acoustic power output and undesired internal heating, respectively

  3. Compact dynamic microfluidic iris array

    Science.gov (United States)

    Kimmle, Christina; Doering, Christoph; Steuer, Anna; Fouckhardt, Henning

    2011-09-01

    A dynamic microfluidic iris is realized. Light attenuation is achieved by absorption of an opaque liquid (e.g. black ink). The adjustment of the iris diameter is achieved by fluid displacement via a transparent elastomer (silicone) half-sphere. This silicone calotte is hydraulically pressed against a polymethylmethacrylate (PMMA) substrate as the bottom window, such that the opaque liquid is squeezed away, this way opening the iris. With this approach a dynamic range of more than 60 dB can be achieved with response times in the ms to s regime. The design allows the realization of a single iris as well as an iris array. So far the master for the molded silicone structure was fabricated by precision mechanics. The aperture diameter was changed continuously from 0 to 8 mm for a single iris and 0 to 4 mm in case of a 3 x 3 iris array. Moreover, an iris array was combined with a PMMA lens array into a compact module, the distance of both arrays equaling the focal length of the lenses. This way e.g. spatial frequency filter arrays can be realized. The possibility to extend the iris array concept to an array with many elements is demonstrated. Such arrays could be applied e.g. in light-field cameras.

  4. Nanoelectrode array for electrochemical analysis

    Energy Technology Data Exchange (ETDEWEB)

    Yelton, William G [Sandia Park, NM; Siegal, Michael P [Albuquerque, NM

    2009-12-01

    A nanoelectrode array comprises a plurality of nanoelectrodes wherein the geometric dimensions of the electrode controls the electrochemical response, and the current density is independent of time. By combining a massive array of nanoelectrodes in parallel, the current signal can be amplified while still retaining the beneficial geometric advantages of nanoelectrodes. Such nanoelectrode arrays can be used in a sensor system for rapid, non-contaminating field analysis. For example, an array of suitably functionalized nanoelectrodes can be incorporated into a small, integrated sensor system that can identify many species rapidly and simultaneously under field conditions in high-resistivity water, without the need for chemical addition to increase conductivity.

  5. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Directory of Open Access Journals (Sweden)

    Livia Schrick

    2016-01-01

    Full Text Available PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  6. Absolute quantification by droplet digital PCR versus analog real-time PCR

    Science.gov (United States)

    Hindson, Christopher M; Chevillet, John R; Briggs, Hilary A; Gallichotte, Emily N; Ruf, Ingrid K; Hindson, Benjamin J; Vessella, Robert L; Tewari, Muneesh

    2014-01-01

    Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased by 37–86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer. PMID:23995387

  7. Printed glycan array

    DEFF Research Database (Denmark)

    Shilova, Nadezhda; Navakouski, Maxim; Khasbiullina, Nailya

    2012-01-01

    Using printed glycan array (PGA) we compared the results of antibody profiling in undiluted, moderately (1:15) and highly (1:100) diluted human blood serum. Undiluted serum is suitable for studying blood as a tissue in its native state, whereas to study the serum of newborns or small animals one...... usually has to dilute the starting material in order to have sufficient volume for PGA experimentation. The PGA used in this study allows for the use of whole serum without modifications to the protocol, and the background is surprisingly low. Antibodies profiles observed in undiluted serum versus 1......:15 dilution were similar, with only a limited number of new signals identified in the undiluted serum. However, unexpected irregularities were found when IgG and IgM are measured separately, namely, at a 1:15 dilution more intensive IgG signals for many glycans are observed. We believe that in conditions...

  8. Scintillator detector array

    International Nuclear Information System (INIS)

    Cusano, D.A.; Dibianca, F.A.

    1981-01-01

    This patent application relates to a scintillator detector array for use in computerized tomography and comprises a housing including a plurality of chambers, the said housing having a front wall transmissive to x-rays and side walls opaque to x-rays, such as of tungsten and tantalum, a liquid scintillation medium including a soluble fluor, the solvent for the fluor being disposed in the chambers. The solvent comprises either an intrinsically high Z solvent or a solvent which has dissolved therein a high Z compound e.g. iodo or bromonaphthalene; or toluene, xylene or trimethylbenzene with a lead or tin alkyl dissolved therein. Also disposed about the chambers are a plurality of photoelectric devices. (author)

  9. Field effect sensors for PCR applications

    Science.gov (United States)

    Taing, Meng-Houit; Sweatman, Denis R.

    2004-03-01

    The use of field effect sensors for biological and chemical sensing is widely employed due to its ability to make detections based on charge and surface potential. Because proteins and DNA almost always carry a charge [1], silicon can be used to micro fabricate such a sensor. The EIS structure (Electrolyte on Insulator on Silicon) provides a novel, label-free and simple to fabricate way to make a field effect DNA detection sensor. The sensor responds to fluctuating capacitance caused by a depletion layer thickness change at the surface of the silicon substrate through DNA adsorption onto the dielectric oxide/PLL (Poly-L-Lysine) surface. As DNA molecules diffuse to the sensor surface, they are bound to their complimentary capture probes deposited on the surface. The negative charge exhibited by the DNA forces negative charge carriers in the substrate to move away from the surface. This causes an n-type depletion layer substrate to thicken and a p-type to thin. The depletion layer thickness can be measured by its capacitance using an LCR meter. This experiment is conducted using the ConVolt (constant voltage) approach. Nucleic acids are amplified by an on chip PCR (Polymerase Chain Reaction) system and then fed into the sensor. The low ionic solution strength will ensure that counter-ions do not affect the sensor measurements. The sensor surface contains capture probes that bind to the pathogen. The types of pathogens we"ll be detecting include salmonella, campylobacter and E.Coli DNA. They are held onto the sensor surface by the positively charged Poly-L-Lysine layer. The electrolyte is biased through a pseudo-reference electrode. Pseudo reference electrodes are usually made from metals such as Platinum or Silver. The problem associated with "floating" biasing electrodes is they cannot provide stable biasing potentials [2]. They drift due to surface charging effects and trapped charges on the surface. To eliminate this, a differential system consisting of 2 sensors

  10. Cascading Constrained 2-D Arrays using Periodic Merging Arrays

    DEFF Research Database (Denmark)

    Forchhammer, Søren; Laursen, Torben Vaarby

    2003-01-01

    We consider a method for designing 2-D constrained codes by cascading finite width arrays using predefined finite width periodic merging arrays. This provides a constructive lower bound on the capacity of the 2-D constrained code. Examples include symmetric RLL and density constrained codes...

  11. Report for Detection of Biothreat Agents and Environmental Samples using the LLNL Virulence Array for DHS

    Energy Technology Data Exchange (ETDEWEB)

    Jaing, Crystal [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Gardner, Shea [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); McLoughlin, Kevin [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Thissen, James [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Jackson, Paul [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2011-04-18

    The objective of this project is to provide DHS a comprehensive evaluation of the current genomic technologies including genotyping, Taqman PCR, multiple locus variable tandem repeat analysis (MLVA), microarray and high-throughput DNA sequencing in the analysis of biothreat agents from complex environmental samples. This report focuses on the design, testing and results of samples on the Virulence Array.

  12. Avoiding cross hybridization by choosing nonredundant targets on cDNA arrays

    DEFF Research Database (Denmark)

    Nielsen, Henrik Bjørn; Knudsen, Steen

    2002-01-01

    PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end....

  13. Rice8987 Array: PCR check - RMOS | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available There are independent 8979 cDNA clones that were selected from about 40,000 cDNA clones isolated in the first research... stage of the rice genome research project. Those 8979 cDNA clones were

  14. Gene array and real time PCR analysis of the adrenal sensitivity to adrenocorticotropic hormone in pig

    Directory of Open Access Journals (Sweden)

    SanCristobal Magali

    2008-02-01

    Full Text Available Abstract Background Variability in hypothalamic-pituitary-adrenal (HPA axis activity has been shown to be influenced by genetic factors and related to great metabolic differences such as obesity. The aim of this study was to investigate molecular bases of genetic variability of the adrenal sensitivity to ACTH, a major source of variability, in Meishan (MS and Large White (LW pigs, MS being reported to exhibit higher basal cortisol levels, response to ACTH and fatness than LW. A pig cDNA microarray was used to identify changes in gene expression in basal conditions and in response to ACTH stimulation. Results Genotype and/or ACTH affected the expression of 211 genes related to transcription, cell growth/maintenance, signal transduction, cell structure/adhesion/extra cellular matrix and protein kinase/phosphatase activity. No change in the expression of known key regulator proteins of the ACTH signaling pathway or of steroidogenic enzymes was found. However, Mdh2, Sdha, Suclg2, genes involved in the tricarboxylic acid (TCA pathway, were over-expressed in MS pigs. Higher TCA cycle activity in MS than in LW may thus result in higher steroidogenic activity and thus explain the typically higher cortisol levels in MS compared to LW. Moreover, up-regulation of Star and Ldlr genes in MS and/or in response to ACTH suggest that differences in the adrenal function between MS and LW may also involve mechanisms requisite for cholesterol supply to steroidogenesis. Conclusion The present study provides new potential candidate genes to explain genetic variations in the adrenal sensitivity to ACTH and better understand relationship between HPA axis activity and obesity.

  15. Denaturation strategies for detection of double stranded PCR products on GMR magnetic biosensor array

    DEFF Research Database (Denmark)

    Rizzi, Giovanni; Lee, Jung-Rok; Guldberg, Per

    2017-01-01

    Microarrays and other surface-based nucleic acid detection schemes rely on the hybridization of the target to surface-bound detection probes. We present the first comparison of two strategies to detect DNA using a giant magnetoresistive (GMR) biosensor platform starting from an initially double...

  16. The OncoArray Consortium

    DEFF Research Database (Denmark)

    Amos, Christopher I; Dennis, Joe; Wang, Zhaoming

    2017-01-01

    BACKGROUND: Common cancers develop through a multistep process often including inherited susceptibility. Collaboration among multiple institutions, and funding from multiple sources, has allowed the development of an inexpensive genotyping microarray, the OncoArray. The array includes a genome-wi...

  17. Submillimeter heterodyne arrays for APEX

    NARCIS (Netherlands)

    Güsten, R.; Baryshev, A.; Bell, A.; Belloche, A.; Graf, U.; Hafok, H.; Heyminck, S.; Hochgürtel, S.; Honingh, C. E.; Jacobs, K.; Kasemann, C.; Klein, B.; Klein, T.; Korn, A.; Krämer, I.; Leinz, C.; Lundgren, A.; Menten, K. M.; Meyer, K.; Muders, D.; Pacek, F.; Rabanus, D.; Schäfer, F.; Schilke, P.; Schneider, G.; Stutzki, J.; Wieching, G.; Wunsch, A.; Wyrowski, F.

    2008-01-01

    We report on developments of submillimeter heterodyne arrays for high resolution spectroscopy with APEX. Shortly, we will operate state-of-the-art instruments in all major atmospheric windows accessible from Llano de Chajnantor. CHAMP+, a dual-color 2×7 element heterodyne array for operation in the

  18. Subtyping of swine influenza viruses using a high-throughput real time PCR platform

    DEFF Research Database (Denmark)

    Goecke, Nicole Bakkegård; Krog, Jesper Schak; Hjulsager, Charlotte Kristiane

    for the internal genes of IAV were validated and optimised to run under identical reaction conditions and assembled on a dynamic array chip (Fluidigm). Results. The sensitivity and specificity of the chip was assessed by testing cell culture isolates and field samples with known subtypes (based on sequencing...... test and subsequent subtyping is performed by real time RT-PCR (RT-qPCR) but several assays are needed to cover the wide range of circulating subtypes which is expensive,resource and time demanding. To mitigate these restrictions the high-throughput qPCR platform BioMark (Fluidigm) has been explored...... services with reduced price and turnover time which will facilitate choice of vaccines and by that lead to reduction of antibiotic used....

  19. PCR protocols: current methods and applications

    National Research Council Canada - National Science Library

    White, Bruce Alan

    1993-01-01

    ..." between "small" and "big" labs, since its use makes certain projects, especially those related to molecular cloning, now far more feasible for the small lab with a modest budget. This new volume on PCR Protocols does not attempt the impossible task of representing all PCR-based protocols. Rather, it presents a range of protocols, both analytical ...

  20. [E-MTAB-587] PCR_artifacts

    NARCIS (Netherlands)

    Muino Acuna, J.M.

    2011-01-01

    WARNING: This library was yield low amount of material and it was over-amplified by PCR. This libraries are used study the robustness of several statitical methods against PCR artifacts. ChIP experiments were performed on Arabidopsis wildtype inflorescences using an antibody raised against a

  1. PCR specific for Actinobacillus pleuropneumoniae serotype 3

    DEFF Research Database (Denmark)

    Zhou, L.; Jones, S.C.P.; Angen, Øystein

    2008-01-01

    , but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266 strains...

  2. Testing for Genetically Modified Foods Using PCR

    Science.gov (United States)

    Taylor, Ann; Sajan, Samin

    2005-01-01

    The polymerase chain reaction (PCR) is a Nobel Prize-winning technique that amplifies a specific segment of DNA and is commonly used to test for the presence of genetic modifications. Students use PCR to test corn meal and corn-muffin mixes for the presence of a promoter commonly used in genetically modified foods, the cauliflower mosaic virus 35S…

  3. (PCR) and loop-mediated isothermal amplification

    African Journals Online (AJOL)

    SAM

    2014-03-26

    Mar 26, 2014 ... better alternative for PCR, even in low technology laboratories. In addition, these findings revealed that the possibility of fatal fusariosis due to F. solani is not so rare in HIV positive patients. Key words: Fusarium solani, loop-mediated isothermal amplification (LAMP), HIV, polymerase chain reaction. (PCR).

  4. Validation of RNAi by real time PCR

    DEFF Research Database (Denmark)

    Josefsen, Knud; Lee, Ying Chiu

    2011-01-01

    Real time PCR is the analytic tool of choice for quantification of gene expression, while RNAi is concerned with downregulation of gene expression. Together, they constitute a powerful approach in any loss of function studies of selective genes. We illustrate here the use of real time PCR to verify...

  5. Bioinformatic tools for PCR Primer design

    African Journals Online (AJOL)

    ES

    reaction (PCR), oligo hybridization and DNA sequencing. Proper primer design is actually one of the most important factors/steps in successful DNA sequencing. Various bioinformatics programs are available for selection of primer pairs from a template sequence. The plethora programs for PCR primer design reflects the.

  6. Comparative analysis of conventional PCR and real-time PCR to diagnose shrimp WSD

    Directory of Open Access Journals (Sweden)

    C.A.G. Leal

    2013-09-01

    Full Text Available The aims of this study were to standard and optimize a qPCR protocol with FAM-BHQ1 probe, and to compare its sensitivity against TaqMan qPCR and PCR methods to diagnose shrimp WSD. The FAM-BHQ1 qPCR presented higher clinical sensitivity and showed to be a robust alternative to detect WSSV in clinical samples.

  7. Digital electrostatic acoustic transducer array

    KAUST Repository

    Carreno, Armando Arpys Arevalo

    2016-12-19

    In this paper we present the fabrication and characterization of an array of electrostatic acoustic transducers. The array is micromachined on a silicon wafer using standard micro-machining techniques. Each array contains 2n electrostatic transducer membranes, where “n” is the bit number. Every element of the array has a hexagonal membrane shape structure, which is separated from the substrate by 3µm air gap. The membrane is made out 5µm thick polyimide layer that has a bottom gold electrode on the substrate and a gold top electrode on top of the membrane (250nm). The wafer layout design was diced in nine chips with different array configurations, with variation of the membrane dimensions. The device was tested with 90 V giving and sound output level as high as 35dB, while actuating all the elements at the same time.

  8. Chunking of Large Multidimensional Arrays

    Energy Technology Data Exchange (ETDEWEB)

    Rotem, Doron; Otoo, Ekow J.; Seshadri, Sridhar

    2007-02-28

    Data intensive scientific computations as well on-lineanalytical processing applications as are done on very large datasetsthat are modeled as k-dimensional arrays. The storage organization ofsuch arrays on disks is done by partitioning the large global array intofixed size hyper-rectangular sub-arrays called chunks or tiles that formthe units of data transfer between disk and memory. Typical queriesinvolve the retrieval of sub-arrays in a manner that accesses all chunksthat overlap the query results. An important metric of the storageefficiency is the expected number of chunks retrieved over all suchqueries. The question that immediately arises is "what shapes of arraychunks give the minimum expected number of chunks over a query workload?"In this paper we develop two probabilistic mathematical models of theproblem and provide exact solutions using steepest descent and geometricprogramming methods. Experimental results, using synthetic workloads onreal life data sets, show that our chunking is much more efficient thanthe existing approximate solutions.

  9. Polymerase chain reaction methods (PCR in agrobiotechnology

    Directory of Open Access Journals (Sweden)

    Taški-Ajduković Ksenija

    2006-01-01

    Full Text Available The agricultural biotechnology applies polymerase chain reaction (PCR technology at numerous steps throughout product development. The major uses of PCR technology during product development include gene discovery and cloning, vector construction, transformant identification, screening and characterization as well as seed quality control. Commodity and food companies as well as testing laboratories rely on PCR technology to verify the presence or absence of genetically modification (GM in a product or to quantify the amount of GM material present in the product. This article describes the fundamental elements of PCR analysis and its application to the testing of grains and highlights some of areas to which attention must be paid in order to produce reliable test results. The article also discuses issues related to the analysis of different matrixes and the effect they may have on the accuracy of the PCR analytical results.

  10. High speed single nucleotide polymorphism typing of a hereditary haemochromatosis mutation with capillary array electrophoresis microplates.

    Science.gov (United States)

    Medintz, I; Wong, W W; Sensabaugh, G; Mathies, R A

    2000-07-01

    A single nucleotide polymorphism (SNP) typing assay is developed and evaluated on a microfabricated capillary array electrophoresis system. Using fluorescently labeled allele-specific primers, the S65C (193A-->T) substitution associated with hereditary haemochromatosis in the HFE gene is genotyped. The covalently labeled polymerase chain reaction (PCR) products are separated on a microfabricated radial capillary array electrophoresis microplate using nondenaturing gel media in under two minutes. Detection is accomplished with a laser-excited rotary confocal scanner. The Rox-labeled A-allele specific amplicon (211 bp) is differentiated from the R110-labeled T-allele specific amplicon (201 bp) by both size and color. This study demonstrates the feasibility of using allele-specific PCR with covalently labeled primers for high speed fluorescent SNP typing on microfabricated radial capillary array electrophoresis microplates.

  11. Planning a Global Array of Broadband Seismic Arrays

    Science.gov (United States)

    Koper, Keith D.; Ammon, Charles J.

    2013-08-01

    A diverse group of more than 70 seismologists met for 2 days in Raleigh, N.C., to report on recent innovations in seismic array methods and to discuss the future of seismic arrays in global seismology. The workshop was sponsored by the Incorporated Research Institutions for Seismology (IRIS), with U.S. National Science Foundation funding. Participants included representatives of existing array research groups in Australia, Canada, Germany, Japan, Norway, and the United States, with individuals from academia, government, and industry. The workshop was organized by the authors of this meeting report, Pablo Ampeuro (California Institute of Technology), and Colleen Dalton (Boston University), along with IRIS staff support.

  12. Dependently typed array programs don't go wrong

    NARCIS (Netherlands)

    Trojahner, K.; Grelck, C.

    2008-01-01

    The array programming paradigm adopts multidimensional arrays as the fundamental data structures of computation. Array operations process entire arrays instead of just single elements. This makes array programs highly expressive and introduces data parallelism in a natural way. Array programming

  13. Dependently typed array programs don’t go wrong

    NARCIS (Netherlands)

    Trojahner, K.; Grelck, C.

    2009-01-01

    The array programming paradigm adopts multidimensional arrays as the fundamental data structures of computation. Array operations process entire arrays instead of just single elements. This makes array programs highly expressive and introduces data parallelism in a natural way. Array programming

  14. Evaluation of PCR and nested PCR for laboratory diagnosis of hepatitis C virus infection.

    Science.gov (United States)

    Aslanzadeh, J; Padilla, B B; Shanley, J D

    1996-06-01

    The detection of hepatitis C virus (HCV) RNA by nested polymerase chain reaction (PCR) is believed to be the most reliable method to diagnose HCV infections. A pitfall of nested PCR is that it is prone to contamination. Single step reverse transcription-PCR (RT-PCR) was performed, prospectively, on 80 sera from 59 patients with a set of primers that amplified a 273 bp sequence unique to the 5' noncoding (NC) region of the HCV genome. Nested PCR, was performed on all PCR negative specimens with a set of primers that amplified a 255 bp internal to the original primers. Single step RT-PCR was positive on 45 sera from 35 patients following gel electrophoresis and on two additional sera from two patients following Southern blot hybridization. Nested PCR was positive on two more sera following gel electrophoresis of the nested PCR products. These two patients were seropositive and subsequent serum from one patient was positive by single step PCR. Three additional sera were positive following Southern blot analysis of the nested PCR products. Two patients were seropositive and had elevated serum alanine aminotransferase (ALT) levels. The third patient was seronegative with normal ALT level and was considered a false positive. The remaining seronegative control specimens were PCR negative by both methods. The majority of PCR positive patients (82%) had elevated ALT levels, while the majority of PCR negative seropositive patients had normal ALT levels. We conclude that single step PCR is a sensitive test for the laboratory diagnoses of the majority of the HCV infections.

  15. ESPRIT And Uniform Linear Arrays

    Science.gov (United States)

    Roy, R. H.; Goldburg, M.; Ottersten, B. E.; Swindlehurst, A. L.; Viberg, M.; Kailath, T.

    1989-11-01

    Abstract ¬â€?ESPRIT is a recently developed and patented technique for high-resolution estimation of signal parameters. It exploits an invariance structure designed into the sensor array to achieve a reduction in computational requirements of many orders of magnitude over previous techniques such as MUSIC, Burg's MEM, and Capon's ML, and in addition achieves performance improvement as measured by parameter estimate error variance. It is also manifestly more robust with respect to sensor errors (e.g. gain, phase, and location errors) than other methods as well. Whereas ESPRIT only requires that the sensor array possess a single invariance best visualized by considering two identical but other-wise arbitrary arrays of sensors displaced (but not rotated) with respect to each other, many arrays currently in use in various applications are uniform linear arrays of identical sensor elements. Phased array radars are commonplace in high-resolution direction finding systems, and uniform tapped delay lines (i.e., constant rate A/D converters) are the rule rather than the exception in digital signal processing systems. Such arrays possess many invariances, and are amenable to other types of analysis, which is one of the main reasons such structures are so prevalent. Recent developments in high-resolution algorithms of the signal/noise subspace genre including total least squares (TLS) ESPRIT applied to uniform linear arrays are summarized. ESPRIT is also shown to be a generalization of the root-MUSIC algorithm (applicable only to the case of uniform linear arrays of omni-directional sensors and unimodular cisoids). Comparisons with various estimator bounds, including CramerRao bounds, are presented.

  16. Material Biocompatibility for PCR Microfluidic Chips

    KAUST Repository

    Kodzius, Rimantas

    2010-04-23

    As part of the current miniaturization trend, biological reactions and processes are being adapted to microfluidics devices. PCR is the primary method employed in DNA amplification, its miniaturization is central to efforts to develop portable devices for diagnostics and testing purposes. A problem is the PCR-inhibitory effect due to interaction between PCR reagents and the surrounding environment, which effect is increased in high-surface-are-to-volume ration microfluidics. In this study, we evaluated the biocompatibility of various common materials employed in the fabrication of microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most of the cases, addition of bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, whereas they did show noticeable interaction with the DNA polymerase. Our test, instead of using microfluidic devices, can be easily conducted in common PCR tubes using a standard bench thermocycler. Our data supports an overview of the means by which the materials most bio-friendly to microfluidics can be selected.

  17. Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk.

    Science.gov (United States)

    Wang, Meng; Yang, Junjie; Gai, Zhongtao; Huo, Shengnan; Zhu, Jianhua; Li, Jun; Wang, Ranran; Xing, Sheng; Shi, Guosheng; Shi, Feng; Zhang, Lei

    2018-02-02

    As a kind of zero-tolerance foodborne pathogens, Salmonella typhimurium poses a great threat to quality of food products and public health. Hence, rapid and efficient approaches to identify Salmonella typhimurium are urgently needed. Combined with PCR and fluorescence technique, real-time PCR (qPCR) and digital PCR (ddPCR) are regarded as suitable tools for detecting foodborne pathogens. To compare the effect between qPCR and ddPCR in detecting Salmonella typhimurium, a series of nucleic acid, pure strain culture and spiking milk samples were applied and the resistance to inhibitors referred in this article as well. Compared with qPCR, ddPCR exhibited more sensitive (10 -4 ng/μl or 10 2 cfu/ml) and less pre-culturing time (saving 2h). Moreover, ddPCR had stronger resistance to inhibitors than qPCR, yet absolute quantification hardly performed when target's concentration over 1ng/μl or 10 6 cfu/ml. This study provides an alternative strategy in detecting foodborne Salmonella typhimurium. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Fundamentals of ultrasonic phased arrays

    CERN Document Server

    Schmerr, Lester W

    2014-01-01

    This book describes in detail the physical and mathematical foundations of ultrasonic phased array measurements.?The book uses linear systems theory to develop a comprehensive model of the signals and images that can be formed with phased arrays. Engineers working in the field of ultrasonic nondestructive evaluation (NDE) will find in this approach a wealth of information on how to design, optimize and interpret ultrasonic inspections with phased arrays. The fundamentals and models described in the book will also be of significant interest to other fields, including the medical ultrasound and

  19. Antenna arrays a computational approach

    CERN Document Server

    Haupt, Randy L

    2010-01-01

    This book covers a wide range of antenna array topics that are becoming increasingly important in wireless applications, particularly in design and computer modeling. Signal processing and numerical modeling algorithms are explored, and MATLAB computer codes are provided for many of the design examples. Pictures of antenna arrays and components provided by industry and government sources are presented with explanations of how they work. Antenna Arrays is a valuable reference for practicing engineers and scientists in wireless communications, radar, and remote sensing, and an excellent textbook for advanced antenna courses.

  20. Interferometric optical vortex array generator.

    Science.gov (United States)

    Vyas, Sunil; Senthilkumaran, P

    2007-05-20

    Two new interferometric configurations for optical vortex array generation are presented. These interferometers are different from the conventional interferometers in that they are capable of producing a large number of isolated zeros of intensity, and all of them contain optical vortices. Simulation and theory for optical vortex array generation using three-plane-wave interference is presented. The vortex dipole array produced this way is noninteracting, as there are no attraction or repulsion forces between them, leading to annihilation or creation of vortex pairs.

  1. On the Mesh Array for Matrix Multiplication

    OpenAIRE

    Kak, Subhash

    2010-01-01

    This article presents new properties of the mesh array for matrix multiplication. In contrast to the standard array that requires 3n-2 steps to complete its computation, the mesh array requires only 2n-1 steps. Symmetries of the mesh array computed values are presented which enhance the efficiency of the array for specific applications. In multiplying symmetric matrices, the results are obtained in 3n/2+1 steps. The mesh array is examined for its application as a scrambling system.

  2. Next Generation Microshutter Arrays Project

    Data.gov (United States)

    National Aeronautics and Space Administration — We propose to develop the next generation MicroShutter Array (MSA) as a multi-object field selector for missions anticipated in the next two decades. For many...

  3. Fundamentals of spherical array processing

    CERN Document Server

    Rafaely, Boaz

    2015-01-01

    This book provides a comprehensive introduction to the theory and practice of spherical microphone arrays. It is written for graduate students, researchers and engineers who work with spherical microphone arrays in a wide range of applications.   The first two chapters provide the reader with the necessary mathematical and physical background, including an introduction to the spherical Fourier transform and the formulation of plane-wave sound fields in the spherical harmonic domain. The third chapter covers the theory of spatial sampling, employed when selecting the positions of microphones to sample sound pressure functions in space. Subsequent chapters present various spherical array configurations, including the popular rigid-sphere-based configuration. Beamforming (spatial filtering) in the spherical harmonics domain, including axis-symmetric beamforming, and the performance measures of directivity index and white noise gain are introduced, and a range of optimal beamformers for spherical arrays, includi...

  4. Thermopile Area Array Readout Project

    Data.gov (United States)

    National Aeronautics and Space Administration — NASA/JPL thermopile detector linear arrays, wire bonded to Black Forest Engineering (BFE) CMOS readout integrated circuits (ROICs), have been utilized in NASA...

  5. [Identification of 22q11.2 microdeletion among patients with congenital heart diseases using droplet digital PCR].

    Science.gov (United States)

    Zhou, Xiangcheng; Zhang, Cuicui; Li, Mi; Ma, Jian; Chen, Qiuping; Sun, Shanquan; Zhang, Liang

    2018-02-10

    OBJECTIVE To develop a new method for detecting 22q11.2 deletion syndrome (22q11.2 DS) in clinical settings. METHODS Specific primers and fluorescence probes were designed to target the TBX1 gene within the 22q11.2 deletion region and a reference gene RPP30. Multiplexed droplet digital PCR (ddPCR) was run to detect the 22q11.2 microdeletion by calculating the ratio of positive droplet number of TBX1/RPP30. RESULTS Three cases of 22q11.2 microdeletion previously confirmed by array comparative genome hybridization were successfully identified. Subsequently, the ddPCR detected two further cases of 22q11.2 microdeletion among 14 children with congenital heart diseases. CONCLUSION The ddPCR technique has provided a rapid and cost-effective method for detecting 22q11.2 microdeletion in clinical settings.

  6. The Applicability of Incoherent Array Processing to IMS Seismic Arrays

    Science.gov (United States)

    Gibbons, Steven J.

    2014-03-01

    The seismic arrays of the International Monitoring System (IMS) for the Comprehensive Nuclear-Test-Ban Treaty (CTBT) are highly diverse in size and configuration, with apertures ranging from under 1 km to over 60 km. Large and medium aperture arrays with large inter-site spacings complicate the detection and estimation of high-frequency phases lacking coherence between sensors. Pipeline detection algorithms often miss such phases, since they only consider frequencies low enough to allow coherent array processing, and phases that are detected are often attributed qualitatively incorrect backazimuth and slowness estimates. This can result in missed events, due to either a lack of contributing phases or by corruption of event hypotheses by spurious detections. It has been demonstrated previously that continuous spectral estimation can both detect and estimate phases on the largest aperture arrays, with arrivals identified as local maxima on beams of transformed spectrograms. The estimation procedure in effect measures group velocity rather than phase velocity, as is the case for classical f-k analysis, and the ability to estimate slowness vectors requires sufficiently large inter-sensor distances to resolve time-delays between pulses with a period of the order 4-5 s. Spectrogram beampacking works well on five IMS arrays with apertures over 20 km (NOA, AKASG, YKA, WRA, and KURK) without additional post-processing. Seven arrays with 10-20 km aperture (MJAR, ESDC, ILAR, KSRS, CMAR, ASAR, and EKA) can provide robust parameter estimates subject to a smoothing of the resulting slowness grids, most effectively achieved by convolving the measured slowness grids with the array response function for a 4 or 5 s period signal. Even for medium aperture arrays which can provide high-quality coherent slowness estimates, a complementary spectrogram beampacking procedure could act as a quality control by providing non-aliased estimates when the coherent slowness grids display

  7. The potential advantages of digital PCR for clinical virology diagnostics.

    Science.gov (United States)

    Hall Sedlak, Ruth; Jerome, Keith R

    2014-05-01

    Digital PCR (dPCR), a new nucleic acid amplification technology, offers several potential advantages over real-time or quantitative PCR (qPCR), the current workhorse of clinical molecular virology diagnostics. Several studies have demonstrated dPCR assays for human cytomegalovirus or HIV, which give more precise and reproducible results than qPCR assays without sacrificing sensitivity. Here we review the literature comparing dPCR and qPCR performance in viral molecular diagnostic assays and offer perspective on the future of dPCR in clinical virology diagnostics.

  8. Calibrated user-friendly reverse transcriptase-PCR assay

    DEFF Research Database (Denmark)

    Bor, M V; Sørensen, B S; Rammer, P

    1998-01-01

    We report a competitive reverse transcriptase-PCR (RT-PCR) assay and a calibrated user-friendly RT-PCR assay (CURT-PCR) for epidermal growth factor receptor (EGFR) mRNA. A calibrator was prepared from isolated rat liver RNA, and the amount of EGFR mRNA was determined by competitive RT-PCR. In CURT...

  9. Flexible eddy current coil arrays

    International Nuclear Information System (INIS)

    Krampfner, Y.; Johnson, D.P.

    1987-01-01

    A novel approach was devised to overcome certain limitations of conventional eddy current testing. The typical single-element hand-wound probe was replaced with a two dimensional array of spirally wound probe elements deposited on a thin, flexible polyimide substrate. This provides full and reliable coverage of the test area and eliminates the need for scanning. The flexible substrate construction of the array allows the probes to conform to irregular part geometries, such as turbine blades and tubing, thereby eliminating the need for specialized probes for each geometry. Additionally, the batch manufacturing process of the array can yield highly uniform and reproducible coil geometries. The array is driven by a portable computer-based eddy current instrument, smartEDDY/sup TM/, capable of two-frequency operation, and offers a great deal of versatility and flexibility due to its software-based architecture. The array is coupled to the instrument via an 80-switch multiplexer that can be configured to address up to 1600 probes. The individual array elements may be addressed in any desired sequence, as defined by the software

  10. Calibration of a microprobe array

    International Nuclear Information System (INIS)

    Schrader, Christian; Tutsch, Rainer

    2012-01-01

    Conventional coordinate measurement machines are not well adapted to the specific needs for the measurement of mechanical microstructures that are made in a highly parallel production process. In particular, the increase of the measurement speed is addressed by using an array of microprobes to measure a number of objects in parallel. It consists of multiple microprobes that are etched into the same silicon substrate. The styli are glued onto a boss structure in the middle of a silicon membrane. To facilitate the alignment of an array and the underlying wafer, the array is mounted on three stacked rotational stages. Due to the production tolerances, the positions of the touching balls of the probes relative to their pivot have to be calibrated. The probe sensitivity is another field of calibration. This paper describes an efficient calibration procedure of the probe array which is usable for arrays with a large number of probes and different array layouts. The validation method of this procedure is explained and calibration results are discussed (paper)

  11. Integrated residential photovoltaic array development

    Science.gov (United States)

    Royal, G. C., III

    1981-04-01

    Sixteen conceptual designs of residential photovoltaic arrays are described. Each design concept was evaluated by an industry advisory panel using a comprehensive set of technical, economic and institutional criteria. Key electrical and mechanical concerns that effect further array subsystem development are also discussed. Three integrated array design concepts were selected by the advisory panel for further optimization and development. From these concepts a single one will be selected for detailed analysis and prototype fabrication. The three concepts selected are: (1) An array of frameless panels/modules sealed in a T shaped zipper locking neoprene gasket grid pressure fitted into an extruded aluminum channel grid fastened across the rafters. (2) An array of frameless modules pressure fitted in a series of zipper locking EPDM rubber extrusions adhesively bonded to the roof. Series string voltage is developed using a set of integral tongue connectors and positioning blocks. (3) An array of frameless modules sealed by a silicone adhesive in a prefabricated grid of rigid tape and sheet metal attached to the roof.

  12. CCD and IR array controllers

    Science.gov (United States)

    Leach, Robert W.; Low, Frank J.

    2000-08-01

    A family of controllers has bene developed that is powerful and flexible enough to operate a wide range of CCD and IR focal plane arrays in a variety of ground-based applications. These include fast readout of small CCD and IR arrays for adaptive optics applications, slow readout of large CCD and IR mosaics, and single CCD and IR array operation at low background/low noise regimes as well as high background/high speed regimes. The CCD and IR controllers have a common digital core based on user- programmable digital signal processors that are used to generate the array clocking and signal processing signals customized for each application. A fiber optic link passes image data and commands to VME or PCI interface boards resident in a host computer to the controller. CCD signal processing is done with a dual slope integrator operating at speeds of up to one Megapixel per second per channel. Signal processing of IR arrays is done either with a dual channel video processor or a four channel video processor that has built-in image memory and a coadder to 32-bit precision for operating high background arrays. Recent developments underway include the implementation of a fast fiber optic data link operating at a speed of 12.5 Megapixels per second for fast image transfer from the controller to the host computer, and supporting image acquisition software and device drivers for the PCI interface board for the Sun Solaris, Linux and Windows 2000 operating systems.

  13. A naked-eye colorimetric "PCR developer"

    Science.gov (United States)

    Valentini, Paola; Pompa, Pier Paolo

    2016-04-01

    Despite several advances in molecular biology and diagnostics, Polymerase Chain Reaction (PCR) is currently the gold standard for nucleic acids amplification and detection, due to its versatility, low-cost and universality, with estimated market of several billion dollars/year. Nevertheless, PCR still relies on the laborious, time-consuming, and multi-step gel electrophoresis-based detection, which includes gel casting, electrophoretic run, gel staining, and gel visualization. In this work, we propose a "PCR developer", namely a universal one-step, one-tube method, based on controlled aggregation of gold nanoparticles (AuNPs), to detect PCR products by naked eye in few minutes, with no need for any instrumentation. We demonstrated the specificity and sensitivity of the PCR developer on different model targets, suitable for a qualitative detection in real-world diagnostics (i.e., gene rearrangements, genetically modified organisms, and pathogens). The PCR developer proved to be highly specific and ultra-sensitive, discriminating down to few copies of HIV viral DNA, diluted in an excess of interfering human genomic DNA, which is a clinically relevant viral load. Hence, it could be a valuable tool for both academic research and clinical applications.

  14. Toward single enzyme analysis in a droplet-based micro and nanofluidic system

    NARCIS (Netherlands)

    Arayanarakool, Rerngchai

    2012-01-01

    In this thesis, we have demonstrated the application of micro- and nanofluidic devices to generate an array of aqueous droplets in oil phase for single-enzyme encapsulation and activity measurement. We chose droplet-based microfluidics for this purpose of monitoring single-enzyme reactions since the

  15. MassCode Liquid Arrays as a Tool for Multiplexed High-Throughput Genetic Profiling

    Science.gov (United States)

    Richmond, Gregory S.; Khine, Htet; Zhou, Tina T.; Ryan, Daniel E.; Brand, Tony; McBride, Mary T.; Killeen, Kevin

    2011-01-01

    Multiplexed detection assays that analyze a modest number of nucleic acid targets over large sample sets are emerging as the preferred testing approach in such applications as routine pathogen typing, outbreak monitoring, and diagnostics. However, very few DNA testing platforms have proven to offer a solution for mid-plexed analysis that is high-throughput, sensitive, and with a low cost per test. In this work, an enhanced genotyping method based on MassCode technology was devised and integrated as part of a high-throughput mid-plexing analytical system that facilitates robust qualitative differential detection of DNA targets. Samples are first analyzed using MassCode PCR (MC-PCR) performed with an array of primer sets encoded with unique mass tags. Lambda exonuclease and an array of MassCode probes are then contacted with MC-PCR products for further interrogation and target sequences are specifically identified. Primer and probe hybridizations occur in homogeneous solution, a clear advantage over micro- or nanoparticle suspension arrays. The two cognate tags coupled to resultant MassCode hybrids are detected in an automated process using a benchtop single quadrupole mass spectrometer. The prospective value of using MassCode probe arrays for multiplexed bioanalysis was demonstrated after developing a 14plex proof of concept assay designed to subtype a select panel of Salmonella enterica serogroups and serovars. This MassCode system is very flexible and test panels can be customized to include more, less, or different markers. PMID:21544191

  16. MassCode liquid arrays as a tool for multiplexed high-throughput genetic profiling.

    Directory of Open Access Journals (Sweden)

    Gregory S Richmond

    Full Text Available Multiplexed detection assays that analyze a modest number of nucleic acid targets over large sample sets are emerging as the preferred testing approach in such applications as routine pathogen typing, outbreak monitoring, and diagnostics. However, very few DNA testing platforms have proven to offer a solution for mid-plexed analysis that is high-throughput, sensitive, and with a low cost per test. In this work, an enhanced genotyping method based on MassCode technology was devised and integrated as part of a high-throughput mid-plexing analytical system that facilitates robust qualitative differential detection of DNA targets. Samples are first analyzed using MassCode PCR (MC-PCR performed with an array of primer sets encoded with unique mass tags. Lambda exonuclease and an array of MassCode probes are then contacted with MC-PCR products for further interrogation and target sequences are specifically identified. Primer and probe hybridizations occur in homogeneous solution, a clear advantage over micro- or nanoparticle suspension arrays. The two cognate tags coupled to resultant MassCode hybrids are detected in an automated process using a benchtop single quadrupole mass spectrometer. The prospective value of using MassCode probe arrays for multiplexed bioanalysis was demonstrated after developing a 14plex proof of concept assay designed to subtype a select panel of Salmonella enterica serogroups and serovars. This MassCode system is very flexible and test panels can be customized to include more, less, or different markers.

  17. SASqPCR: robust and rapid analysis of RT-qPCR data in SAS.

    Directory of Open Access Journals (Sweden)

    Daijun Ling

    Full Text Available Reverse transcription quantitative real-time PCR (RT-qPCR is a key method for measurement of relative gene expression. Analysis of RT-qPCR data requires many iterative computations for data normalization and analytical optimization. Currently no computer program for RT-qPCR data analysis is suitable for analytical optimization and user-controllable customization based on data quality, experimental design as well as specific research aims. Here I introduce an all-in-one computer program, SASqPCR, for robust and rapid analysis of RT-qPCR data in SAS. This program has multiple macros for assessment of PCR efficiencies, validation of reference genes, optimization of data normalizers, normalization of confounding variations across samples, and statistical comparison of target gene expression in parallel samples. Users can simply change the macro variables to test various analytical strategies, optimize results and customize the analytical processes. In addition, it is highly automatic and functionally extendable. Thus users are the actual decision-makers controlling RT-qPCR data analyses. SASqPCR and its tutorial are freely available at http://code.google.com/p/sasqpcr/downloads/list.

  18. Signal and noise in bridging PCR

    Directory of Open Access Journals (Sweden)

    Thaler David S

    2002-07-01

    Full Text Available Abstract Background In a variant of the standard PCR reaction termed bridging, or jumping, PCR the primer-bound sequences are originally on separate template molecules. Bridging can occur if, and only if, the templates contain a region of sequence similarity. A 3' end of synthesis in one round of synthesis that terminates in this region of similarity can prime on the other. In principle, Bridging PCR (BPCR can detect a subpopulation of one template that terminates synthesis in the region of sequence shared by the other template. This study considers the sensitivity and noise of BPCR as a quantitative assay for backbone interruptions. Bridging synthesis is also important to some methods for computing with DNA. Results In this study, BPCR was tested over a 328 base pair segment of the E. coli lac operon and a signal to noise ratio (S/N of approximately 10 was obtained under normal PCR conditions with Taq polymerase. With special precautions in the case of Taq or by using the Stoffel fragment the S/N was improved to 100, i.e. 1 part of cut input DNA yielded the same output as 100 parts of intact input DNA. Conclusions In the E. coli lac operator region studied here, depending on details of protocol, between 3 and 30% per kilobase of final PCR product resulted from bridging. Other systems are expected to differ in the proportion of product that is bridged consequent to PCR protocol and the sequence analyzed. In many cases physical bridging during PCR will have no informational consequence because the bridged templates are of identical sequence, but in a number of special cases bridging creates, or, destroys, information.

  19. Coded aperture imaging with uniformly redundant arrays

    International Nuclear Information System (INIS)

    Fenimore, E.E.; Cannon, T.M.

    1980-01-01

    A system is described which uses uniformly redundant arrays to image non-focusable radiation. The array is used in conjunction with a balanced correlation technique to provide a system with no artifacts so that virtually limitless signal-to-noise ratio is obtained with high transmission characteristics. The array is mosaicked to reduce required detector size over conventional array detectors. 15 claims

  20. Retrieval of Mir Solar Array

    Science.gov (United States)

    Rutledge, Sharon K.; deGroh, Kim K.

    1999-01-01

    A Russian solar array panel removed in November 1997 from the non-articulating photovoltaic array on the Mir core module was returned to Earth on STS-89 in January 1998. The panel had been exposed to low Earth orbit (LEO) for 10 years prior to retrieval. The retrieval provided a unique opportunity to study the effects of the LEO environment on a functional solar array. To take advantage of this opportunity, a team composed of members from RSC-Energia (Russia), the Boeing Company, and the following NASA Centers--Johnson Space Center, Kennedy Space Center, Langley Research Center, Marshall Space Flight Center, and Lewis Research Center--was put together to analyze the array. After post-retrieval inspections at the Spacehab Facility at Kennedy in Florida, the array was shipped to Lewis in Cleveland for electrical performance tests, closeup photodocumentation, and removal of selected solar cells and blanket material. With approval from RSC-Energia, five cell pairs and their accompanying blanket and mesh material, and samples of painted handrail materials were selected for removal on the basis of their ability to provide degradation information. Sites were selected that provided different sizes and shapes of micrometeoroid impacts and different levels of surface contamination. These materials were then distributed among the team for round robin testing.

  1. The principle and application of new PCR Technologies

    Science.gov (United States)

    Yu, Miao; Cao, Yue; Ji, Yubin

    2017-12-01

    Polymerase chain reaction (PCR) is essentially a selective DNA amplification technique commonlyapplied for genetic testing and molecular diagnosis because of its high specificity and sensitivity.PCR technologies as the key of molecular biology, has realized that the qualitative detection of absolute quantitative has been changed. It has produced a variety of new PCR technologies, such as extreme PCR, photonic PCR, o-amplification at lower denaturation temperature PCR, nanoparticle PCR and so on. In this paper, the principle and application of PCR technologies are reviewed, and its development is prospected too.

  2. PCR+ In Diesel Fuels and Emissions Research

    Energy Technology Data Exchange (ETDEWEB)

    McAdams, H.T.

    2002-04-15

    In past work for the U.S. Department of Energy (DOE) and Oak Ridge National Laboratory (ORNL), PCR+ was developed as an alternative methodology for building statistical models. PCR+ is an extension of Principal Components Regression (PCR), in which the eigenvectors resulting from Principal Components Analysis (PCA) are used as predictor variables in regression analysis. The work was motivated by the observation that most heavy-duty diesel (HDD) engine research was conducted with test fuels that had been ''concocted'' in the laboratory to vary selected fuel properties in isolation from each other. This approach departs markedly from the real world, where the reformulation of diesel fuels for almost any purpose leads to changes in a number of interrelated properties. In this work, we present new information regarding the problems encountered in the conventional approach to model-building and how the PCR+ method can be used to improve research on the relationship between fuel characteristics and engine emissions. We also discuss how PCR+ can be applied to a variety of other research problems related to diesel fuels.

  3. X-ray detector array

    International Nuclear Information System (INIS)

    Houston, J.M.

    1980-01-01

    The object of the invention (an ionization chamber X-ray detector array for use with high speed computerised tomographic imaging apparatus) is to reduce the time required to produce a tomographic image. The detector array described determines the distribution of X-ray intensities in one or more flat, coplanar X-ray beams. It comprises three flat anode sheets parallel to the X-ray beam, a plurality of rod-like cathodes between the anodes, a detector gas between the electrodes and a means for applying a potential between the electrodes. Each of the X-ray sources is collimated to give a narrow, planar section of X-ray photons. Sets of X-ray sources in the array are pulsed simultaneously to obtain X-ray transmission data for tomographic image reconstruction. (U.K.)

  4. Innovations in IR projector arrays

    Science.gov (United States)

    Cole, Barry E.; Higashi, B.; Ridley, Jeff A.; Holmen, J.; Newstrom, K.; Zins, C.; Nguyen, K.; Weeres, Steven R.; Johnson, Burgess R.; Stockbridge, Robert G.; Murrer, Robert Lee; Olson, Eric M.; Bergin, Thomas P.; Kircher, James R.; Flynn, David S.

    2000-07-01

    In the past year, Honeywell has developed a 512 X 512 snapshot scene projector containing pixels with very high radiance efficiency. The array can operate in both snapshot and raster mode. The array pixels have near black body characteristics, high radiance outputs, broad band performance, and high speed. IR measurements and performance of these pixels will be described. In addition, a vacuum probe station that makes it possible to select the best die for packaging and delivery based on wafer level radiance screening, has been developed and is in operation. This system, as well as other improvements, will be described. Finally, a review of the status of the present projectors and plans for future arrays is included.

  5. Thin, Flexible IMM Solar Array

    Science.gov (United States)

    Walmsley, Nicholas

    2015-01-01

    NASA needs solar arrays that are thin, flexible, and highly efficient; package compactly for launch; and deploy into large, structurally stable high-power generators. Inverted metamorphic multijunction (IMM) solar cells can enable these arrays, but integration of this thin crystalline cell technology presents certain challenges. The Thin Hybrid Interconnected Solar Array (THINS) technology allows robust and reliable integration of IMM cells into a flexible blanket comprising standardized modules engineered for easy production. The modules support the IMM cell by using multifunctional materials for structural stability, shielding, coefficient of thermal expansion (CTE) stress relief, and integrated thermal and electrical functions. The design approach includes total encapsulation, which benefits high voltage as well as electrostatic performance.

  6. A bead-based suspension array for the multiplexed detection of begomoviruses and their whitefly vectors.

    Science.gov (United States)

    van Brunschot, S L; Bergervoet, J H W; Pagendam, D E; de Weerdt, M; Geering, A D W; Drenth, A; van der Vlugt, R A A

    2014-03-01

    Bead-based suspension array systems enable simultaneous fluorescence-based identification of multiple nucleic acid targets in a single reaction. This study describes the development of a novel approach to plant virus and vector diagnostics, a multiplexed 7-plex array that comprises a hierarchical set of assays for the simultaneous detection of begomoviruses and Bemisia tabaci, from both plant and whitefly samples. The multiplexed array incorporates genus, species and strain-specific assays, offering a unique approach for identifying both known and unknown viruses and B. tabaci species. When tested against a large panel of sequence-characterized begomovirus and whitefly samples, the array was shown to be 100% specific to the homologous target. Additionally, the multiplexed array was highly sensitive, efficiently and concurrently determining both virus and whitefly identity from single viruliferous whitefly samples. The detection limit for one assay within the multiplexed array that specifically detects Tomato yellow leaf curl virus-Israel (TYLCV-IL) was quantified as 200fg of TYLCV-IL DNA, directly equivalent to that of TYLCV-specific qPCR. Highly reproducible results were obtained over multiple tests. The flexible multiplexed array described in this study has great potential for use in plant quarantine, biosecurity and disease management programs worldwide. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. From the 'PCR' function to the 'PCR' profession; de la fonction 'PCR' au metier 'PCR'

    Energy Technology Data Exchange (ETDEWEB)

    Perrin, L. [CERAP, 91 - Gif sur Yvette (France)

    2008-07-01

    After having recalled the legal context concerning the appointment and training of a radiation protection expert (PCR for 'personne competente en radioprotection'), the author outlines that the PCR's role has notably evolved: his function is now of primary importance in the company and his activity does not correspond to the legal framework any longer. Moreover, with the application of a European directive, some small establishments possessing ionizing radiation sources are disadvantaged, and the PCR is now facing an increasing number of missions and tasks. The author gives a list of them and assesses a needed time of 146 days per year: this means PCRs cannot have an other activity within their company

  8. Broadband acoustic phased array with subwavelength active tube array

    Science.gov (United States)

    Li, Xiao-Yan; Yang, Zhang-Zhao; Zhu, Yi-Fan; Zou, Xin-Ye; Cheng, Jian-Chun

    2018-02-01

    Acoustic metasurfaces provide a way to manipulate wavefronts at anomalous reflection or refraction angles through subwavelength structures. Here, based on the generalized Snell's refraction law for acoustic metasurfaces and the classical acoustic phased array (PA) theory, a broadband acoustic PA with a subwavelength active tube array has been proposed to form a special acoustic beam and to determine the directivity characteristics of the acoustic source. Theoretical analysis shows that the dispersionless wavefront manipulation can be realized by the gradient model of the active tube array, and a wide working frequency band can be obtained in practical applications from the simulated and experimental results. The numerical results of forming a special acoustic beam and establishing an acoustic focus model with an arbitrary focal position are consistent with the theoretical predictions. The experimental results agree well with the simulated results in the model of forming the acoustic beam of 45 ° . By combining acoustic metamaterials and conventional acoustic PA, the model of the active tube array paves a way to design a composite acoustic PA with high radiation efficiency and system robustness without the need for any complex circuit control system. This design concept is expected to be used in the design of ultrasonic therapy devices and high-efficiency transducers.

  9. Phased arrays: inline flow line hub inspection using phased arrays

    NARCIS (Netherlands)

    Bloom, J.G.P.; Chougrani, K.; Rundberg, H.; Oldenziel, G.; Deleye, X.; Martina, Q.

    2011-01-01

    The feasibility of the inspection of flow line hubs using the phased array technique was investigated to determine the surface area of the seal area degraded by corrosion. A clean model of the hub was simulated to gain insight into the geometrical echoes and to determine the area covered by the

  10. Terahertz superconducting plasmonic hole array.

    Science.gov (United States)

    Tian, Zhen; Singh, Ranjan; Han, Jiaguang; Gu, Jianqiang; Xing, Qirong; Wu, Judy; Zhang, Weili

    2010-11-01

    We demonstrate a superconductor array of subwavelength holes with active thermal control over the resonant transmission induced by surface plasmon polaritons. The array was lithographically fabricated on a high-temperature yttrium barium copper oxide superconductor and characterized by terahertz time-domain spectroscopy. We observe a clear transition from a virtual excitation of the surface plasmon mode to a real surface plasmon mode. The highly controllable superconducting plasmonic crystals may find promising applications in the design of low-loss, large- dynamic-range amplitude modulation and surface-plasmon-based terahertz devices.

  11. Hybrid Arrays for Chemical Sensing

    Science.gov (United States)

    Kramer, Kirsten E.; Rose-Pehrsson, Susan L.; Johnson, Kevin J.; Minor, Christian P.

    In recent years, multisensory approaches to environment monitoring for chemical detection as well as other forms of situational awareness have become increasingly popular. A hybrid sensor is a multimodal system that incorporates several sensing elements and thus produces data that are multivariate in nature and may be significantly increased in complexity compared to data provided by single-sensor systems. Though a hybrid sensor is itself an array, hybrid sensors are often organized into more complex sensing systems through an assortment of network topologies. Part of the reason for the shift to hybrid sensors is due to advancements in sensor technology and computational power available for processing larger amounts of data. There is also ample evidence to support the claim that a multivariate analytical approach is generally superior to univariate measurements because it provides additional redundant and complementary information (Hall, D. L.; Linas, J., Eds., Handbook of Multisensor Data Fusion, CRC, Boca Raton, FL, 2001). However, the benefits of a multisensory approach are not automatically achieved. Interpretation of data from hybrid arrays of sensors requires the analyst to develop an application-specific methodology to optimally fuse the disparate sources of data generated by the hybrid array into useful information characterizing the sample or environment being observed. Consequently, multivariate data analysis techniques such as those employed in the field of chemometrics have become more important in analyzing sensor array data. Depending on the nature of the acquired data, a number of chemometric algorithms may prove useful in the analysis and interpretation of data from hybrid sensor arrays. It is important to note, however, that the challenges posed by the analysis of hybrid sensor array data are not unique to the field of chemical sensing. Applications in electrical and process engineering, remote sensing, medicine, and of course, artificial

  12. Large scale biomimetic membrane arrays

    DEFF Research Database (Denmark)

    Hansen, Jesper Søndergaard; Perry, Mark; Vogel, Jörg

    2009-01-01

    To establish planar biomimetic membranes across large scale partition aperture arrays, we created a disposable single-use horizontal chamber design that supports combined optical-electrical measurements. Functional lipid bilayers could easily and efficiently be established across CO2 laser micro...... peptides and proteins. Next, we tested the scalability of the biomimetic membrane design by establishing lipid bilayers in rectangular 24 x 24 and hexagonal 24 x 27 aperture arrays, respectively. The results presented show that the design is suitable for further developments of sensitive biosensor assays...

  13. Detection of Leishmania infantum in animals and their ectoparasites by conventional PCR and real time PCR.

    Science.gov (United States)

    de Morais, Rayana Carla Silva; Gonçalves, Suênia da Cunha; Costa, Pietra Lemos; da Silva, Kamila Gaudêncio; da Silva, Fernando José; Silva, Rômulo Pessoa E; de Brito, Maria Edileuza Felinto; Brandão-Filho, Sinval Pinto; Dantas-Torres, Filipe; de Paiva-Cavalcanti, Milena

    2013-04-01

    Visceral leishmaniosis (VL) is a parasitic disease caused by Leishmania infantum, which is primarily transmitted by phlebotomine sandflies. However, there has been much speculation on the role of other arthropods in the transmission of VL. Thus, the aim of this study was to assess the presence of L. infantum in cats, dogs and their ectoparasites in a VL-endemic area in northeastern Brazil. DNA was extracted from blood samples and ectoparasites, tested by conventional PCR (cPCR) and quantitative real time PCR (qPCR) targeting the L. infantum kinetoplast DNA. A total of 280 blood samples (from five cats and 275 dogs) and 117 ectoparasites from dogs were collected. Animals were apparently healthy and not previously tested by serological or molecular diagnostic methods. Overall, 213 (76.1 %) animals and 51 (43.6 %) ectoparasites were positive to L. infantum, with mean parasite loads of 795.2, 31.9 and 9.1 fg in dogs, cats and ectoparasites, respectively. Concerning the positivity between dogs and their ectoparasites, 32 (15.3 %) positive dogs were parasitized by positive ectoparasites. The overall concordance between the PCR protocols used was 59.2 %, with qPCR being more efficient than cPCR; 34.1 % of all positive samples were exclusively positive by qPCR. The high number of positive animals and ectoparasites also indicates that they could serve as sentinels or indicators of the circulation of L. infantum in risk areas.

  14. Propidium monoazide reverse transcription PCR and RT-qPCR for detecting infectious enterovirus and norovirus

    Science.gov (United States)

    Presently there is no established cell line or small animal model that allows for the detection of infectious human norovirus. Current methods based on RT-PCR and RT-qPCR detect both infectious and non-infectious virus and thus the conclusions that may be drawn regarding the publ...

  15. Reduction of heteroduplex formation in PCR amplification

    Czech Academy of Sciences Publication Activity Database

    Michu, Elleni; Mráčková, Martina; Vyskot, Boris; Žlůvová, Jitka

    2010-01-01

    Roč. 54, č. 1 (2010), s. 173-176 ISSN 0006-3134 R&D Projects: GA AV ČR(CZ) KJB600040801; GA ČR(CZ) GD204/09/H002; GA AV ČR(CZ) IAA600040801; GA MŠk(CZ) LC06004 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : polymerase chain reaction * reconditioning PCR * mixed-template PCR Subject RIV: BO - Biophysics Impact factor: 1.582, year: 2010

  16. Detection of genetically modified crops by combination of multiplex PCR and low-density DNA microarray.

    Science.gov (United States)

    Zhou, Ping-Ping; Zhang, Jian-Zhong; You, Yuan-Hai; Wu, Yong-Ning

    2008-02-01

    To develop a technique for simultaneous detection of various target genes in Roundup Ready soybean by combining multiplex PCR and low-density DNA microarray. Two sets of the multiplex PCR system were used to amplify the target genes in genetically modified (GM) soybean. Seventeen capture probes (PCR products) and 17 pairs of corresponding primers were designed according to the genetic characteristics of Rroundup Ready soybean (GTS40-3-2), maize (Mon810, Nk603, GA21), canola (T45, MS1/RF1), and rice (SCK) in many identified GM crops. All of the probes were categorized and identified as species-specific probes. One negative probe and one positive control probe were used to assess the efficiency of all reactions, and therefore eliminate any false positive and negative results. After multiplex PCR reaction, amplicons were adulterated with Cy5-dUTP and hybridized with DNA microarray. The array was then scanned to display the specific hybridization signals of target genes. The assay was applied to the analysis of sample of certified transgenic soybean (Roundup Ready GTS40-3-2) and canola (MS1/RF1). A combination technique of multiplex PCR and DNA microarray was successfully developed to identify multi-target genes in Roundup Ready soybean and MS1/RF1 canola with a great specificity and reliability. Reliable identification of genetic characteristics of Roundup Ready of GM soybean from genetically modified crops was achieved at 0.5% transgenic events, indicating a high sensitivity. A combination technique of multiplex PCR and low-density DNA microarray can reliably detect and identify the genetically modified crops.

  17. Genome-wide oligonucleotide-based array comparative genome hybridization analysis of non-isolated congenital diaphragmatic hernia

    NARCIS (Netherlands)

    D.A. Scott; M. Klaassens; A.M. Holder (Ashley); K.P. Lally (Kevin); C.J. Fernandes (Caraciolo); R-J.H. Galjaard (Robert-Jan); D. Tibboel (Dick); J.E.M.M. de Klein (Annelies); B. Lee (Brendan)

    2007-01-01

    textabstractNon-isolated congenital diaphragmatic hernia (CDH+) is a severe birth defect that is often caused by de novo chromosomal anomalies. In this report, we use genome-wide oligonucleotide-based array comparative genome hybridization (aCGH) followed by rapid real-time quantitative PCR analysis

  18. Multiwall carbon nanotube microcavity arrays

    Energy Technology Data Exchange (ETDEWEB)

    Ahmed, Rajib; Butt, Haider, E-mail: h.butt@bham.ac.uk [Nanotechnology Laboratory, School of Mechanical Engineering, University of Birmingham, Birmingham B15 2TT (United Kingdom); Rifat, Ahmmed A. [Integrated Lightwave Research Group, Department of Electrical Engineering, Faculty of Engineering, University of Malaya, Kuala Lumpur 50603 (Malaysia); Yetisen, Ali K.; Yun, Seok Hyun [Harvard Medical School and Wellman Center for Photomedicine, Massachusetts General Hospital, 65 Landsdowne Street, Cambridge, Massachusetts 02139 (United States); Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States); Dai, Qing [National Center for Nanoscience and Technology, Beijing 100190 (China)

    2016-03-21

    Periodic highly dense multi-wall carbon nanotube (MWCNT) arrays can act as photonic materials exhibiting band gaps in the visible regime and beyond terahertz range. MWCNT arrays in square arrangement for nanoscale lattice constants can be configured as a microcavity with predictable resonance frequencies. Here, computational analyses of compact square microcavities (≈0.8 × 0.8 μm{sup 2}) in MWCNT arrays were demonstrated to obtain enhanced quality factors (≈170–180) and narrow-band resonance peaks. Cavity resonances were rationally designed and optimized (nanotube geometry and cavity size) with finite element method. Series (1 × 2 and 1 × 3) and parallel (2 × 1 and 3 × 1) combinations of microcavities were modeled and resonance modes were analyzed. Higher order MWCNT microcavities showed enhanced resonance modes, which were red shifted with increasing Q-factors. Parallel microcavity geometries were also optimized to obtain narrow-band tunable filtering in low-loss communication windows (810, 1336, and 1558 nm). Compact series and parallel MWCNT microcavity arrays may have applications in optical filters and miniaturized optical communication devices.

  19. Gamma-ray array physics

    International Nuclear Information System (INIS)

    Lister, C. J.

    1999-01-01

    In this contribution I am going to discuss the development of large arrays of Compton Suppressed, High Purity Germanium (HpGe) detectors and the physics that has been, that is being, and that will be done with them. These arrays and their science have dominated low-energy nuclear structure research for the last twenty years and will continue to do so in the foreseeable future. John Sharpey Schafer played a visionary role in convincing a skeptical world that the development of these arrays would lead to a path of enlightenment. The extent to which he succeeded can be seen both through the world-wide propagation of ever more sophisticated devices, and through the world-wide propagation of his students. I, personally, would not be working in research if it were not for Johns inspirational leadership. I am eternally grateful to him. Many excellent reviews of array physics have been made in the past which can provide detailed background reading. The review by Paul Nolan, another ex-Sharpey Schafer student, is particularly comprehensive and clear

  20. Photoelectrochemistry of Semiconductor Nanowire Arrays

    Energy Technology Data Exchange (ETDEWEB)

    Mallouk, Thomas E; Redwing, Joan M

    2009-11-10

    This project supported research on the growth and photoelectrochemical characterization of semiconductor nanowire arrays, and on the development of catalytic materials for visible light water splitting to produce hydrogen and oxygen. Silicon nanowires were grown in the pores of anodic aluminum oxide films by the vapor-liquid-solid technique and were characterized electrochemically. Because adventitious doping from the membrane led to high dark currents, silicon nanowire arrays were then grown on silicon substrates. The dependence of the dark current and photovoltage on preparation techniques, wire diameter, and defect density was studied for both p-silicon and p-indium phosphide nanowire arrays. The open circuit photovoltage of liquid junction cells increased with increasing wire diameter, reaching 350 mV for micron-diameter silicon wires. Liquid junction and radial p-n junction solar cells were fabricated from silicon nano- and microwire arrays and tested. Iridium oxide cluster catalysts stabilized by bidentate malonate and succinate ligands were also made and studied for the water oxidation reaction. Highlights of this project included the first papers on silicon and indium phosphide nanowire solar cells, and a new procedure for making ligand-stabilized water oxidation catalysts that can be covalently linked to molecular photosensitizers or electrode surfaces.

  1. Micromolding for ceramic microneedle arrays

    NARCIS (Netherlands)

    van Nieuwkasteele-Bystrova, Svetlana Nikolajevna; Lüttge, Regina

    2011-01-01

    The fabrication process of ceramic microneedle arrays (MNAs) is presented. This includes the manufacturing of an SU-8/Si-master, its double replication resulting in a PDMS mold for production by micromolding and ceramic sintering. The robustness of the replicated structures was tested by means of

  2. LOFAR- The Low Frequency Array

    NARCIS (Netherlands)

    Falcke, H.D.E.

    2006-01-01

    LOFAR is an innovative radio telescope in the frequency range of 10-240 MHz, realized as a phased array. It will become the largest radio telescope in the world in the time frame 2006-2010, located in Northern Europe. LOFAR is being implemented as a Wide Area Sensor Network which connects thousands

  3. Array Technology for Terahertz Imaging

    Science.gov (United States)

    Reck, Theodore; Siles, Jose; Jung, Cecile; Gill, John; Lee, Choonsup; Chattopadhyay, Goutam; Mehdi, Imran; Cooper, Ken

    2012-01-01

    Heterodyne terahertz (0.3 - 3THz) imaging systems are currently limited to single or a low number of pixels. Drastic improvements in imaging sensitivity and speed can be achieved by replacing single pixel systems with an array of detectors. This paper presents an array topology that is being developed at the Jet Propulsion Laboratory based on the micromachining of silicon. This technique fabricates the array's package and waveguide components by plasma etching of silicon, resulting in devices with precision surpassing that of current metal machining techniques. Using silicon increases the versatility of the packaging, enabling a variety of orientations of circuitry within the device which increases circuit density and design options. The design of a two-pixel transceiver utilizing a stacked architecture is presented that achieves a pixel spacing of 10mm. By only allowing coupling from the top and bottom of the package the design can readily be arrayed in two dimensions with a spacing of 10mm x 18mm.

  4. Light weight digital array SAR

    NARCIS (Netherlands)

    Otten, M.; Maas, N.; Bolt, R.; Anitori, L.

    2010-01-01

    A light weight SAR has been designed, suitable for short range tactical UAVs, consisting of a fully digital receive array, and a very compact active transmit antenna. The weight of the complete RF front is expected to be below 3 kg, with a power consumption below 30 W. This X-band system can provide

  5. Solar array manufacturing industry simulation

    Science.gov (United States)

    Chamberlain, R. G.; Firnett, P. J.; Kleine, B.

    1980-01-01

    Solar Array Manufacturing Industry Simulation (SAMIS) program is a standardized model of industry to manufacture silicon solar modules for use in electricity generation. Model is used to develop financial reports that detail requirements, including amounts and prices for materials, labor, facilities, and equipment required by companies.

  6. Directivity of basic linear arrays

    DEFF Research Database (Denmark)

    Bach, Henning

    1970-01-01

    For a linear uniform array ofnelements, an expression is derived for the directivity as a function of the spacing and the phase constants. The cases of isotropic elements, collinear short dipoles, and parallel short dipoles are included. The formula obtained is discussed in some detail and contour...

  7. Solar array flight dynamic experiment

    Science.gov (United States)

    Schock, Richard W.

    1987-01-01

    The purpose of the Solar Array Flight Dynamic Experiment (SAFDE) is to demonstrate the feasibility of on-orbit measurement and ground processing of large space structures' dynamic characteristics. Test definition or verification provides the dynamic characteristic accuracy required for control systems use. An illumination/measurement system was developed to fly on space shuttle flight STS-41D. The system was designed to dynamically evaluate a large solar array called the Solar Array Flight Experiment (SAFE) that had been scheduled for this flight. The SAFDE system consisted of a set of laser diode illuminators, retroreflective targets, an intelligent star tracker receiver and the associated equipment to power, condition, and record the results. In six tests on STS-41D, data was successfully acquired from 18 retroreflector targets and ground processed, post flight, to define the solar array's dynamic characteristic. The flight experiment proved the viability of on-orbit test definition of large space structures dynamic characteristics. Future large space structures controllability should be greatly enhanced by this capability.

  8. PHARUS : PHased ARray Universal SAR

    NARCIS (Netherlands)

    Paquay, M.H.A.; Vermeulen, B.C.B.; Koomen, P.J.; Hoogeboom, P.; Snoeij, P.; Pouwels, H.

    1996-01-01

    In the Netherlands, a polarimetric C-band aircraft SAR (Synthetic Aperture Radar) has been developed. The project is called PHARUS, an acronm for PHased ARray Universal SAR. This instrument serves remote sensing applications. The antenna system contains 48 active modules (expandable to 96). A module

  9. Real-time PCR: Advanced technologies and applications

    Science.gov (United States)

    This book brings together contributions from 20 experts in the field of PCR, providing a broad perspective of the applications of quantitative real-time PCR (qPCR). The editors state in the preface that the aim is to provide detailed insight into underlying principles and methods of qPCR to provide ...

  10. Diagnostic Performance of a Multiplex PCR assay for meningitis in an HIV-infected population in Uganda

    OpenAIRE

    Rhein, Joshua; Bahr, Nathan C; Hemmert, Andrew C; Cloud, Joann L; Bellamkonda, Satya; Oswald, Cody; Lo, Eric; Nabeta, Henry; Kiggundu, Reuben; Akampurira, Andrew; Musubire, Abdu; Williams, Darlisha; Meya, David B; Boulware, David R

    2015-01-01

    Meningitis remains a worldwide problem, and rapid diagnosis is essential to optimize survival. We evaluated the utility of a multiplex PCR test in differentiating possible etiologies of meningitis. Cerebrospinal fluid (CSF) from 69 HIV-infected Ugandan adults with meningitis was collected at diagnosis (n=51) and among persons with cryptococcal meningitis during therapeutic lumbar punctures (n=68). Cryopreserved CSF specimens were analyzed with BioFire FilmArray? Meningitis/Encephalitis panel,...

  11. Aspergillus PCR: one step closer to standardization.

    NARCIS (Netherlands)

    White, P.L.; Bretagne, S.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Schulz, B.; Finnstrom, N.; Mengoli, C.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2010-01-01

    PCR has been used as an aid in the diagnosis of invasive aspergillosis for almost 2 decades. A lack of standardization has limited both its acceptance as a diagnostic tool and multicenter clinical evaluations, preventing its inclusion in disease-defining criteria. In 2006, the European Aspergillus

  12. Polymerase chain reaction (PCR) based molecular characterization ...

    African Journals Online (AJOL)

    Polymerase chain reaction (PCR) based molecular characterization of popular wheat varieties of Khyber Pukhtunkhwa (KPK) region of Pakistan. ... Molecular markers used in this study show high rate of genetic diversity that can be used to assist a breeding program for the improvement of wheat in KPK-Pakistan. Key words: ...

  13. Inverse PCR for Point Mutation Introduction.

    Science.gov (United States)

    Silva, Diogo; Santos, Gustavo; Barroca, Mário; Collins, Tony

    2017-01-01

    Inverse PCR is a powerful tool for the rapid introduction of desired mutations at desired positions in a circular double-stranded DNA sequence. Here, custom-designed mutant primers oriented in the inverse direction are used to amplify the entire circular template with incorporation of the required mutation(s). By careful primer design it can be used to perform such diverse modifications as the introduction of point mutations and multiple mutations, the insertion of new sequences, and even sequence deletions. Three primer formats are commonly used; nonoverlapping, partially overlapping and fully overlapping primers, and here we describe the use of nonoverlapping primers for introduction of a point mutation. Use of such a primer setup in the PCR reaction, with one of the primers containing the desired mismatch mutation, results in the amplification of a linear, double-stranded, mutated product. Methylated template DNA is removed from the nonmethylated PCR product by DpnI digestion and the PCR product is then phosphorylated by polynucleotide kinase treatment before being recircularized by ligation, and transformed to E. coli. This relatively simple site-directed mutagenesis procedure is of major importance in biology and biotechnology today where it is commonly employed for the study and engineering of DNA, RNA, and proteins.

  14. Real-time PCR gene expression profiling

    Czech Academy of Sciences Publication Activity Database

    Kubista, Mikael; Sjögreen, B.; Forootan, A.; Šindelka, Radek; Jonák, Jiří; Andrade, J.M.

    2007-01-01

    Roč. 1, - (2007), s. 56-60 ISSN 1360-8606 R&D Projects: GA AV ČR KJB500520601 Institutional research plan: CEZ:AV0Z50520514 Keywords : real - time PCR, * expression profiling * statistical analysis Subject RIV: EB - Genetics ; Molecular Biology

  15. Polymerase Chain Reaction (PCR) Detection of Schistosoma ...

    African Journals Online (AJOL)

    ... in local creek water. On the known strength of focal effects of environmental conditions, implications of these results in the epidemiology and design of control activities are very encouraging. Keywords: PCR, Cercaria, Schistosomiasis, Oyan Dam, Bulinus Nigerian Journal of Parasitology, Vol. 32 [2] September 2011, pp.

  16. Handheld real-time PCR device.

    Science.gov (United States)

    Ahrberg, Christian D; Ilic, Bojan Robert; Manz, Andreas; Neužil, Pavel

    2016-02-07

    Here we report one of the smallest real-time polymerase chain reaction (PCR) systems to date with an approximate size of 100 mm × 60 mm × 33 mm. The system is an autonomous unit requiring an external 12 V power supply. Four simultaneous reactions are performed in the form of virtual reaction chambers (VRCs) where a ≈200 nL sample is covered with mineral oil and placed on a glass cover slip. Fast, 40 cycle amplification of an amplicon from the H7N9 gene was used to demonstrate the PCR performance. The standard curve slope was -3.02 ± 0.16 cycles at threshold per decade (mean ± standard deviation) corresponding to an amplification efficiency of 0.91 ± 0.05 per cycle (mean ± standard deviation). The PCR device was capable of detecting a single deoxyribonucleic acid (DNA) copy. These results further suggest that our handheld PCR device may have broad, technologically-relevant applications extending to rapid detection of infectious diseases in small clinics.

  17. Microsphere suspension array assays for detection and differentiation of Hendra and Nipah viruses.

    Science.gov (United States)

    Foord, Adam J; White, John R; Colling, Axel; Heine, Hans G

    2013-01-01

    Microsphere suspension array systems enable the simultaneous fluorescent identification of multiple separate nucleotide targets in a single reaction. We have utilized commercially available oligo-tagged microspheres (Luminex MagPlex-TAG) to construct and evaluate multiplexed assays for the detection and differentiation of Hendra virus (HeV) and Nipah virus (NiV). Both these agents are bat-borne zoonotic paramyxoviruses of increasing concern for veterinary and human health. Assays were developed targeting multiple sites within the nucleoprotein (N) and phosphoprotein (P) encoding genes. The relative specificities and sensitivities of the assays were determined using reference isolates of each virus type, samples from experimentally infected horses, and archival veterinary diagnostic submissions. Results were assessed in direct comparison with an established qPCR. The microsphere array assays achieved unequivocal differentiation of HeV and NiV and the sensitivity of HeV detection was comparable to qPCR, indicating high analytical and diagnostic specificity and sensitivity.

  18. Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

    Science.gov (United States)

    Santísima-Trinidad, Ana Belén; Bornay-Llinares, Fernando Jorge; Martín González, Marcos; Pascual Valero, José Antonio; Ros Muñoz, Margarita

    2017-01-01

    The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH) with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO) recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR) and digital PCR (dPCR) were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks. PMID:28377928

  19. Quantitative PCR and Digital PCR for Detection ofAscaris lumbricoidesEggs in Reclaimed Water.

    Science.gov (United States)

    Acosta Soto, Lucrecia; Santísima-Trinidad, Ana Belén; Bornay-Llinares, Fernando Jorge; Martín González, Marcos; Pascual Valero, José Antonio; Ros Muñoz, Margarita

    2017-01-01

    The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH) with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO) recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR) and digital PCR (dPCR) were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.

  20. Quantitative PCR and Digital PCR for Detection of Ascaris lumbricoides Eggs in Reclaimed Water

    Directory of Open Access Journals (Sweden)

    Lucrecia Acosta Soto

    2017-01-01

    Full Text Available The reuse of reclaimed water from wastewater depuration is a widespread and necessary practice in many areas around the world and must be accompanied by adequate and continuous quality control. Ascaris lumbricoides is one of the soil-transmitted helminths (STH with risk for humans due to its high infectivity and an important determinant of transmission is the inadequacy of water supplies and sanitation. The World Health Organization (WHO recommends a limit equal to or lower than one parasitic helminth egg per liter, to reuse reclaimed water for unrestricted irrigation. We present two new protocols of DNA extraction from large volumes of reclaimed water. Quantitative PCR (qPCR and digital PCR (dPCR were able to detect low amounts of A. lumbricoides eggs. By using the first extraction protocol, which processes 500 mL of reclaimed water, qPCR can detect DNA concentrations as low as one A. lumbricoides egg equivalent, while dPCR can detect DNA concentrations as low as five A. lumbricoides egg equivalents. By using the second protocol, which processes 10 L of reclaimed water, qPCR was able to detect DNA concentrations equivalent to 20 A. lumbricoides eggs. This fact indicated the importance of developing new methodologies to detect helminth eggs with higher sensitivity and precision avoiding possible human infection risks.

  1. Real-time PCR (qPCR) primer design using free online software.

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most commonly used fluorescent chemistries are SYBR® Green dyes and TaqMan®, Molecular Beacon or Scorpion probes. SYBR® Green is very simple to use and cost efficient. As SYBR® Green dye binds to any double-stranded DNA product, its success depends greatly on proper primer design. Many types of online primer design software are available, which can be used free of charge to design desirable SYBR® Green-based qPCR primers. This laboratory exercise is intended for those who have a fundamental background in PCR. It addresses the basic fluorescent chemistries of real-time PCR, the basic rules and pitfalls of primer design, and provides a step-by-step protocol for designing SYBR® Green-based primers with free, online software. Copyright © 2010 Wiley Periodicals, Inc.

  2. Digital PCR for direct quantification of viruses without DNA extraction

    OpenAIRE

    Pav?i?, Jernej; ?el, Jana; Milavec, Mojca

    2015-01-01

    DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration mat...

  3. Phased Array of Phased Arrays (PAPA) Laser Systems Architecture

    Science.gov (United States)

    McManamon, P. A. U. L. F.; Thompson, William

    This paper introduces and analyzes revolutionary laser system architecture capable of dramatically reducing the complexity of laser systems while simultaneously increasing capability. The architecture includes three major subsystems. The first is a phased array of laser sources. In this article, we discuss diode-pumped fiber lasers as the elements of the phased array, although other waveguide lasers can also be considered. The second provides wavefront control and electronics beam steering, as described in an IEEE Proceedings article on "Optical Phased Array Technology" [1]. The third is subaperture receiver technology. Combining these three technologies into a new laser systems architecture results in a system that has graceful degradation, can steer to as wide an angle as individual optical phased array subapertures, and can be scaled to high power and large apertures through phasing of a number of subapertures. Diode-pumped fiber lasers are appealing as laser sources because they are electrically pumped, efficient, relatively simple, and scalable to significant power levels (over 100 Watts has been demonstrated from a single diode-pumped fiber laser) [2]. The fiber laser design also lends itself to integration into a phased array. Fiber lasers have been phased. Initial phasing demonstrations have been at low power and were conducted by taking a single source, dividing it into multiple fibers, then phasing them together. To develop this technology further we need to use independent fiber lasers or fiber amplifiers, seeded by a common source, and to increase laser power. As we increase laser power, we will have to learn to cope with nonlinearities in the laser amplifiers. Optical Phased Array technology has demonstrated steering over a 90-degree field of regard [4], although this approach used additional optical components. If we use straightforward optical phased array beam steering without additional optics we can steer with high efficiency to about one-third

  4. Impact of Early Detection of Respiratory Viruses by Multiplex PCR Assay on Clinical Outcomes in Adult Patients.

    Science.gov (United States)

    Rappo, Urania; Schuetz, Audrey N; Jenkins, Stephen G; Calfee, David P; Walsh, Thomas J; Wells, Martin T; Hollenberg, James P; Glesby, Marshall J

    2016-08-01

    Rapid and definitive diagnosis of viral respiratory infections is imperative in patient triage and management. We compared the outcomes for adult patients with positive tests for respiratory viruses at a tertiary care center across two consecutive influenza seasons (winters of 2010-2011 and 2012). Infections were diagnosed by conventional methods in the first season and by multiplex PCR (FilmArray) in the second season. FilmArray decreased the time to diagnosis of influenza compared to conventional methods (median turnaround times of 1.7 h versus 7.7 h, respectively; P = 0.015); FilmArray also decreased the time to diagnosis of non-influenza viruses (1.5 h versus 13.5 h, respectively; P < 0.0001). Multivariate logistic regression found that a diagnosis of influenza by FilmArray was associated with significantly lower odds ratios (ORs) for admission (P = 0.046), length of stay (P = 0.040), duration of antimicrobial use (P = 0.032), and number of chest radiographs (P = 0.005), when controlling for potential confounders. We conclude that the rapid turnaround time, multiplex nature of the test (allowing simultaneous detection of an array of viruses), and superior sensitivity of FilmArray may improve the evaluation and management of patients suspected of having respiratory virus infections. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  5. DNA electrophoresis through microlithographic arrays

    International Nuclear Information System (INIS)

    Sevick, E.M.; Williams, D.R.M.

    1996-01-01

    Electrophoresis is one of the most widely used techniques in biochemistry and genetics for size-separating charged molecular chains such as DNA or synthetic polyelectrolytes. The separation is achieved by driving the chains through a gel with an external electric field. As a result of the field and the obstacles that the medium provides, the chains have different mobilities and are physically separated after a given process time. The macroscopically observed mobility scales inversely with chain size: small molecules move through the medium quickly while larger molecules move more slowly. However, electrophoresis remains a tool that has yet to be optimised for most efficient size separation of polyelectrolytes, particularly large polyelectrolytes, e.g. DNA in excess of 30-50 kbp. Microlithographic arrays etched with an ordered pattern of obstacles provide an attractive alternative to gel media and provide wider avenues for size separation of polyelectrolytes and promote a better understanding of the separation process. Its advantages over gels are (1) the ordered array is durable and can be re-used, (2) the array morphology is ordered and can be standardized for specific separation, and (3) calibration with a marker polyelectrolyte is not required as the array is reproduced to high precision. Most importantly, the array geometry can be graduated along the chip so as to expand the size-dependent regime over larger chain lengths and postpone saturation. In order to predict the effect of obstacles upon the chain-length dependence in mobility and hence, size separation, we study the dynamics of single chains using theory and simulation. We present recent work describing: 1) the release kinetics of a single DNA molecule hooked around a point, frictionless obstacle and in both weak and strong field limits, 2) the mobility of a chain impinging upon point obstacles in an ordered array of obstacles, demonstrating the wide range of interactions possible between the chain and

  6. Large Format Uncooled Focal Plane Array Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Uncooled focal plane arrays have improved dramatically and array sizes of 320x240 elements in a 50-?m pitch are commercially available at affordable cost. Black...

  7. Radiometric Testing of Magnesium Diboride Array (MDA)

    Data.gov (United States)

    National Aeronautics and Space Administration — The objective is to develop a 2-Dimensional Far Infra-Red Magnesium Diboride Array (2D FIR MDA) to use in NASA's future planetary exploration instruments. The array...

  8. Microfabricated Multianalyte Sensor Arrays for Metabolic Monitoring

    National Research Council Canada - National Science Library

    Pishko, Michael V

    2006-01-01

    ...(ethylene glycol) diacrylate or PEG-DA on the array electrodes. The fabricated microarray sensors were individually addressable and with no cross-talk between adjacent array elements as assessed using cyclic voltammetry...

  9. Microfabricated Multianalyte Sensor Arrays for Metabolic Monitoring

    National Research Council Canada - National Science Library

    Pishko, Michael V

    2007-01-01

    ...(ethylene glycol) diacrylate or PEG-DA on the array electrodes. The fabricated microarray sensors were individually addressable and with no cross-talk between adjacent array elements as assessed using cyclic voltammetry...

  10. Leakage analysis of crossbar memristor arrays

    KAUST Repository

    Zidan, Mohammed A.

    2014-07-01

    Crossbar memristor arrays provide a promising high density alternative for the current memory and storage technologies. These arrays suffer from parasitic current components that significantly increase the power consumption, and could ruin the readout operation. In this work we study the trade-off between the crossbar array density and the power consumption required for its readout. Our analysis is based on simulating full memristor arrays on a SPICE platform.

  11. Method to fabricate hollow microneedle arrays

    Energy Technology Data Exchange (ETDEWEB)

    Kravitz, Stanley H [Placitas, NM; Ingersoll, David [Albuquerque, NM; Schmidt, Carrie [Los Lunas, NM; Flemming, Jeb [Albuquerque, NM

    2006-11-07

    An inexpensive and rapid method for fabricating arrays of hollow microneedles uses a photoetchable glass. Furthermore, the glass hollow microneedle array can be used to form a negative mold for replicating microneedles in biocompatible polymers or metals. These microneedle arrays can be used to extract fluids from plants or animals. Glucose transport through these hollow microneedles arrays has been found to be orders of magnitude more rapid than natural diffusion.

  12. Exergetic Optimization of a Solar Photovoltaic Array

    OpenAIRE

    Sarhaddi, Faramarz; Farahat, Said; Ajam, Hossein; Behzadmehr, Amin

    2009-01-01

    An exergetic optimization is developed to determine the optimal performance and design parameters of a solar photovoltaic (PV) array. A detailed energy and exergy analysis is carried out to evaluate the electrical performance, exergy destruction components, and exergy efficiency of a typical PV array. The exergy efficiency of a PV array obtained in this paper is a function of climatic, operating, and design parameters such as ambient temperature, solar radiation intensity, PV array temperatur...

  13. Safety System for a Towed Array

    Science.gov (United States)

    2017-09-25

    300196 1 of 13 SAFETY SYSTEM FOR A TOWED SOURCE STATEMENT OF GOVERNMENT INTEREST [0001] The invention described herein may be manufactured...invention is a towed array safety system and method of use that prevents the loss of a towed array cable and towed array handling system in the event of a...tension surge while retaining required safety features of the towed array handling system . (2) Description of the Prior Art [0004] There have

  14. High voltage load resistor array

    Science.gov (United States)

    Lehmann, Monty Ray [Smithfield, VA

    2005-01-18

    A high voltage resistor comprising an array of a plurality of parallel electrically connected resistor elements each containing a resistive solution, attached at each end thereof to an end plate, and about the circumference of each of the end plates, a corona reduction ring. Each of the resistor elements comprises an insulating tube having an electrode inserted into each end thereof and held in position by one or more hose clamps about the outer periphery of the insulating tube. According to a preferred embodiment, the electrode is fabricated from stainless steel and has a mushroom shape at one end, that inserted into the tube, and a flat end for engagement with the end plates that provides connection of the resistor array and with a load.

  15. Antenna Arrays and Automotive Applications

    CERN Document Server

    Rabinovich, Victor

    2013-01-01

    This book throws a lifeline to designers wading through mounds of antenna array patents looking for the most suitable systems for their projects. Drastically reducing the research time required to locate solutions to the latest challenges in automotive communications, it sorts and systematizes material on cutting-edge antenna arrays that feature multi-element communication systems with enormous potential for the automotive industry. These new systems promise to make driving safer and more efficient, opening up myriad applications, including vehicle-to-vehicle traffic that prevents collisions, automatic toll collection, vehicle location and fine-tuning for cruise control systems. This book’s exhaustive coverage begins with currently deployed systems, frequency ranges and key parameters. It proceeds to examine system geometry, analog and digital beam steering technology (including "smart" beams formed in noisy environments), maximizing signal-to-noise ratios, miniaturization, and base station technology that ...

  16. Invasive tightly coupled processor arrays

    CERN Document Server

    LARI, VAHID

    2016-01-01

    This book introduces new massively parallel computer (MPSoC) architectures called invasive tightly coupled processor arrays. It proposes strategies, architecture designs, and programming interfaces for invasive TCPAs that allow invading and subsequently executing loop programs with strict requirements or guarantees of non-functional execution qualities such as performance, power consumption, and reliability. For the first time, such a configurable processor array architecture consisting of locally interconnected VLIW processing elements can be claimed by programs, either in full or in part, using the principle of invasive computing. Invasive TCPAs provide unprecedented energy efficiency for the parallel execution of nested loop programs by avoiding any global memory access such as GPUs and may even support loops with complex dependencies such as loop-carried dependencies that are not amenable to parallel execution on GPUs. For this purpose, the book proposes different invasion strategies for claiming a desire...

  17. Subwavelength micropillar array terahertz lasers.

    Science.gov (United States)

    Krall, Michael; Brandstetter, Martin; Deutsch, Christoph; Detz, Hermann; Andrews, Aaron Maxwell; Schrenk, Werner; Strasser, Gottfried; Unterrainer, Karl

    2014-01-13

    We report on micropillar-based terahertz lasers with active pillars that are much smaller than the emission wavelength. These micropillar array lasers correspond to scaled-down band-edge photonic crystal lasers forming an active photonic metamaterial. In contrast to photonic crystal lasers which use significantly larger pillar structures, lasing emission is not observed close to high-symmetry points in the photonic band diagram, but in the effective medium regime. We measure stimulated emission at 4 THz for micropillar array lasers with pillar diameters of 5 µm. Our results not only demonstrate the integration of active subwavelength optics in a terahertz laser, but are also an important step towards the realization of nanowire-based terahertz lasers.

  18. Application of oligonucleotide array technology for the rapid detection of pathogenic bacteria of foodborne infections.

    Science.gov (United States)

    Hong, Bang-Xing; Jiang, Li-Fang; Hu, Yu-Shan; Fang, Dan-Yun; Guo, Hui-Yu

    2004-09-01

    A rapid and accurate method for detection for common pathogenic bacteria in foodborne infections was established by using oligonucleotide array technology. Nylon membrane was used as the array support. A mutation region of the 23S rRNA gene was selected as the discrimination target from 14 species (genera) of bacteria causing foodborne infections and two unrelated bacterial species. A pair of universal primers was designed for PCR amplification of the 23S rRNA gene. Twenty-one species (genera)-specific oligonucleotide detection probes were synthesized and spotted onto the nylon membranes. The 23S rRNA gene amplification products of 14 species of pathogenic bacteria were hybridized to the oligonucleotide array. Hybridization results were analyzed with digoxigenin-linked enzyme reaction. Results indicated that nine species of pathogenic bacteria (Escherichia coli, Campylobacter jejuni, Shigella dysenteriae, Vibrio cholerae, Vibrio parahaemolyticus, Proteus vulgaris, Bacillus cereus, Listeria monocytogenes and Clostridium botulinum) showed high sensitivity and specificity for the oligonucleotide array. Two other species (Salmonella enterica and Yersinia enterocolitica) gave weak cross-reaction with E. coli, but the reaction did not affect their detection. After redesigning the probes, positive hybridization results were obtained with Staphylococcus aureus, but not with Clostridium perfringens and Streptococcus pyogenes. The oligonucleotide array can also be applied to samples collected in clinical settings of foodborne infections. The superiority of oligonucleotide array over other tests lies on its rapidity, accuracy and efficiency in the diagnosis, treatment and control of foodborne infections.

  19. Microneedle array electrode for human EEG recording.

    NARCIS (Netherlands)

    Lüttge, Regina; van Nieuwkasteele-Bystrova, Svetlana Nikolajevna; van Putten, Michel Johannes Antonius Maria; Vander Sloten, Jos; Verdonck, Pascal; Nyssen, Marc; Haueisen, Jens

    2009-01-01

    Microneedle array electrodes for EEG significantly reduce the mounting time, particularly by circumvention of the need for skin preparation by scrubbing. We designed a new replication process for numerous types of microneedle arrays. Here, polymer microneedle array electrodes with 64 microneedles,

  20. Ferroelectric transistor memory arrays on flexible foils

    NARCIS (Netherlands)

    Breemen, A. van; Kam, B.; Cobb, B.; Rodriguez, F.G.; Heck, G. van; Myny, K.; Marrani, A.; Vinciguerra, V.; Gelinck, G.H.

    2013-01-01

    In this paper, we successfully fabricated and operated passive matrix P(VDF-TrFE) transistor arrays, i.e. memory arrays in which no pass-transistors or other additional electronic components are used. Because of the smaller cell, a higher integration density is possible. We demonstrate arrays up to

  1. Towards Hybrid Array Types in SAC

    NARCIS (Netherlands)

    Grelck, C.; Tang, F.

    2014-01-01

    Array programming is characterised by a formal calculus of (regular, dense) multidimensional arrays that defines the relationships between structural properties like rank and shape as well as data set sizes. Operations in the array calculus often impose certain constraints on the relationships of

  2. Arrays of magnetic nanoparticles capped with alkylamines

    Indian Academy of Sciences (India)

    While there has been reasonable success in preparing arrays of metal and semiconducting chalcoginide nanoparticles, arrays of metal oxides are relatively unknown. [12,13]. We considered it important to prepare arrays of metal oxide particles with use- ful magnetic properties, particularly in view of their stability under ...

  3. Calibration strategies for the Cherenkov Telescope Array

    NARCIS (Netherlands)

    Gaug, M.; Berge, D.; Daniel, M.; Doro, M.; Förster, A.; Hofmann, W.; Maccarone, M.C.; Parsons, D.; de los Reyes Lopez, R.; van Eldik, C.

    2014-01-01

    The Central Calibration Facilities workpackage of the Cherenkov Telescope Array (CTA) observatory for very high energy gamma ray astronomy defines the overall calibration strategy of the array, develops dedicated hardware and software for the overall array calibration and coordinates the calibration

  4. Array-MAPH: a methodology for the detection of locus copy-number changes in complex genomes.

    Science.gov (United States)

    Kousoulidou, Ludmila; Männik, Katrin; Sismani, Carolina; Zilina, Olga; Parkel, Sven; Puusepp, Helen; Tõnisson, Neeme; Palta, Priit; Remm, Maido; Kurg, Ants; Patsalis, Philippos C

    2008-01-01

    High-throughput genome-wide screening methods to detect subtle genomic imbalances are extremely important for diagnostic genetics and genomics. Here, we provide a detailed protocol for a microarray-based technique, applying the principle of multiplex amplifiable probe hybridization (MAPH). Methodology and software have been developed for designing unique PCR-amplifiable sequences (400-600 bp) covering any genomic region of interest. These sequences are amplified, cloned and spotted onto arrays (targets). A mixture of the same sequences (probes) is hybridized to genomic DNA immobilized on a membrane. Bound probes are recovered and quantitatively amplified by PCR, labeled and hybridized to the array. The procedure can be completed in 4-5 working days, excluding microarray preparation. Unlike array-comparative genomic hybridization (array-CGH), test DNA of specifically reduced complexity is hybridized to an array of identical small amplifiable target sequences, resulting in increased hybridization specificity and higher potential for increasing resolution. Array-MAPH can be used for detection of small-scale copy-number changes in complex genomes, leading to genotype-phenotype correlations and the discovery of new genes.

  5. Si Microwire Array Solar Cells

    Energy Technology Data Exchange (ETDEWEB)

    Putnam, Morgan C.; Boettcher, Shannon W.; Kelzenberg, Michael D.; Turner-Evans, Daniel B.; Spurgeon, Joshua M.; Warren, Emily L.; Briggs, Ryan M.; Lewis, Nathan S.; Atwater, Harry A.

    2010-01-01

    Si microwire-array solar cells with Air Mass 1.5 Global conversion efficiencies of up to 7.9% have been fabricated using an active volume of Si equivalent to a 4 μm thick Si wafer. These solar cells exhibited open-circuit voltages of 500 mV, short-circuit current densities (J{sub sc}) of up to 24 mA cm{sup -2}, and fill factors >65% and employed Al{sub 2}O{sub 3} dielectric particles that scattered light incident in the space between the wires, a Ag back reflector that prevented the escape of incident illumination from the back surface of the solar cell, and an a-SiN{sub x}:H passivation/anti-reflection layer. Wire-array solar cells without some or all of these design features were also fabricated to demonstrate the importance of the light-trapping elements in achieving a high J{sub sc}. Scanning photocurrent microscopy images of the microwire-array solar cells revealed that the higher J{sub sc} of the most advanced cell design resulted from an increased absorption of light incident in the space between the wires. Spectral response measurements further revealed that solar cells with light-trapping elements exhibited improved red and infrared response, as compared to solar cells without light-trapping elements.

  6. LOFAR, the low frequency array

    Science.gov (United States)

    Vermeulen, R. C.

    2012-09-01

    LOFAR, the Low Frequency Array, is a next-generation radio telescope designed by ASTRON, with antenna stations concentrated in the north of the Netherlands and currently spread into Germany, France, Sweden and the United Kingdom; plans for more LOFAR stations exist in several other countries. Utilizing a novel, phased-array design, LOFAR is optimized for the largely unexplored low frequency range between 30 and 240 MHz. Digital beam-forming techniques make the LOFAR system agile and allow for rapid re-pointing of the telescopes as well as the potential for multiple simultaneous observations. Processing (e.g. cross-correlation) takes place in the LOFAR BlueGene/P supercomputer, and associated post-processing facilities. With its dense core (inner few km) array and long (more than 1000 km) interferometric baselines, LOFAR reaches unparalleled sensitivity and resolution in the low frequency radio regime. The International LOFAR Telescope (ILT) is now issuing its first call for observing projects that will be peer reviewed and selected for observing starting in December. Part of the allocations will be made on the basis of a fully Open Skies policy; there are also reserved fractions assigned by national consortia in return for contributions from their country to the ILT. In this invited talk, the gradually expanding complement of operationally verified observing modes and capabilities are reviewed, and some of the exciting first astronomical results are presented.

  7. Evaluation of low density array technology for quantitative parallel measurement of multiple genes in human tissue

    Directory of Open Access Journals (Sweden)

    Harmer Daniel W

    2006-02-01

    Full Text Available Abstract Background Low density arrays (LDAs have recently been introduced as a novel approach to gene expression profiling. Based on real time quantitative RT-PCR (QRT-PCR, these arrays enable a more focused and sensitive approach to the study of gene expression than gene chips, while offering higher throughput than more established approaches to QRT-PCR. We have now evaluated LDAs as a means of determining the expression of multiple genes simultaneously in human tissues and cells. Results Comparisons between LDAs reveal low variability, with correlation coefficients close to 1. By performing 2-fold and 10-fold serial dilutions of cDNA samples in the LDAs we determined a clear linear relationship between the gene expression data points over 5 orders of magnitude. We also showed that it is possible to use LDAs to accurately and quantitatively detect 2-fold changes in target copy number as well as measuring genes that are expressed with low and high copy numbers in the range of 1 × 102 – 1 × 106 copies. Furthermore, the data generated by the LDA from a cell based pharmacological study were comparable to data generated by conventional QRT-PCR. Conclusion LDAs represent a valuable new approach for sensitive and quantitative gene expression profiling.

  8. Application of droplet digital PCR for quantitative detection of Spiroplasma citri in comparison with real time PCR.

    Directory of Open Access Journals (Sweden)

    Yogita Maheshwari

    Full Text Available Droplet digital polymerase chain reaction (ddPCR is a method for performing digital PCR that is based on water-oil emulsion droplet technology. It is a unique approach to measure the absolute copy number of nucleic acid targets without the need of external standards. This study evaluated the applicability of ddPCR as a quantitative detection tool for the Spiroplasma citri, causal agent of citrus stubborn disease (CSD in citrus. Two sets of primers, SP1, based on the spiral in housekeeping gene, and a multicopy prophage gene, SpV1 ORF1, were used to evaluate ddPCR in comparison with real time (quantitative PCR (qPCR for S. citri detection in citrus tissues. Standard curve analyses on tenfold dilution series showed that both ddPCR and qPCR exhibited good linearity and efficiency. However, ddPCR had a tenfold greater sensitivity than qPCR and accurately quantified up to one copy of spiralin gene. Receiver operating characteristic analysis indicated that the ddPCR methodology was more robust for diagnosis of CSD and the area under the curve was significantly broader compared to qPCR. Field samples were used to validate ddPCR efficacy and demonstrated that it was equal or better than qPCR to detect S. citri infection in fruit columella due to a higher pathogen titer. The ddPCR assay detected both the S. citri spiralin and the SpV1 ORF1 targets quantitatively with high precision and accuracy compared to qPCR assay. The ddPCR was highly reproducible and repeatable for both the targets and showed higher resilience to PCR inhibitors in citrus tissue extract for the quantification of S. citri compare to qPCR.

  9. Clostridium perfringens isolate typing by multiplex PCR

    Directory of Open Access Journals (Sweden)

    MR Ahsani

    2010-01-01

    Full Text Available Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.

  10. The Colorado Lightning Mapping Array

    Science.gov (United States)

    Rison, W.; Krehbiel, P. R.; Thomas, R. J.; Rodeheffer, D.; Fuchs, B.

    2012-12-01

    A fifteen station Lightning Mapping Array (LMA) was installed in northern Colorado in the spring of 2012. While the driving force for the array was to produce 3-dimensional lightning data to support the Deep Convective Clouds and Chemistry (DC3) Experiment (Barth, this conference), data from the array are being used for several other projects. These include: electrification studies in conjunction with the CSU CHILL radar (Lang et al, this conference); observations of the parent lightning discharges of sprites (Lyons et al, this conference); trying to detect upward discharges triggered by wind turbines, characterizing conditions in which aircraft flying through clouds produce discharges which can be detected by the LMA, and other opportunities, such as observations of lightning in pyrocumulus clouds produced by the High Park Fire west of Fort Collins, CO. All the COLMA stations are solar-powered, and use broadband cellular modems for data communications. This makes the stations completely self-contained and autonomous, allowing a station to be installed anywhere a cellular signal is available. Because most of the stations were installed well away from anthropogenic noise sources, the COLMA is very sensitive. This is evidenced by the numerous plane tracks detected in its the vicinity. The diameter, D, of the COLMA is about 100 km, significantly larger than other LMAs. Because the error in the radial distance r is proportional to (r/D)2, and the error in the altitude z is proportional to (z/D)2, the larger array diameter greatly expands the usable range of the COLMA. The COLMA is able to detect and characterize lighting flashes to a distance of about 350 km from the array center. In addition to a web-based display (lightning.nmt.edu/colma), geo-referenced images are produced and updated at one-minute intervals. These geo-referenced images can be used to overlay the real-time lightning data on Google Earth and other mapping software. These displays were used by the DC3

  11. Acoustic array systems theory, implementation, and application

    CERN Document Server

    Bai, Mingsian R; Benesty, Jacob

    2013-01-01

    Presents a unified framework of far-field and near-field array techniques for noise source identification and sound field visualization, from theory to application. Acoustic Array Systems: Theory, Implementation, and Application provides an overview of microphone array technology with applications in noise source identification and sound field visualization. In the comprehensive treatment of microphone arrays, the topics covered include an introduction to the theory, far-field and near-field array signal processing algorithms, practical implementations, and common applic

  12. Pneumocystis PCR: It Is Time to Make PCR the Test of Choice

    Science.gov (United States)

    Doyle, Laura; Vogel, Sherilynn

    2017-01-01

    Abstract Background The testing strategy for Pneumocystis at the Cleveland Clinic changed from toluidine blue staining to polymerase chain reaction (PCR). We studied the differences in positivity rates for these assays and compared each with the detection of Pneumocystis in companion specimens by cytology and surgical pathology. Methods We reviewed the results of all Pneumocystis test orders 1 year before and 1 year after the implementation of a Pneumocystis-specific PCR. We also reviewed the corresponding cytology and surgical pathology results, if performed. Finally, we reviewed the medical records of patients with rare Pneumocystis detected by PCR in an effort to differentiate colonization vs true disease. Results Toluidine blue staining and surgical pathology had similar sensitivities and negative predictive values, both of which were superior to cytology. There was a >4-fold increase in the annual detection of Pneumocystis by PCR compared with toluidine blue staining (toluidine blue staining: 11/1583 [0.69%] vs PCR: 44/1457 [3.0%]; chi-square P < .001). PCR detected 1 more case than surgical pathology and was far more sensitive than cytology. Chart review demonstrated that the vast majority of patients with rare Pneumocystis detected were immunosuppressed, had radiologic findings supportive of this infection, had no other pathogens detected, and were treated for pneumocystosis by the clinical team. Conclusion PCR was the most sensitive method for the detection of Pneumocystis and should be considered the diagnostic test of choice. Correlation with clinical and radiologic findings affords discrimination of early true disease from the far rarer instances of colonization. PMID:29062861

  13. Influence of PCR reagents on DNA polymerase extension rates measured on real-time PCR instruments.

    Science.gov (United States)

    Montgomery, Jesse L; Wittwer, Carl T

    2014-02-01

    Radioactive DNA polymerase activity methods are cumbersome and do not provide initial extension rates. A simple extension rate assay would enable study of basic assumptions about PCR and define the limits of rapid PCR. A continuous assay that monitors DNA polymerase extension using noncovalent DNA dyes on common real-time PCR instruments was developed. Extension rates were measured in nucleotides per second per molecule of polymerase. To initiate the reaction, a nucleotide analog was heat activated at 95 °C for 5 min, the temperature decreased to 75 °C, and fluorescence monitored until substrate exhaustion in 30-90 min. The assay was linear with time for over 40% of the reaction and for polymerase concentrations over a 100-fold range (1-100 pmol/L). Extension rates decreased continuously with increasing monovalent cation concentrations (lithium, sodium, potassium, cesium, and ammonium). Melting-temperature depressors had variable effects. DMSO increased rates up to 33%, whereas glycerol had little effect. Betaine, formamide, and 1,2-propanediol decreased rates with increasing concentrations. Four common noncovalent DNA dyes inhibited polymerase extension. Heat-activated nucleotide analogs were 92% activated after 5 min, and hot start DNA polymerases were 73%-90% activated after 20 min. Simple DNA extension rate assays can be performed on real-time PCR instruments. Activity is decreased by monovalent cations, DNA dyes, and most melting temperature depressors. Rational inclusion of PCR components on the basis of their effects on polymerase extension is likely to be useful in PCR, particularly rapid-cycle or fast PCR.

  14. Status of wake and array loss research

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, D.L.

    1991-09-01

    In recent years, many projects have evaluated wind turbine wake effects and resultant array losses in both Europe and the United States. This paper examines the status of current knowledge about wake effects and array losses and suggests future research. Single-turbine wake characteristics have been studied extensively and are generally described well by existing theoretical models. Field measurements of wake effects in wind turbine arrays are largely limited to small arrays, with 2 to 4 rows of turbines. Few data have been published on wake effects within large arrays. Measurements of wake deficits downwind of large arrays that deficits are substantially larger and extend farther downwind than expected. Although array design models have been developed, these models have been tested and verified using only limited data from a few rows of wind turbines in complex terrain, whereas some of the largest arrays have more than 40 rows of wind turbines. Planned cooperative efforts with the wind industry will obtain existing data relevant to analyzing energy deficits within large arrays and identifying data sets for potential use in array model verification efforts. Future research being considered include a cooperative research experiment to obtain more definitive data on wake deficits and turbulence within and downwind of large arrays. 16 refs., 9 figs., 1 tab.

  15. Coupling Between Waveguide-Fed Slot Arrays

    Science.gov (United States)

    Rengarajan, Sembiam

    2011-01-01

    Coupling between two waveguide-fed planar slot arrays has been investigated using full-wave analysis. The analysis employs the method-of-moments solution to the pertinent coupled integral equations for the aperture electric field of all slots. In order to compute coupling between two arrays, the input port of the first array is excited with a TE(sub 10) mode wave while the second one is match-terminated. After solving the moment method matrix equations, the aperture fields of all slots are obtained and thereby the TE(sub 10) mode wave received at the input port of the second array is determined. Coupling between two arrays is the ratio of the wave amplitude arriving in the second array port to the incident wave amplitude at the first array port. The coupling mechanism has been studied as a function of spacing between arrays in different directions, e.g. the electric field plane, the magnetic field plane, and the diagonal plane. Computed coupling values are presented for different array geometries. This work is novel since it provides a good understanding of coupling between waveguide-fed slot arrays as a function of spacing and orientation for different aperture distributions and array architectures. This serves as a useful tool for antenna design engineers and system engineers.

  16. Design of circular differential microphone arrays

    CERN Document Server

    Benesty, Jacob; Cohen, Israel

    2015-01-01

    Recently, we proposed a completely novel and efficient way to design differential beamforming algorithms for linear microphone arrays. Thanks to this very flexible approach, any order of differential arrays can be designed. Moreover, they can be made robust against white noise amplification, which is the main inconvenience in these types of arrays. The other well-known problem with linear arrays is that electronic steering is not really feasible.  In this book, we extend all these fundamental ideas to circular microphone arrays and show that we can design small and compact differential arrays of any order that can be electronically steered in many different directions and offer a good degree of control of the white noise amplification problem, high directional gain, and frequency-independent response. We also present a number of practical examples, demonstrating that differential beamforming with circular microphone arrays is likely one of the best candidates for applications involving speech enhancement (i....

  17. Reducing fiber cross-talk in mineral fiber arrays

    OpenAIRE

    Daniel Lee Stark

    2017-01-01

    Monocentric optics replace current systems with diffraction limited performance. The fiber arrays have been the issue. Commercial expensive fiber arrays are available, but enhanced mineral fiber arrays offer very inexpensive fiber arrays.

  18. Microbial diversity in a thermophilic aerobic biofilm process: analysis by length heterogeneity PCR (LH-PCR).

    Science.gov (United States)

    Tiirola, Marja A; Suvilampi, Juhani E; Kulomaa, Markku S; Rintala, Jukka A

    2003-05-01

    A two-stage pilot-scale thermophilic aerobic suspended carrier biofilm process (SCBP) was set up for the on-site treatment of pulp and paper mill whitewater lining. The microbial diversity in this process was analyzed by length heterogeneity analysis of PCR-amplified 16S ribosomal DNA. The primer pair selected for PCR amplification was first evaluated by a computational analysis of fragment lengths in ten main phylogenetical eubacterial groups. The fragment contained the first third of the 16S rRNA gene, which was shown to vary naturally between 465 and 563 bp in length. The length heterogeneity analysis of polymerase chain reaction (LH-PCR) profile of the biomass attached to carrier elements was found to be diverse in both stages of the SCBP. During normal operating conditions, sequences belonging to beta-Proteobacteria, Cytophaga/Flexibacter/Bacteroides group and gamma-Proteobacteria were assigned to the most prominent LH-PCR peak. Samples from the suspended biomass consisted of completely different bacterial populations, which were, however, similar in the serial reactors. The pilot process experienced alkaline shocks, after which Bacillus-like sequences were detected in both the biofilm and suspended biomass. However, when the conditions were reversed, the normal microbial population in the biofilm recovered rapidly without further biomass inoculations. This study shows that LH-PCR is a valuable method for profiling microbial diversity and dynamics in industrial wastewater processes.

  19. Lab-on-a-chip PCR: real time PCR in miniaturized format for HLA diagnostics

    Science.gov (United States)

    Gaertner, Claudia; Becker, Holger; Hlawatsch, Nadine; Klemm, Richard; Moche, Christian; Sewart, René; Frank, Rainer; Willems, Andreas

    2014-05-01

    In case of transplantation or the identification of special metabolic diseases like coeliac disease, HLA typing has to be done fast and reliably with easy-to-handle devices by using limited amount of sample. Against this background a lab-on-a-chip device was realized enabling a fast HLA typing via miniaturized Real-time PCR. Hereby, two main process steps were combined, namely the extraction of DNA from whole blood and the amplification of the target DNA by Real-time PCR giving rise-to a semi-quantitative analysis. For the implementation of both processes on chip, a sample preparation and a real-time module were used. Sample preparation was carried out by using magnetic beads that were stored directly on chip as dry powder, together with all lysis reagents. After purification of the DNA by applying a special buffer regime, the sample DNA was transferred into the PCR module for amplification and detection. Coping with a massively increased surface-to-volume ratio, which results in a higher amount of unspecific binding on the chip surface, special additives needed to be integrated to compensate for this effect. Finally the overall procedure showed a sensitivity comparable to standard Real-time PCR but reduced the duration of analysis to significantly less than one hour. The presented work demonstrates that the combination of lab-on-a-chip PCR with direct optical read-out in a real-time fashion is an extremely promising tool for molecular diagnostics.

  20. Species identification and quantification in meat and meat products using droplet digital PCR (ddPCR).

    Science.gov (United States)

    Floren, C; Wiedemann, I; Brenig, B; Schütz, E; Beck, J

    2015-04-15

    Species fraud and product mislabelling in processed food, albeit not being a direct health issue, often results in consumer distrust. Therefore methods for quantification of undeclared species are needed. Targeting mitochondrial DNA, e.g. CYTB gene, for species quantification is unsuitable, due to a fivefold inter-tissue variation in mtDNA content per cell resulting in either an under- (-70%) or overestimation (+160%) of species DNA contents. Here, we describe a reliable two-step droplet digital PCR (ddPCR) assay targeting the nuclear F2 gene for precise quantification of cattle, horse, and pig in processed meat products. The ddPCR assay is advantageous over qPCR showing a limit of quantification (LOQ) and detection (LOD) in different meat products of 0.01% and 0.001%, respectively. The specificity was verified in 14 different species. Hence, determining F2 in food by ddPCR can be recommended for quality assurance and control in production systems. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. Real-Time Polymerase Chain Reaction (PCR) Capability in Space

    Data.gov (United States)

    National Aeronautics and Space Administration — The goal of this project is enabling the real-time polymerase chain reaction (real-time PCR) technology in space. In space, the real-time PCR technology can be used...

  2. The FPGA Pixel Array Detector

    International Nuclear Information System (INIS)

    Hromalik, Marianne S.; Green, Katherine S.; Philipp, Hugh T.; Tate, Mark W.; Gruner, Sol M.

    2013-01-01

    A proposed design for a reconfigurable x-ray Pixel Array Detector (PAD) is described. It operates by integrating a high-end commercial field programmable gate array (FPGA) into a 3-layer device along with a high-resistivity diode detection layer and a custom, application-specific integrated circuit (ASIC) layer. The ASIC layer contains an energy-discriminating photon-counting front end with photon hits streamed directly to the FPGA via a massively parallel, high-speed data connection. FPGA resources can be allocated to perform user defined tasks on the pixel data streams, including the implementation of a direct time autocorrelation function (ACF) with time resolution down to 100 ns. Using the FPGA at the front end to calculate the ACF reduces the required data transfer rate by several orders of magnitude when compared to a fast framing detector. The FPGA-ASIC high-speed interface, as well as the in-FPGA implementation of a real-time ACF for x-ray photon correlation spectroscopy experiments has been designed and simulated. A 16×16 pixel prototype of the ASIC has been fabricated and is being tested. -- Highlights: ► We describe the novelty and need for the FPGA Pixel Array Detector. ► We describe the specifications and design of the Diode, ASIC and FPGA layers. ► We highlight the Autocorrelation Function (ACF) for speckle as an example application. ► Simulated FPGA output calculates the ACF for different input bitstreams to 100 ns. ► Reduced data transfer rate by 640× and sped up real-time ACF by 100× other methods.

  3. Space-Based Radar Array System Simulation and Validation.

    Science.gov (United States)

    1980-09-01

    external field RBCA (NBFA,3), Array of position vectors R pointing to the center of RBC(NBF,3) ’ array a or array b element current segments from array...to center of array a RBCA (NBFA,3) element current segments from array a element feedpoint TSXB, TSYB Array b phase factors YDEL Skew parameter for...Number of feedback modes used in mutual coupling calcu- lation (POLX, POLY, POLZ) Polarization vector of external field RBCA (NBFA,3), Array of position

  4. rasdaman Array Database: current status

    Science.gov (United States)

    Merticariu, George; Toader, Alexandru

    2015-04-01

    rasdaman (Raster Data Manager) is a Free Open Source Array Database Management System which provides functionality for storing and processing massive amounts of raster data in the form of multidimensional arrays. The user can access, process and delete the data using SQL. The key features of rasdaman are: flexibility (datasets of any dimensionality can be processed with the help of SQL queries), scalability (rasdaman's distributed architecture enables it to seamlessly run on cloud infrastructures while offering an increase in performance with the increase of computation resources), performance (real-time access, processing, mixing and filtering of arrays of any dimensionality) and reliability (legacy communication protocol replaced with a new one based on cutting edge technology - Google Protocol Buffers and ZeroMQ). Among the data with which the system works, we can count 1D time series, 2D remote sensing imagery, 3D image time series, 3D geophysical data, and 4D atmospheric and climate data. Most of these representations cannot be stored only in the form of raw arrays, as the location information of the contents is also important for having a correct geoposition on Earth. This is defined by ISO 19123 as coverage data. rasdaman provides coverage data support through the Petascope service. Extensions were added on top of rasdaman in order to provide support for the Geoscience community. The following OGC standards are currently supported: Web Map Service (WMS), Web Coverage Service (WCS), and Web Coverage Processing Service (WCPS). The Web Map Service is an extension which provides zoom and pan navigation over images provided by a map server. Starting with version 9.1, rasdaman supports WMS version 1.3. The Web Coverage Service provides capabilities for downloading multi-dimensional coverage data. Support is also provided for several extensions of this service: Subsetting Extension, Scaling Extension, and, starting with version 9.1, Transaction Extension, which

  5. Conformal Antenna Array Design Handbook

    Science.gov (United States)

    1981-09-01

    PLOTI-e2#Yo2) AsYSCL*(J-1 )-YSL CALL N~IE(7.*L ,-LH2tH~oto4HF4.0) GO TO 10 11 CALL PLOT(-oltY92) 10 YuY ..Y DX. XM/NX NNX=NX+1 DO 20 K-19NNX CALL PLUT...polynomial. Then the coefficients are determined from the Chebyshev polynomial (Taylor, 1952; Tseng and Cheng , 1968). As in the case of linear arrays, a...used ( Cheng and Tseng, 1967). Difference pattern parameters such as slope (Powers, 1967) or directivity (Bayliss, 1968) can also be maximized at the

  6. Coated carbon nanotube array electrodes

    Science.gov (United States)

    Ren, Zhifeng [Newton, MA; Wen, Jian [Newton, MA; Chen, Jinghua [Chestnut Hill, MA; Huang, Zhongping [Belmont, MA; Wang, Dezhi [Wellesley, MA

    2008-10-28

    The present invention provides conductive carbon nanotube (CNT) electrode materials comprising aligned CNT substrates coated with an electrically conducting polymer, and the fabrication of electrodes for use in high performance electrical energy storage devices. In particular, the present invention provides conductive CNTs electrode material whose electrical properties render them especially suitable for use in high efficiency rechargeable batteries. The present invention also provides methods for obtaining surface modified conductive CNT electrode materials comprising an array of individual linear, aligned CNTs having a uniform surface coating of an electrically conductive polymer such as polypyrrole, and their use in electrical energy storage devices.

  7. Rapid detection of dermatophytes and Candida albicans in onychomycosis specimens by an oligonucleotide array.

    Science.gov (United States)

    Han, Huan Wen; Hsu, Mark Ming-Long; Choi, Jong Soo; Hsu, Chao-Kai; Hsieh, Hsin Yi; Li, Hsin Chieh; Chang, Hsien Chang; Chang, Tsung Chain

    2014-11-07

    Onychomycosis is a fungal infection of nails, leading to the gradual destruction of the nail plate. Treatment of onychomycosis may need long-time oral antifungal therapy that can have potential side effects, thus accurate diagnosis of the disease before treatment is important. Culture for diagnosis of onychomycosis is time-consuming and has high false-negative rates. To expedite the diagnosis, an oligonucleotide array, based on hybridization between immobilized oligonucleotide probes and PCR products, for direct detection of dermatophytes and Candida albicans in clinical specimens was evaluated. Species-specific oligonucleotide probes designed from the internal transcribed spacer (ITS) regions of the rRNA gene were immobilized on a nylon membrane. The assay procedures consisted of PCR amplification of the ITS using universal primers, followed by hybridization of the digoxigenin-labeled amplicons to probes on the array. Thirty two nail samples (29 patients) were analyzed by the array, and the results were compared with those obtained by culture. Array-positive but culture-negative samples were confirmed by cloning and re-sequencing of the amplified ITS and by reviewing patient's clinical data. The total recovery of culture and confirmed array-positive but culture-negative results was considered 100% and was used for performance evaluation of both methods. Concordant results were obtained in 21 samples (10 positives and 11 negatives) by both methods. Eleven samples were array-positive but culture-negative; among them, 9 samples were considered true positives after discrepant analysis. Comparing with culture, the array had significantly higher sensitivity [100% (95% CI 82.2% -100%) vs 52.6% (28.9% -75.5%), p <0.001] and negative predictive value [100% (71.3% -100%) vs 59.1% (36.4% -79.3%), p <0.05), while no significant differences were observed in specificity (84.6% vs 100%, p =0.48) and positive predictive value (90.5% vs 100%, p =1.0). The whole procedures of the

  8. Development of -Specific Real-Time PCR Assays for the Detection and Identification of Eight Medically Important Species

    Directory of Open Access Journals (Sweden)

    Jing Zhang

    2016-01-01

    Full Text Available Culture-based identification methods have been the gold standard for the diagnosis of fungal infection. Currently, molecular technologies such as real-time PCR assays with short turnaround time can provide desirable alternatives for the rapid detection of Candida microbes. However, most of the published PCR primer sets are not Candida specific and likely to amplify DNA from common environmental contaminants, such as Aspergillus microbes. In this study, we designed pan- Candida primer sets based on the ribosomal DNA-coding regions conserved within Candida but distinct from those of Aspergillus and Penicillium. We demonstrate that the final two selected pan- Candida primer sets would not amplify Aspergillus DNA and could be used to differentiate eight medically important Candida pathogens in real-time PCR assays based on their melting profiles, with a sensitivity of detection as low as 10 fg of Candida genomic DNA. Moreover, we further evaluated and selected species-specific primer sets covering Candida albicans, Candida glabrata, Candida tropicalis , and Candida dubliniensis and show that they had high sensitivity and specificity. These real-time PCR primer sets could potentially be assembled into a single PCR array for the rapid detection of Candida species in various clinical settings, such as corneal transplantation.

  9. Analysis of extracellular RNA by digital PCR

    Directory of Open Access Journals (Sweden)

    Kenji eTakahashi

    2014-06-01

    Full Text Available The transfer of extracellular RNA is emerging as an important mechanism for intracellular communication. The ability for the transfer of functionally active RNA molecules from one cell to another within vesicles such as exosomes enables a cell to modulate cellular signaling and biological processes within recipient cells. The study of extracellular RNA requires sensitive methods for the detection of these molecules. In this methods article, we will describe protocols for the detection of such extracellular RNA using sensitive detection technologies such as digital PCR. These protocols should be valuable to researchers interested in the role and contribution of extracellular RNA to tumor cell biology.

  10. Layout Of Antennas And Cables In A Large Array

    Science.gov (United States)

    Logan, Ronald T., Jr.

    1995-01-01

    Layout devised to minimize total land area occupied by large phased array of antennas and to minimize total length of cables in array. In original intended application, array expanded version of array of paraboloidal-dish microwave communication antennas of Deep Space Network. Layout also advantageous for other phased arrays of antennas and antenna elements, including notably printed-circuit microwave antenna arrays.

  11. Biomarker discovery for colon cancer using a 761 gene RT-PCR assay

    Directory of Open Access Journals (Sweden)

    Hackett James R

    2007-08-01

    Full Text Available Abstract Background Reverse transcription PCR (RT-PCR is widely recognized to be the gold standard method for quantifying gene expression. Studies using RT-PCR technology as a discovery tool have historically been limited to relatively small gene sets compared to other gene expression platforms such as microarrays. We have recently shown that TaqMan® RT-PCR can be scaled up to profile expression for 192 genes in fixed paraffin-embedded (FPE clinical study tumor specimens. This technology has also been used to develop and commercialize a widely used clinical test for breast cancer prognosis and prediction, the Onco typeDX™ assay. A similar need exists in colon cancer for a test that provides information on the likelihood of disease recurrence in colon cancer (prognosis and the likelihood of tumor response to standard chemotherapy regimens (prediction. We have now scaled our RT-PCR assay to efficiently screen 761 biomarkers across hundreds of patient samples and applied this process to biomarker discovery in colon cancer. This screening strategy remains attractive due to the inherent advantages of maintaining platform consistency from discovery through clinical application. Results RNA was extracted from formalin fixed paraffin embedded (FPE tissue, as old as 28 years, from 354 patients enrolled in NSABP C-01 and C-02 colon cancer studies. Multiplexed reverse transcription reactions were performed using a gene specific primer pool containing 761 unique primers. PCR was performed as independent TaqMan® reactions for each candidate gene. Hierarchal clustering demonstrates that genes expected to co-express form obvious, distinct and in certain cases very tightly correlated clusters, validating the reliability of this technical approach to biomarker discovery. Conclusion We have developed a high throughput, quantitatively precise multi-analyte gene expression platform for biomarker discovery that approaches low density DNA arrays in numbers of

  12. Determining Fungi rRNA Copy Number by PCR

    Science.gov (United States)

    The goal of this project is to improve the quantification of indoor fungal pollutants via the specific application of quantitative PCR (qPCR). Improvement will be made in the controls used in current qPCR applications. This work focuses on the use of two separate controls within ...

  13. Evaluation of Aspergillus PCR Protocols for Testing Serum Specimens

    NARCIS (Netherlands)

    White, P.L.; Mengoli, C.; Bretagne, S.; Cuenca-Estrella, M.; Finnstrom, N.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2011-01-01

    A panel of human serum samples spiked with various amounts of Aspergillus fumigatus genomic DNA was distributed to 23 centers within the European Aspergillus PCR Initiative to determine analytical performance of PCR. Information regarding specific methodological components and PCR performance was

  14. Real-Time PCR for Universal Phytoplasma Detection and Quantification

    DEFF Research Database (Denmark)

    Christensen, Nynne Meyn; Nyskjold, Henriette; Nicolaisen, Mogens

    2013-01-01

    Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs and in other...

  15. (PCR) for direct cloning of blunt-end DNA fragments

    African Journals Online (AJOL)

    Administrator

    2011-09-19

    Sep 19, 2011 ... interest by PCR using proof reading DNA polymerase, such as Pfu, KOD and Primerstar, is preferred since the. PCR products with a higher degree of fidelity are required in many investigations. However, traditional blunt-end cloning method for direct cloning of blunt-end PCR products is not efficient since ...

  16. Selecting PCR for the Diagnosis of Intestinal Parasitosis

    DEFF Research Database (Denmark)

    Hartmeyer, G. N.; Hoegh, S. V.; Skov, M. N.

    2017-01-01

    , Cryptosporidium species, or Entamoeba histolytica detected by microscopy. We evaluated and selected in-house singleplex real-time PCR (RT-PCR) assays for these pathogens in 99 stool samples from patients suspected of having intestinal parasitosis tested by microscopy. The strategy included a genus-specific PCR...

  17. ArrayBridge: Interweaving declarative array processing with high-performance computing

    Energy Technology Data Exchange (ETDEWEB)

    Xing, Haoyuan [The Ohio State Univ., Columbus, OH (United States); Floratos, Sofoklis [The Ohio State Univ., Columbus, OH (United States); Blanas, Spyros [The Ohio State Univ., Columbus, OH (United States); Byna, Suren [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Prabhat, Prabhat [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Wu, Kesheng [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Brown, Paul [Paradigm4, Inc., Waltham, MA (United States)

    2017-05-04

    Scientists are increasingly turning to datacenter-scale computers to produce and analyze massive arrays. Despite decades of database research that extols the virtues of declarative query processing, scientists still write, debug and parallelize imperative HPC kernels even for the most mundane queries. This impedance mismatch has been partly attributed to the cumbersome data loading process; in response, the database community has proposed in situ mechanisms to access data in scientific file formats. Scientists, however, desire more than a passive access method that reads arrays from files. This paper describes ArrayBridge, a bi-directional array view mechanism for scientific file formats, that aims to make declarative array manipulations interoperable with imperative file-centric analyses. Our prototype implementation of ArrayBridge uses HDF5 as the underlying array storage library and seamlessly integrates into the SciDB open-source array database system. In addition to fast querying over external array objects, ArrayBridge produces arrays in the HDF5 file format just as easily as it can read from it. ArrayBridge also supports time travel queries from imperative kernels through the unmodified HDF5 API, and automatically deduplicates between array versions for space efficiency. Our extensive performance evaluation in NERSC, a large-scale scientific computing facility, shows that ArrayBridge exhibits statistically indistinguishable performance and I/O scalability to the native SciDB storage engine.

  18. Comparison of a Commercially Available Repetitive-Element PCR System (DiversiLab) with PCR Ribotyping for Typing of Clostridium difficile Strains ▿

    OpenAIRE

    Eckert, C.; Van Broeck, J.; Spigaglia, P.; Burghoffer, B.; Delmée, M.; Mastrantonio, P.; Barbut, F.

    2011-01-01

    This study compared a repetitive-element PCR (rep-PCR) method (DiversiLab system) to PCR ribotyping. The discriminatory power of rep-PCR was 0.997. Among the PCR ribotype 027 isolates tested, different rep types could be distinguished. rep-PCR showed a higher discriminatory power than PCR ribotyping. Nevertheless, this method requires technical skill, and visual interpretation of rep-PCR fingerprint patterns may be difficult.

  19. An efficient multistrategy DNA decontamination procedure of PCR reagents for hypersensitive PCR applications.

    Directory of Open Access Journals (Sweden)

    Sophie Champlot

    Full Text Available BACKGROUND: PCR amplification of minute quantities of degraded DNA for ancient DNA research, forensic analyses, wildlife studies and ultrasensitive diagnostics is often hampered by contamination problems. The extent of these problems is inversely related to DNA concentration and target fragment size and concern (i sample contamination, (ii laboratory surface contamination, (iii carry-over contamination, and (iv contamination of reagents. METHODOLOGY/PRINCIPAL FINDINGS: Here we performed a quantitative evaluation of current decontamination methods for these last three sources of contamination, and developed a new procedure to eliminate contaminating DNA contained in PCR reagents. We observed that most current decontamination methods are either not efficient enough to degrade short contaminating DNA molecules, rendered inefficient by the reagents themselves, or interfere with the PCR when used at doses high enough to eliminate these molecules. We also show that efficient reagent decontamination can be achieved by using a combination of treatments adapted to different reagent categories. Our procedure involves γ- and UV-irradiation and treatment with a mutant recombinant heat-labile double-strand specific DNase from the Antarctic shrimp Pandalus borealis. Optimal performance of these treatments is achieved in narrow experimental conditions that have been precisely analyzed and defined herein. CONCLUSIONS/SIGNIFICANCE: There is not a single decontamination method valid for all possible contamination sources occurring in PCR reagents and in the molecular biology laboratory and most common decontamination methods are not efficient enough to decontaminate short DNA fragments of low concentration. We developed a versatile multistrategy decontamination procedure for PCR reagents. We demonstrate that this procedure allows efficient reagent decontamination while preserving the efficiency of PCR amplification of minute quantities of DNA.

  20. Micromirror arrays for object selection

    Science.gov (United States)

    Waldis, Severin; Zamkotsian, Frederic; Clerc, Pierre-Andre; Zickar, Michael; Noell, Wilfried; de Rooij, Nico

    2007-10-01

    We report on micromirror arrays being developed for object selection in Multi Object Spectrographs for astronomical applications. The micromirrors are etched in bulk single crystal silicon whereas the cantilever type suspension is realized by surface micromachining. One micromirror element is 100μm × 200μm in size. The micromirrors are actuated electrostatically by electrodes located on a second chip. The use of silicon on insulator (SOI) wafers for both mirror and electrode chip ensures thermal compatibility for cryogenic operation. A system of multiple landing beams has been developed, which passively locks the mirror at a well defined tilt angle when actuated. The mechanical tilt angle obtained is 20° at a pull-in voltage of 90V. Measurements with an optical profiler showed that the tilt angle of the actuated and locked mirror is stable with a precision of one arc minute over a range of 15V. This locking system makes the tilt angle merely independent from process variations across the wafer and thus provides uniform tilt angle over the whole array. The precision on tilt angle from mirror to mirror measured is one arc minute. The surface quality of the mirrors in actuated state is better than 10nm peak-to-valley and the local roughness is around 1nm RMS. Preliminary cryogenic tests showed that the micromirror device sustains 120K without any structural damage.

  1. Sensitivity of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood

    OpenAIRE

    da Costa Lima, Manoel Sebastião; Zorzenon, Denielly Christina Rodrigues; Dorval, Maria Elizabeth Cavalheiros; Pontes, Elenir Rose Jardim Cury; Oshiro, Elisa Teruya; Cunha, Rodrigo; Andreotti, Renato; Matos, Maria de Fatima Cepa

    2013-01-01

    Objective: To evaluate the effectiveness of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood samples. Methods: DNA extraction was performed using Promega Wizard襅 Genomic kits. PCR employing RV1/RV2 primers yielded 1 45-bp amplicons. Real-time PCR was performed with the same primers and SYBR Green ROX Plus mix. These techniques were used to analyze 100 peripheral blood samples from patients with clinical signs of the disease. Results...

  2. Identification of periodontopathogen microorganisms by PCR technique

    Directory of Open Access Journals (Sweden)

    Milićević Radovan

    2008-01-01

    Full Text Available INTRODUCTION Periodontitis is an inflammatory disease of the supporting tissues of teeth and is a major cause of tooth loss in adults. The onset and progression of periodontal disease is attributed to the presence of elevated levels of a consortium of pathogenic bacteria. Gram negative bacteria, mainly strict anaerobes, play the major role. OBJECTIVE The present study aimed to assess the presence of the main types of microorganisms involved in the aetiopathogenesis of periodontal disease: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Eikenella corrodens, Treponema denticola, Tanerella forsythia and Prevotella intermedia in different samples collected from the oral cavity of 90 patients diagnosed with periodontitis. METHOD Bacterial DNA detection was performed in diverse biological materials, namely in dental plaque, gingival tissue and saliva, by means of multiplex PCR, a technique that allows simultaneous identification of two different bacterial genomes. RESULTS In the dental plaque of the periodontitis patients, Treponema denticola dominated. In the gingival tissue, Tannerella forsythia and Treponema denticola were the microbiota most frequently detected, whilst in saliva Treponema denticola and Eikenella corrodens were found with the highest percentage. CONCLUSION The identification of microorganisms by multiplex PCR is specific and sensitive. Rapid and precise assessment of different types of periodontopathogens is extremely important for early detection of the infection and consequently for the prevention and treatment of periodontal disease. In everyday clinical practice, for routine bacterial evaluation in patients with periodontal disease, the dental plaque is the most suitable biological material, because it is the richest in periodontal bacteria.

  3. Heterozygote PCR product melting curve prediction.

    Science.gov (United States)

    Dwight, Zachary L; Palais, Robert; Kent, Jana; Wittwer, Carl T

    2014-03-01

    Melting curve prediction of PCR products is limited to perfectly complementary strands. Multiple domains are calculated by recursive nearest neighbor thermodynamics. However, the melting curve of an amplicon containing a heterozygous single-nucleotide variant (SNV) after PCR is the composite of four duplexes: two matched homoduplexes and two mismatched heteroduplexes. To better predict the shape of composite heterozygote melting curves, 52 experimental curves were compared with brute force in silico predictions varying two parameters simultaneously: the relative contribution of heteroduplex products and an ionic scaling factor for mismatched tetrads. Heteroduplex products contributed 25.7 ± 6.7% to the composite melting curve, varying from 23%-28% for different SNV classes. The effect of ions on mismatch tetrads scaled to 76%-96% of normal (depending on SNV class) and averaged 88 ± 16.4%. Based on uMelt (www.dna.utah.edu/umelt/umelt.html) with an expanded nearest neighbor thermodynamic set that includes mismatched base pairs, uMelt HETS calculates helicity as a function of temperature for homoduplex and heteroduplex products, as well as the composite curve expected from heterozygotes. It is an interactive Web tool for efficient genotyping design, heterozygote melting curve prediction, and quality control of melting curve experiments. The application was developed in Actionscript and can be found online at http://www.dna.utah.edu/hets/. © 2013 WILEY PERIODICALS, INC.

  4. Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?

    Science.gov (United States)

    Morton, C Oliver; White, P Lewis; Barnes, Rosemary A; Klingspor, Lena; Cuenca-Estrella, Manuel; Lagrou, Katrien; Bretagne, Stéphane; Melchers, Willem; Mengoli, Carlo; Caliendo, Angela M; Cogliati, Massimo; Debets-Ossenkopp, Yvette; Gorton, Rebecca; Hagen, Ferry; Halliday, Catriona; Hamal, Petr; Harvey-Wood, Kathleen; Jaton, Katia; Johnson, Gemma; Kidd, Sarah; Lengerova, Martina; Lass-Florl, Cornelia; Linton, Chris; Millon, Laurence; Morrissey, C Orla; Paholcsek, Melinda; Talento, Alida Fe; Ruhnke, Markus; Willinger, Birgit; Donnelly, J Peter; Loeffler, Juergen

    2017-06-01

    A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. Light Trapping with Silicon Light Funnel Arrays.

    Science.gov (United States)

    Prajapati, Ashish; Nissan, Yuval; Gabay, Tamir; Shalev, Gil

    2018-03-19

    Silicon light funnels are three-dimensional subwavelength structures in the shape of inverted cones with respect to the incoming illumination. Light funnel (LF) arrays can serve as efficient absorbing layers on account of their light trapping capabilities, which are associated with the presence of high-density complex Mie modes. Specifically, light funnel arrays exhibit broadband absorption enhancement of the solar spectrum. In the current study, we numerically explore the optical coupling between surface light funnel arrays and the underlying substrates. We show that the absorption in the LF array-substrate complex is higher than the absorption in LF arrays of the same height (~10% increase). This, we suggest, implies that a LF array serves as an efficient surface element that imparts additional momentum components to the impinging illumination, and hence optically excites the substrate by near-field light concentration, excitation of traveling guided modes in the substrate, and mode hybridization.

  6. Light Trapping with Silicon Light Funnel Arrays

    Directory of Open Access Journals (Sweden)

    Ashish Prajapati

    2018-03-01

    Full Text Available Silicon light funnels are three-dimensional subwavelength structures in the shape of inverted cones with respect to the incoming illumination. Light funnel (LF arrays can serve as efficient absorbing layers on account of their light trapping capabilities, which are associated with the presence of high-density complex Mie modes. Specifically, light funnel arrays exhibit broadband absorption enhancement of the solar spectrum. In the current study, we numerically explore the optical coupling between surface light funnel arrays and the underlying substrates. We show that the absorption in the LF array-substrate complex is higher than the absorption in LF arrays of the same height (~10% increase. This, we suggest, implies that a LF array serves as an efficient surface element that imparts additional momentum components to the impinging illumination, and hence optically excites the substrate by near-field light concentration, excitation of traveling guided modes in the substrate, and mode hybridization.

  7. Photovoltaic array space power plus diagnostics experiment

    Science.gov (United States)

    Guidice, Donald A.

    1990-01-01

    The objective of the Photovoltaic Array Space Power Plus Diagnostics (PASP Plus) experiment is to measure the effects of the interaction of the low- to mid-altitude space environment on the performance of a diverse set of small solar-cell arrays (planar and concentrator, representative of present and future military technologies) under differing conditions of velocity-vector orientation and simulated (by biasing) high-voltage operation. Solar arrays to be tested include Si and GaAs planar arrays and several types of GaAs concentrator arrays. Diagnostics (a Langmuir probe and a pressure gauge) and a transient pulse monitor (to measure radiated and conducted EMI during arcing) will be used to determine the impact of the environment on array operation to help verify various interactions models. Results from a successful PASP Plus flight will furnish answers to important interactions questions and provide inputs for design and test standards for photovoltaic space-power subsystems.

  8. Space and power efficient hybrid counters array

    Science.gov (United States)

    Gara, Alan G [Mount Kisco, NY; Salapura, Valentina [Chappaqua, NY

    2009-05-12

    A hybrid counter array device for counting events. The hybrid counter array includes a first counter portion comprising N counter devices, each counter device for receiving signals representing occurrences of events from an event source and providing a first count value corresponding to a lower order bits of the hybrid counter array. The hybrid counter array includes a second counter portion comprising a memory array device having N addressable memory locations in correspondence with the N counter devices, each addressable memory location for storing a second count value representing higher order bits of the hybrid counter array. A control device monitors each of the N counter devices of the first counter portion and initiates updating a value of a corresponding second count value stored at the corresponding addressable memory location in the second counter portion. Thus, a combination of the first and second count values provide an instantaneous measure of number of events received.

  9. Generating Milton Babbitt's all-partition arrays

    DEFF Research Database (Denmark)

    Bemman, Brian; Meredith, David

    2016-01-01

    In most of Milton Babbitt's (1916–2011) works written since the early 1960s, both the pitch and rhythmic content is organized according to a highly constrained structure known as the all-partition array. The all-partition array provides a framework that ensures that as many different forms...... backtracking algorithm for generating a particular type of all-partition array found in Babbitt’s works, known as a Smalley array. Constructing such an array is a difficult task, and we present two heuristics for helping to generate this type of structure. We provide the parameter values required...... by this algorithm to generate the specific all-partition arrays used in three of Babbitt’s works. Finally, we evaluate the algorithm and the heuristics in terms of how well they predict the sequences of integer partitions used in two of Babbitt’s works. We also explore the effect of the heuristics...

  10. Mapping Electrical Crosstalk in Pixelated Sensor Arrays

    Science.gov (United States)

    Seshadri, Suresh (Inventor); Cole, David (Inventor); Smith, Roger M. (Inventor); Hancock, Bruce R. (Inventor)

    2017-01-01

    The effects of inter pixel capacitance in a pixilated array may be measured by first resetting all pixels in the array to a first voltage, where a first image is read out, followed by resetting only a subset of pixels in the array to a second voltage, where a second image is read out, where the difference in the first and second images provide information about the inter pixel capacitance. Other embodiments are described and claimed.

  11. Halbach arrays in precision motion control

    Energy Technology Data Exchange (ETDEWEB)

    Trumper, D.L.; Williams, M.E. [Massachusetts Institute of Technology, Cambridge, MA (United States)

    1995-02-01

    The Halbach array was developed for use as an optical element in particle accelerators. Following up on a suggestion from Klaus Halbach, the authors have investigated the utility of such arrays as the permanent magnet structure for synchronous machines in cartesian, polar, and cylindrical geometries. Their work has focused on the design of a novel Halbach array linear motor for use in a magnetic suspension stage for photolithography. This paper presents the details of the motor design and its force and power characteristics.

  12. Big Data Challenges for Large Radio Arrays

    Science.gov (United States)

    Jones, Dayton L.; Wagstaff, Kiri; Thompson, David; D'Addario, Larry; Navarro, Robert; Mattmann, Chris; Majid, Walid; Lazio, Joseph; Preston, Robert; Rebbapragada, Umaa

    2012-01-01

    Future large radio astronomy arrays, particularly the Square Kilometre Array (SKA), will be able to generate data at rates far higher than can be analyzed or stored affordably with current practices. This is, by definition, a "big data" problem, and requires an end-to-end solution if future radio arrays are to reach their full scientific potential. Similar data processing, transport, storage, and management challenges face next-generation facilities in many other fields.

  13. Multi-Channel Capacitive Sensor Arrays

    OpenAIRE

    Wang, Bingnan; Long, Jiang; Teo, Koon Hoo

    2016-01-01

    In this paper, multi-channel capacitive sensor arrays based on microstrip band-stop filters are studied. The sensor arrays can be used to detect the proximity of objects at different positions and directions. Each capacitive sensing structure in the array is connected to an inductive element to form resonance at different frequencies. The resonances are designed to be isolated in the frequency spectrum, such that the change in one channel does not affect resonances at other channels. The indu...

  14. Magnetic remanence of Josephson junction arrays

    OpenAIRE

    Passos, W. A. C.; Araujo-Moreira, F. M.; Ortiz, W. A.

    1999-01-01

    In this work we study the magnetic remanence exhibited by Josephson junction arrays in response to an excitation with an AC magnetic field. The effect, predicted by numerical simulations to occur in a range of temperatures, is clearly seen in our tridimensional disordered arrays. We also discuss the influence of the critical current distribution on the temperature interval within which the array develops a magnetic remanence. This effect can be used to determine the critical current distribut...

  15. Report for Development of a Census Array and Evaluation of the Array to Detect Biothreat Agents and Environmental Samples for DHS

    Energy Technology Data Exchange (ETDEWEB)

    Jaing, C; Jackson, P

    2011-04-14

    The objective of this project is to provide DHS a comprehensive evaluation of the current genomic technologies including genotyping, Taqman PCR, multiple locus variable tandem repeat analysis (MLVA), microarray and high-throughput DNA sequencing in the analysis of biothreat agents from complex environmental samples. This report focuses on the design, testing and results of samples on the Census Array. We designed a Census/Detection Array to detect all sequenced viruses (including phage), bacteria (eubacteria), and plasmids. Family-specific probes were selected for all sequenced viral and bacterial complete genomes, segments, and plasmids. Probes were designed to tolerate some sequence variation to enable detection of divergent species with homology to sequenced organisms, and to be unique relative to the human genome. A combination of 'detection' probes with high levels of conservation within a family plus 'census' probes targeting strain/isolate specific regions enabled detection and taxonomic classification from the level of family down to the strain. The array has wider coverage of bacterial and viral targets based on more recent sequence data and more probes per target than other microbial detection/discovery arrays in the literature. We tested the array with purified bacterial and viral DNA/RNA samples, artificial mixes of known bacterial/viral samples, spiked DNA against complex background including BW aerosol samples and soil samples, and environmental samples to evaluate the array's sensitivity and forensic capability. The data were analyzed using our novel maximum likelihood software. For most of the organisms tested, we have achieved at least species level discrimination.

  16. Detection of pseudorabies virus by duplex droplet digital PCR assay.

    Science.gov (United States)

    Ren, Meishen; Lin, Hua; Chen, Shijie; Yang, Miao; An, Wei; Wang, Yin; Xue, Changhua; Sun, Yinjie; Yan, Yubao; Hu, Juan

    2018-01-01

    Aujeszky's disease, caused by pseudorabies virus (PRV), has damaged the economy of the Chinese swine industry. A large number of PRV gene-deleted vaccines have been constructed based on deletion of the glycoprotein E ( gE) gene combined with other virulence-related gene deletions, such as thymidine kinase ( TK), whereas PRV wild-type strains contain an intact gE gene. We developed a sensitive duplex droplet digital PCR (ddPCR) assay to rapidly detect PRV wild-type isolates and gE gene-deleted viral vaccines. We compared this assay with a TaqMan real-time PCR (qPCR) using the same primers and probes. Both assays exhibited good linearity and repeatability; however, ddPCR maintained linearity at extremely low concentrations, whereas qPCR did not. Based on positive results for both gE and gB, the detection limit of ddPCR was found to be 4.75 copies/µL in contrast of 76 copies/µL for qPCR, showing that ddPCR provided a 16-fold improvement in sensitivity. In addition, no nonspecific amplification was shown in specificity testing, and the PRV wild-type was distinguished from a gE-deleted strain. The ddPCR was more sensitive when analyzing clinical serum samples. Thus, ddPCR may become an appropriate detection platform for PRV.

  17. Accurate quantification of supercoiled DNA by digital PCR

    Science.gov (United States)

    Dong, Lianhua; Yoo, Hee-Bong; Wang, Jing; Park, Sang-Ryoul

    2016-01-01

    Digital PCR (dPCR) as an enumeration-based quantification method is capable of quantifying the DNA copy number without the help of standards. However, it can generate false results when the PCR conditions are not optimized. A recent international comparison (CCQM P154) showed that most laboratories significantly underestimated the concentration of supercoiled plasmid DNA by dPCR. Mostly, supercoiled DNAs are linearized before dPCR to avoid such underestimations. The present study was conducted to overcome this problem. In the bilateral comparison, the National Institute of Metrology, China (NIM) optimized and applied dPCR for supercoiled DNA determination, whereas Korea Research Institute of Standards and Science (KRISS) prepared the unknown samples and quantified them by flow cytometry. In this study, several factors like selection of the PCR master mix, the fluorescent label, and the position of the primers were evaluated for quantifying supercoiled DNA by dPCR. This work confirmed that a 16S PCR master mix avoided poor amplification of the supercoiled DNA, whereas HEX labels on dPCR probe resulted in robust amplification curves. Optimizing the dPCR assay based on these two observations resulted in accurate quantification of supercoiled DNA without preanalytical linearization. This result was validated in close agreement (101~113%) with the result from flow cytometry. PMID:27063649

  18. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR)

    Science.gov (United States)

    Felder, Eva; Mossbrugger, Ilona; Lange, Mirko; Wölfel, Roman

    2012-01-01

    Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR) assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix. PMID:23105972

  19. PCR and real-time PCR assays to detect fungi of Alternaria alternata species.

    Science.gov (United States)

    Kordalewska, Milena; Brillowska-Dąbrowska, Anna; Jagielski, Tomasz; Dworecka-Kaszak, Bożena

    2015-01-01

    Fungi of the Alternaria genus are mostly associated with allergic diseases. However, with a growing number of immunocompromised patients, these fungi, with A. alternata being the most prevalent one, are increasingly recognized as etiological agents of infections (phaeohyphomycoses) in humans. Nowadays, identification of Alternaria spp. requires their pure culture and is solely based on morphological criteria. Clinically, Alternaria infections may be indistinguishable from other fungal diseases. Therefore, a diagnostic result is often delayed or even not achieved at all. In this paper we present easy to perform and interpret PCR and real-time PCR assays enabling detection of A. alternata species. On the basis of alignment of β-tubulin gene sequences, A. alternata-specific primers were designed. DNA from fungal isolates, extracted in a two-step procedure, were used in PCR and real-time PCR assays followed by electrophoresis or melting temperature analysis, respectively. The assays specificity was confirmed, since positive results were obtained for all A. alternata isolates, and no positive results were obtained neither for other molds, dermatophytes, yeast-like fungi, nor human DNA. The assays developed here enable fast and unambiguous identification of A. alternata pathogens.

  20. cDNA amplification by SMART-PCR and suppression subtractive hybridization (SSH)-PCR.

    Science.gov (United States)

    Hillmann, Andrew; Dunne, Eimear; Kenny, Dermot

    2009-01-01

    The comparison of two RNA populations that differ from the effects of a single-independent variable, such as a drug treatment or a specific genetic defect, can identify differences in the abundance of specific transcripts that vary in a population-dependent manner. There are a variety of methods for identifying differentially expressed genes, including microarray, SAGE, qRT-PCR, and DDGE. This protocol describes a potentially less sensitive yet relatively easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under investigation and is particularly applicable when minimal levels of starting material, RNA, are available. RNA input can often be a limiting factor when analyzing RNA from, for example, rigorously purified blood cells. This protocol describes the use of SMART-PCR to amplify cDNA from sub-microgram levels of RNA. The amplified cDNA populations under comparison are then subjected to suppression subtractive hybridization (SSH-PCR), a technique that couples subtractive hybridization with suppression PCR to selectively amplify fragments of differentially expressed genes. The final products are cDNA populations enriched for significantly over-represented transcripts in either of the two input RNA preparations. These cDNA populations may then be cloned to make subtracted cDNA libraries and/or used as probes to screen subtracted cDNA, global cDNA, or genomic DNA libraries.

  1. Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR

    Directory of Open Access Journals (Sweden)

    Roman Wölfel

    2012-08-01

    Full Text Available Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.

  2. Processes and Materials for Flexible PV Arrays

    National Research Council Canada - National Science Library

    Gierow, Paul

    2002-01-01

    .... A parallel incentive for development of flexible PV arrays are the possibilities of synergistic advantages for certain types of spacecraft, in particular the Solar Thermal Propulsion (STP) Vehicle...

  3. Large Format Uncooled Focal Plane Array Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Black Forest Engineering has identified innovative modifications in uncooled focal plane array (UFPA) architecture and processing that allows development of large...

  4. Analytical Performance of Four Polymerase Chain Reaction (PCR) and Real Time PCR (qPCR) Assays for the Detection of Six Leishmania Species DNA in Colombia

    Science.gov (United States)

    León, Cielo M.; Muñoz, Marina; Hernández, Carolina; Ayala, Martha S.; Flórez, Carolina; Teherán, Aníbal; Cubides, Juan R.; Ramírez, Juan D.

    2017-01-01

    Leishmaniasis comprises a spectrum of parasitic diseases caused by protozoans of the genus Leishmania. Molecular tools have been widely employed for the detection of Leishmania due to its high sensitivity and specificity. However, the analytical performance of molecular platforms as PCR and real time PCR (qPCR) including a wide variety of molecular markers has never been evaluated. Herein, the aim was to evaluate the analytical performance of 4 PCR-based assays (designed on four different targets) and applied on conventional and real-time PCR platforms. We evaluated the analytical performance of conventional PCR and real time PCR, determining exclusivity and inclusivity, Anticipated Reportable Range (ARR), limit of detection (LoD) and accuracy using primers directed to kDNA, HSP70, 18S and ITS-1 targets. We observed that the kDNA was the most sensitive but does not meet the criterion of exclusivity. The HSP70 presented a higher LoD in conventional PCR and qPCR in comparison with the other markers (1 × 101 and 1 × 10-1 equivalent parasites/mL respectively) and had a higher coefficient of variation in qPCR. No statistically significant differences were found between the days of the test with the four molecular markers. The present study revealed that the 18S marker presented the best performance in terms of analytical sensitivity and specificity for the qPCR in the species tested (species circulating in Colombia). Therefore, we recommend to explore the analytical and diagnostic performance in future studies using a broader number of species across America. PMID:29046670

  5. Improved PCR performance using template DNA from formalin-fixed and paraffin-embedded tissues by overcoming PCR inhibition.

    Directory of Open Access Journals (Sweden)

    Dimo Dietrich

    Full Text Available Formalin-fixed and paraffin-embedded (FFPE tissues represent a valuable source for biomarker studies and clinical routine diagnostics. However, they suffer from degradation of nucleic acids due to the fixation process. Since genetic and epigenetic studies usually require PCR amplification, this degradation hampers its use significantly, impairing PCR robustness or necessitating short amplicons. In routine laboratory medicine a highly robust PCR performance is mandatory for the clinical utility of genetic and epigenetic biomarkers. Therefore, methods to improve PCR performance using DNA from FFPE tissue are highly desired and of wider interest. The effect of template DNA derived from FFPE tissues on PCR performance was investigated by means of qPCR and conventional PCR using PCR fragments of different sizes. DNA fragmentation was analyzed via agarose gel electrophoresis. This study showed that poor PCR amplification was partly caused by inhibition of the DNA polymerase by fragmented DNA from FFPE tissue and not only due to the absence of intact template molecules of sufficient integrity. This PCR inhibition was successfully minimized by increasing the polymerase concentration, dNTP concentration and PCR elongation time thereby allowing for the robust amplification of larger amplicons. This was shown for genomic template DNA as well as for bisulfite-converted template DNA required for DNA methylation analyses. In conclusion, PCR using DNA from FFPE tissue suffers from inhibition which can be alleviated by adaptation of the PCR conditions, therefore allowing for a significant improvement of PCR performance with regard to variability and the generation of larger amplicons. The presented solutions to overcome this PCR inhibition are of tremendous value for clinical chemistry and laboratory medicine.

  6. A Laboratory-Developed TaqMan Array Card for Simultaneous Detection of 19 Enteropathogens

    Science.gov (United States)

    Liu, Jie; Gratz, Jean; Amour, Caroline; Kibiki, Gibson; Becker, Stephen; Janaki, Lalitha; Verweij, Jaco J.; Taniuchi, Mami; Sobuz, Shihab U.; Haque, Rashidul; Haverstick, Doris M.

    2013-01-01

    The TaqMan Array Card (TAC) system is a 384-well singleplex real-time PCR format that has been used to detect multiple infection targets. Here we developed an enteric TaqMan Array Card to detect 19 enteropathogens, including viruses (adenovirus, astrovirus, norovirus GII, rotavirus, and sapovirus), bacteria (Campylobacter jejuni/C. coli, Clostridium difficile, Salmonella, Vibrio cholerae, diarrheagenic Escherichia coli strains including enteroaggregative E. coli [EAEC], enterotoxigenic E. coli [ETEC], enteropathogenic E. coli [EPEC], and Shiga-toxigenic E. coli [STEC]), Shigella/enteroinvasive E. coli (EIEC), protozoa (Cryptosporidium, Giardia lamblia, and Entamoeba histolytica), and helminths (Ascaris lumbricoides and Trichuris trichiura), as well as two extrinsic controls to monitor extraction and amplification efficiency (the bacteriophage MS2 and phocine herpesvirus). Primers and probes were newly designed or adapted from published sources and spotted onto microfluidic cards. Fecal samples were spiked with extrinsic controls, and DNA and RNA were extracted using the QiaAmp Stool DNA minikit and the QuickGene RNA Tissue kit, respectively, and then mixed with Ag-Path-ID One Step real-time reverse transcription-PCR (RT-PCR) reagents and loaded into cards. PCR efficiencies were between 90% and 105%, with linearities of 0.988 to 1. The limit of detection of the assays in the TAC was within a 10-fold difference from the cognate assays performed on plates. Precision testing demonstrated a coefficient of variation of below 5% within a run and 14% between runs. Accuracy was evaluated for 109 selected clinical specimens and revealed an average sensitivity and specificity of 85% and 77%, respectively, compared with conventional methods (including microscopy, culture, and immunoassay) and 98% and 96%, respectively, compared with our laboratory-developed PCR-Luminex assays. This TAC allows fast, accurate, and quantitative detection of a broad spectrum of enteropathogens and

  7. Molecular quantification of environmental DNA using microfluidics and digital PCR.

    Science.gov (United States)

    Hoshino, Tatsuhiko; Inagaki, Fumio

    2012-09-01

    Real-time PCR has been widely used to evaluate gene abundance in natural microbial habitats. However, PCR-inhibitory substances often reduce the efficiency of PCR, leading to the underestimation of target gene copy numbers. Digital PCR using microfluidics is a new approach that allows absolute quantification of DNA molecules. In this study, digital PCR was applied to environmental samples, and the effect of PCR inhibitors on DNA quantification was tested. In the control experiment using λ DNA and humic acids, underestimation of λ DNA at 1/4400 of the theoretical value was observed with 6.58 ng μL(-1) humic acids. In contrast, digital PCR provided accurate quantification data with a concentration of humic acids up to 9.34 ng μL(-1). The inhibitory effect of paddy field soil extract on quantification of the archaeal 16S rRNA gene was also tested. By diluting the DNA extract, quantified copy numbers from real-time PCR and digital PCR became similar, indicating that dilution was a useful way to remedy PCR inhibition. The dilution strategy was, however, not applicable to all natural environmental samples. For example, when marine subsurface sediment samples were tested the copy number of archaeal 16S rRNA genes was 1.04×10(3) copies/g-sediment by digital PCR, whereas real-time PCR only resulted in 4.64×10(2) copies/g-sediment, which was most likely due to an inhibitory effect. The data from this study demonstrated that inhibitory substances had little effect on DNA quantification using microfluidics and digital PCR, and showed the great advantages of digital PCR in accurate quantifications of DNA extracted from various microbial habitats. Copyright © 2012 Elsevier GmbH. All rights reserved.

  8. Molecular identification of common Salmonella serovars using multiplex DNA sensor-based suspension array.

    Science.gov (United States)

    Aydin, Muhsin; Carter-Conger, Jacqueline; Gao, Ning; Gilmore, David F; Ricke, Steven C; Ahn, Soohyoun

    2018-04-01

    Salmonella is one of major foodborne pathogens and the leading cause of foodborne illness-related hospitalizations and deaths. It is critical to develop a sensitive and rapid detection assay that can identify Salmonella to ensure food safety. In this study, a DNA sensor-based suspension array system of high multiplexing ability was developed to identify eight Salmonella serovars commonly associated with foodborne outbreaks to the serotype level. Each DNA sensor was prepared by activating pre-encoded microspheres with oligonucleotide probes that are targeting virulence genes and serovar-specific regions. The mixture of 12 different types of DNA sensors were loaded into a 96-well microplate and used as a 12-plex DNA sensor array platform. DNA isolated from Salmonella was amplified by multiplex polymerase chain reaction (mPCR), and the presence of Salmonella was determined by reading fluorescent signals from hybridization between probes on DNA sensors and fluorescently labeled target DNA using the Bio-Plex® system. The developed multiplex array was able to detect synthetic DNA at the concentration as low as 100 fM and various Salmonella serovars as low as 100 CFU/mL within 1 h post-PCR. Sensitivity of this assay was further improved to 1 CFU/mL with 6-h enrichment. The array system also correctly and specifically identified serotype of tested Salmonella strains without any cross-reactivity with other common foodborne pathogens. Our results indicate the developed DNA sensor suspension array can be a rapid and reliable high-throughput method for simultaneous detection and molecular identification of common Salmonella serotypes.

  9. Gigapixel imaging with microlens arrays

    Science.gov (United States)

    Orth, Antony; Schonbrun, Ethan

    2016-03-01

    A crucial part of the drug discovery process involves imaging the response of thousands of cell cultures to candidate drugs. Quantitative parameters from these "high content screens", such as protein expression and cell morphology, are extracted from fluorescence and brightfield micrographs. Due to the sheer number of cells that need to imaged for adequate statistics, the imaging time itself is a major bottleneck. Automated microscopes image small fields-of-view (FOVs) serially, which are then stitched together to form gigapixel-scale mosaics. We have developed a microscopy architecture that reduces mechanical overhead of traditional large field-of-view by parallelizing the image capture process. Instead of a single objective lens imaging FOVs one by one, we employ a microlens array for continuous photon capture, resulting in a 3-fold throughput increase. In this contribution, we present the design and imaging results of this microscopy architecture in three different contrast modes: multichannel fluorescence, hyperspectral fluorescence and brightfield.

  10. A Portable Diode Array Spectrophotometer.

    Science.gov (United States)

    Stephenson, David

    2016-05-01

    A cheap portable visible light spectrometer is presented. The spectrometer uses readily sourced items and could be constructed by anyone with a knowledge of electronics. The spectrometer covers the wavelength range 450-725 nm with a resolution better than 5 nm. The spectrometer uses a diffraction grating to separate wavelengths, which are detected using a 128-element diode array, the output of which is analyzed using a microprocessor. The spectrum is displayed on a small liquid crystal display screen and can be saved to a micro SD card for later analysis. Battery life (2 × AAA) is estimated to be 200 hours. The overall dimensions of the unit are 120 × 65 × 60 mm, and it weighs about 200 g. © The Author(s) 2016.

  11. Array abstractions for GPU programming

    DEFF Research Database (Denmark)

    Dybdal, Martin

    The shift towards massively parallel hardware platforms for highperformance computing tasks has introduced a need for improved programming models that facilitate ease of reasoning for both users and compiler optimization. A promising direction is the field of functional data-parallel programming......, for which functional invariants can be utilized by optimizing compilers to perform large program transformations automatically. However, the previous work in this area allow users only limited ability to reason about the performance of algorithms. For this reason, such languages have yet to see wide...... industrial adoption. We present two programming languages that attempt at both supporting industrial applications and providing reasoning tools for hierarchical data-parallel architectures, such as GPUs. First, we present TAIL, an array based intermediate language and compiler framework for compiling a large...

  12. Conformable eddy current array delivery

    Science.gov (United States)

    Summan, Rahul; Pierce, Gareth; Macleod, Charles; Mineo, Carmelo; Riise, Jonathan; Morozov, Maxim; Dobie, Gordon; Bolton, Gary; Raude, Angélique; Dalpé, Colombe; Braumann, Johannes

    2016-02-01

    The external surface of stainless steel containers used for the interim storage of nuclear material may be subject to Atmospherically Induced Stress Corrosion Cracking (AISCC). The inspection of such containers poses a significant challenge due to the large quantities involved; therefore, automating the inspection process is of considerable interest. This paper reports upon a proof-of-concept project concerning the automated NDT of a set of test containers containing artificially generated AISCCs. An Eddy current array probe with a conformable padded surface from Eddyfi was used as the NDT sensor and end effector on a KUKA KR5 arc HW robot. A kinematically valid cylindrical raster scan path was designed using the KUKA|PRC path planning software. Custom software was then written to interface measurement acquisition from the Eddyfi hardware with the motion control of the robot. Preliminary results and analysis are presented from scanning two canisters.

  13. Scanning strategies for imaging arrays

    Science.gov (United States)

    Kovács, Attila

    2008-07-01

    Large-format (sub)millimeter wavelength imaging arrays are best operated in scanning observing modes rather than traditional position-switched (chopped) modes. The choice of observing mode is critical for isolating source signals from various types of noise interference, especially for ground-based instrumentation operating under a bright atmosphere. Ideal observing strategies can combat 1/f noise, resist instrumental defects, sensitively recover emission on large scales, and provide an even field coverage - all under feasible requirements of telescope movement. This work aims to guide the design of observing patterns that maximize scientific returns. It also compares some of the popular choices of observing modes for (sub)millimeter imaging, such as random, Lissajous, billiard, spiral, On-The-Fly (OTF), DREAM, chopped and stare patterns. Many of the conclusions are also applicable other imaging applications and imaging in one dimension (e.g. spectroscopic observations).

  14. LOFAR- The Low Frequency Array

    Science.gov (United States)

    Falcke, H. D. E.

    2006-08-01

    LOFAR is an innovative radio telescope in the frequency range of 10-240 MHz, realized as a phased array. It will become the largest radio telescope in the world in the time frame 2006-2010, located in Northern Europe. LOFAR is being implemented as a Wide Area Sensor Network which connects thousands of cheap sensors spread throughout the country to a central super computer using an ultra-broadband, synchronized data network. As the central processor IBM has provided its Blue Gene/L supercomputer. It will process streaming data with about 0.5 Terabit per second. Many simple radio antennas connected to the network turn it into a huge radio telescope for cosmological studies. In addition, geophones will turn LOFAR into an earthquake monitoring system and infrasound and meteorology sensors will turn LOFAR into a real-time weather monitoring array for agricultural applications. LOFAR is the first radio telescopes that can listen to radio signals from the entire sky overhead, on all time scales, at a large range of frequencies, and even look back in time for a couple of seconds. The main strength of LOFAR are surveys. One goal is to detect the first generation of black holes and galaxies in the universe during the epoch of reionization and study hydrogen formed after the big bang. LOFAR is also an ideal system to discover transient and sporadic radio signals. Likely transient sources to be discovered with LOFAR are bursting stars and Jupiter-like planets, gamma-ray bursts, radio outbursts from black holes, but also lightning on Earth and even radio flashes from ultra-high energy cosmic particles hitting the Earth atmosphere. Some LOFAR prototypes have recently been built. They have produced the first instantaneous all-sky maps and discovered the radio emission from cosmic particle air showers.

  15. Micromirror Array Control of a Phase-Locked Laser Diode Array

    Science.gov (United States)

    1995-12-01

    mirror. These mirrors were invented by Larry Hornbeck of Texas Instruments approximately 15 years ago. Two dimensional arrays of micromirrors are...AFIT/GAP/ENP/95D-2 MICROMIRROR ARRAY CONTROL OF A PHASE-LOCKED LASER DIODE ARRAY THESIS Carl J. Christensen, Captain, USAF AFIT/GAP/ENP/95D-2...Approved for public release; distribution unlimited. /99 (;/ Ig M 5 -5 DTiC QUALITY IITSYEGTED AFIT/GAP/ENP/95D-2 MICROMIRROR ARRAY CONTROL OF A PHASE-LOCKED

  16. Imaging spectroscopy using embedded diffractive optical arrays

    Science.gov (United States)

    Hinnrichs, Michele; Hinnrichs, Bradford

    2017-09-01

    Pacific Advanced Technology (PAT) has developed an infrared hyperspectral camera based on diffractive optic arrays. This approach to hyperspectral imaging has been demonstrated in all three infrared bands SWIR, MWIR and LWIR. The hyperspectral optical system has been integrated into the cold-shield of the sensor enabling the small size and weight of this infrared hyperspectral sensor. This new and innovative approach to an infrared hyperspectral imaging spectrometer uses micro-optics that are made up of an area array of diffractive optical elements where each element is tuned to image a different spectral region on a common focal plane array. The lenslet array is embedded in the cold-shield of the sensor and actuated with a miniature piezo-electric motor. This approach enables rapid infrared spectral imaging with multiple spectral images collected and processed simultaneously each frame of the camera. This paper will present our optical mechanical design approach which results in an infrared hyper-spectral imaging system that is small enough for a payload on a small satellite, mini-UAV, commercial quadcopter or man portable. Also, an application of how this spectral imaging technology can easily be used to quantify the mass and volume flow rates of hydrocarbon gases. The diffractive optical elements used in the lenslet array are blazed gratings where each lenslet is tuned for a different spectral bandpass. The lenslets are configured in an area array placed a few millimeters above the focal plane and embedded in the cold-shield to reduce the background signal normally associated with the optics. The detector array is divided into sub-images covered by each lenslet. We have developed various systems using a different number of lenslets in the area array. Depending on the size of the focal plane and the diameter of the lenslet array will determine the number of simultaneous different spectral images collected each frame of the camera. A 2 x 2 lenslet array will image

  17. Detection of Human Papillomavirus DNA in Cervical Samples: Analysis of the New PGMY-PCR Compared To the Hybrid Capture II and MY-PCR Assays and a Two-Step Nested PCR Assay

    OpenAIRE

    Giovannelli, Lucia; Lama, Anna; Capra, Giuseppina; Giordano, Viviana; Aricò, Pietro; Ammatuna, Pietro

    2004-01-01

    The PGMY-PCR for human papillomavirus (HPV) was evaluated, in parallel with nested PCR (nPCR), in samples with noted Hybrid Capture II (HCII) and MY-PCR results. PGMY-PCR detected HPV DNA in 2.5% of HCII-negative-MY-PCR-negative samples and in 71.7% of HCII-positive-MY-PCR-negative samples; also, it detected the MY-PCR-negative-nPCR-negative types HPV-42, HPV-44, HPV-51, HPV-87, and HPV-89.

  18. Digital PCR as a tool to measure HIV persistence.

    Science.gov (United States)

    Rutsaert, Sofie; Bosman, Kobus; Trypsteen, Wim; Nijhuis, Monique; Vandekerckhove, Linos

    2018-01-30

    Although antiretroviral therapy is able to suppress HIV replication in infected patients, the virus persists and rebounds when treatment is stopped. In order to find a cure that can eradicate the latent reservoir, one must be able to quantify the persisting virus. Traditionally, HIV persistence studies have used real-time PCR (qPCR) to measure the viral reservoir represented by HIV DNA and RNA. Most recently, digital PCR is gaining popularity as a novel approach to nucleic acid quantification as it allows for absolute target quantification. Various commercial digital PCR platforms are nowadays available that implement the principle of digital PCR, of which Bio-Rad's QX200 ddPCR is currently the most used platform in HIV research. Quantification of HIV by digital PCR is proving to be a valuable improvement over qPCR as it is argued to have a higher robustness to mismatches between the primers-probe set and heterogeneous HIV, and forfeits the need for a standard curve, both of which are known to complicate reliable quantification. However, currently available digital PCR platforms occasionally struggle with unexplained false-positive partitions, and reliable segregation between positive and negative droplets remains disputed. Future developments and advancements of the digital PCR technology are promising to aid in the accurate quantification and characterization of the persistent HIV reservoir.

  19. Dynamic array of dark optical traps

    DEFF Research Database (Denmark)

    Daria, V.R.; Rodrigo, P.J.; Glückstad, J.

    2004-01-01

    A dynamic array of dark optical traps is generated for simultaneous trapping and arbitrary manipulation of multiple low-index microstructures. The dynamic intensity patterns forming the dark optical trap arrays are generated using a nearly loss-less phase-to-intensity conversion of a phase-encode...

  20. Stochastic Beamforming via Compact Antenna Arrays

    DEFF Research Database (Denmark)

    Alrabadi, Osama; Pedersen, Gert Frølund

    2012-01-01

    The paper investigates the average beamforming (BF) gain of compact antenna arrays when statistical channel knowledge is available. The optimal excitation (precoding vector) and impedance termination that maximize the average BF gain are a compromise between the ones that maximize the array...

  1. Adaptive motion compensation in sonar array processing

    NARCIS (Netherlands)

    Groen, J.

    2006-01-01

    In recent years, sonar performance has mainly improved via a significant increase in array ap-erture, signal bandwidth and computational power. This thesis aims at improving sonar array processing techniques based on these three steps forward. In applications such as anti-submarine warfare and mine

  2. Element sharing in interleaved antenna arrays

    CSIR Research Space (South Africa)

    Du Plessis, WP

    2012-03-01

    Full Text Available The effect of allowing shared elements in interleaved thinned arrays is investigated. The sidelobe level (SLL) as a function of the number of shared elements mirrors the SLL of thinned arrays as a function of filling factor because the number...

  3. Refracting surface plasmon polaritons with nanoparticle arrays

    DEFF Research Database (Denmark)

    Radko, I.P.; Evlyukhin, A.B.; Boltasseva, Alexandra

    2008-01-01

    Refraction of surface plasmon polaritons (SPPs) by various structures formed by a 100-nm-period square lattice of gold nanoparticles on top of a gold film is studied by leakage radiation microscopy. SPP refraction by a triangular-shaped nanoparticle array indicates that the SPP effective refractive...... to design nanoparticle arrays for specific applications requiring in-plane SPP manipulation....

  4. Dumand-array data-acquisition system

    International Nuclear Information System (INIS)

    Brenner, A.E.; Theriot, D.; Dau, W.D.; Geelhood, B.D.; Harris, F.; Learned, J.G.; Stenger, V.; March, R.; Roos, C.; Shumard, E.

    1982-04-01

    An overall data acquisition approach for DUMAND is described. The scheme assumes one array to shore optical fiber transmission line for each string of the array. The basic event sampling period is approx. 13 μsec. All potentially interesting data is transmitted to shore where the major processing is performed

  5. The lofar phased array telescope system

    NARCIS (Netherlands)

    Gunst, André W.; Bentum, Marinus Jan

    2010-01-01

    The Low Frequency Array (LOFAR) is the largest telescope in the world operating at a frequency range from 30 to 240 MHz. LOFAR is the first radio telescope of its size which uses phased array principles to detect radio signals. More than 10,000 antennas are installed in the field. The antennas are

  6. Antennas for Frequency Reconfigurable Phased Arrays

    NARCIS (Netherlands)

    Haider, S.N.

    2015-01-01

    Sensors such as phased array radars play a crucial role in public safety. They are unavoidable for surveillance, threat identification and post-disaster management. However, different scenarios impose immensely diverse requirements for these systems. Phased array systems occupy a large space. In

  7. Overview arraying techniques for deep space communications

    Science.gov (United States)

    Mileant, A.; Hinedi, S.

    1994-01-01

    Four different arraying schemes applicable to deep space communications are discussed and analyzed. These include symbol stream combining (SSC), baseband combining (BC), carrier arraying (CA) and full spectrum combining (FSC). Complexity versus performance is traded off throughout the paper and benefits to the reception of existing spacecraft signals are discussed.

  8. Diagnostics of the BIOMASS feed array prototype

    DEFF Research Database (Denmark)

    Cappellin, Cecilia; Pivnenko, Sergey; Pontoppidan, Kennie Nybo

    2013-01-01

    The 3D reconstruction algorithm is applied to the prototype feed array of the BIOMASS synthetic aperture radar, recently measured at the DTU-ESA Spherical Near-Field Antenna Test Facility in Denmark. Careful analysis of the measured feed array data has shown that the test support structure...

  9. ALMA - the Atacama Large Millimeter Array

    NARCIS (Netherlands)

    Wild, W.; Cunningham, C.

    2002-01-01

    The Atacama Large Millimeter Array (ALMA) is a major ground based project for millimeter and submillimeter astronomy to be realized during this decade. It comprises an array of 64 telescopes of 12 meter diameter, each equipped with 10 receivers bands covering the atmospheric windows from 30 GHz to

  10. Spherical Horn Array for Wideband Propagation Measurements

    DEFF Research Database (Denmark)

    Franek, Ondrej; Pedersen, Gert Frølund

    2011-01-01

    properties and coupling between the elements are investigated via measurements and numerical simulations. Radiation patterns and sum beams of the array on selected frequencies throughout the band are also presented. Based on the obtained results it is concluded that the array is a good candidate...

  11. Maximum gain of Yagi-Uda arrays

    DEFF Research Database (Denmark)

    Bojsen, J.H.; Schjær-Jacobsen, Hans; Nilsson, E.

    1971-01-01

    Numerical optimisation techniques have been used to find the maximum gain of some specific parasitic arrays. The gain of an array of infinitely thin, equispaced dipoles loaded with arbitrary reactances has been optimised. The results show that standard travelling-wave design methods are not optimum...

  12. Law of substitution for mixed arrays

    International Nuclear Information System (INIS)

    Koudelka, A.J.

    1987-01-01

    The nuclear safety justification of a mixed array of dissimilar fissile units of metal units and dilute solution units, according to Clayton, has been a persistent and nagging problem. Dissimilar uranium metal or dissimilar uranium solution units in a mixed array can also create a modeling nightmare for the nuclear criticality safety engineer. Now, a calculational method known as the Law of Substitution has been developed to ensure that the k/sub eff/ of an array of uranium metal and uranium solution units will satisfy any k/sub eff/ limit set by the nuclear safety engineer. The nuclear criticality safety engineer can utilize the Law of Substitution to safely mix or substitute different uranium metal units, different uranium solution units, and more importantly, uranium metal and dilute UO 2 solution units in an array. The Law of Substitution is as follows: (1) calculate the k/sub eff/ of each unit type in its own infinite planar array. (2) Determine the edge-to-edge spacing of the infinite planar array of each type of unit to satisfy a desired k/sub eff/. (3) Select the largest edge-to-edge spacing from among the similar units in their infinite planar arrays and use that spacing for the finite or infinite planar array of mixed units

  13. The Astronomical Low-Frequency Array

    Science.gov (United States)

    Jones, D. L.; Allen, R. J.; Blume, W. H.; Desch, M. M.; Erickson, W. C.; Kaiser, M. L.; Kassim, N. E.; Kuiper, T. B. H.; Mahoney, M. J.; Marsh, K. A.; hide

    1996-01-01

    An array of satellites is proposed to make astronomic observations in the low frequency range of a few tens of MHz down to roughly 100 kHz, a range that cannot be observed through the ionosphere. The array would be in a solar orbit to avoid radio interference from Earth and to simplify trajectory tracking and control.

  14. Cloth-based hybridization array system for expanded identification of the animal species origin of derived materials in feeds.

    Science.gov (United States)

    Murphy, Johanna; Armour, Jennifer; Blais, Burton W

    2007-12-01

    A cloth-based hybridization array system (CHAS) previously developed for the detection of animal species for which prohibited materials have been specified (cattle, sheep, goat, elk, and deer) has been expanded to include the detection of animal species for which there are no prohibitions (pig and horse) in Canadian and American animal feeds. Animal species were identified by amplification of mitochondrial DNA sequences by PCR and subsequent hybridization of the amplicons with an array of species-specific oligonucleotide capture probes immobilized on a polyester cloth support, followed by an immunoenzymatic assay of the bound PCR products. The CHAS permitted sensitive and specific detection of meat meals from different animal species blended in a grain-based feed and should provide a useful adjunct to microscopic examination for the identification of prohibited materials in animal feeds.

  15. The Expanded Very Large Array Project

    Science.gov (United States)

    Perley, Rick

    2012-10-01

    The Expanded Very Large Array Project, begun in 2001, is now completed -- on time, on budget, and ``on spec.'' The major goal of the project was to multiply the key observational capabilities of the Very Large Array -- the world's most powerful, versatile, and productive radio telescope -- by a factor of at least an order of magnitude by implementation of modern technologies. The result of the project is an operationally new telescope -- renamed the Jansky Very Large Array -- which is capable of new science unimagined in 1980, when the original VLA was completed. In this talk, I will review the goals of the project, and describe the new capabilities of the Jansky Very Large Array. Already, the array is providing fabulous new insights into the physics of the universe,and I will spend the majority of the time describing examples of new results.

  16. Analyzing Array Manipulating Programs by Program Transformation

    Science.gov (United States)

    Cornish, J. Robert M.; Gange, Graeme; Navas, Jorge A.; Schachte, Peter; Sondergaard, Harald; Stuckey, Peter J.

    2014-01-01

    We explore a transformational approach to the problem of verifying simple array-manipulating programs. Traditionally, verification of such programs requires intricate analysis machinery to reason with universally quantified statements about symbolic array segments, such as "every data item stored in the segment A[i] to A[j] is equal to the corresponding item stored in the segment B[i] to B[j]." We define a simple abstract machine which allows for set-valued variables and we show how to translate programs with array operations to array-free code for this machine. For the purpose of program analysis, the translated program remains faithful to the semantics of array manipulation. Based on our implementation in LLVM, we evaluate the approach with respect to its ability to extract useful invariants and the cost in terms of code size.

  17. Milliarcsecond Astronomy with the CHARA Array

    Science.gov (United States)

    Schaefer, Gail; ten Brummelaar, Theo; Gies, Douglas; Jones, Jeremy; Farrington, Christopher

    2018-01-01

    The Center for High Angular Resolution Astronomy offers 50 nights per year of open access time at the CHARA Array. The Array consists of six telescopes linked together as an interferometer, providing sub-milliarcsecond resolution in the optical and near-infrared. The Array enables a variety of scientific studies, including measuring stellar angular diameters, imaging stellar shapes and surface features, mapping the orbits of close binary companions, and resolving circumstellar environments. The open access time is part of an NSF/MSIP funded program to open the CHARA Array to the broader astronomical community. As part of the program, we will build a searchable database for the CHARA data archive and run a series of one-day community workshops at different locations across the country to expand the user base for stellar interferometry and encourage new scientific investigations with the CHARA Array.

  18. COMPARISON OF 16S rRNA-PCR-RFLP, LipL32-PCR AND OmpL1-PCR METHODS IN THE DIAGNOSIS OF LEPTOSPIROSIS

    Directory of Open Access Journals (Sweden)

    Tülin GÜVEN GÖKMEN

    Full Text Available SUMMARY Leptospirosis is still one of the most important health problems in developing countries located in humid tropical and subtropical regions. Human infections are generally caused by exposure to water, soil or food contaminated with the urine of infected wild and domestic animals such as rodents and dogs. The clinical course of leptospirosis is variable and may be difficult to distinguish from many other infectious diseases. The dark-field microscopy (DFM, serology and nucleic acid amplification techniques are used to diagnose leptospirosis, however, a distinctive standard reference method is still lacking. Therefore, in this study, we aimed to determine the presence of Leptospira spp., to differentiate the pathogenic L. interrogans and the non-pathogenic L. biflexa, and also to determine the sensitivity and specificity values of molecular methods as an alternative to conventional ones. A total of 133 serum samples, from 47 humans and 86 cattle were evaluated by two conventional tests: the Microagglutination Test (MAT and the DFM, as well as three molecular methods, the 16S rRNA-PCR followed by Restriction Fragment Lenght Polymorphism (RFLP of the amplification products 16S rRNA-PCR-RFLP, LipL32-PCR and OmpL1-PCR. In this study, for L. interrogans, the specificity and sensitivity rates of the 16S rRNA-PCR and the LipL32-PCR were considered similar (100% versus 98.25% and 100% versus 98.68%, respectively. The OmpL1-PCR was able to classify L. interrogans into two intergroups, but this PCR was less sensitive (87.01% than the other two PCR methods. The 16S rRNA-PCR-RFLP could detect L. biflexa DNA, but LipL32-PCR and OmpL1-PCR could not. The 16S rRNA-PCR-RFLP provided an early and accurate diagnosis and was able to distinguish pathogenic and non-pathogenic Leptospira species, hence it may be used as an alternative method to the conventional gold standard techniques for the rapid disgnosis of leptospirosis.

  19. Multiplex enrichment quantitative PCR (ME-qPCR): a high-throughput, highly sensitive detection method for GMO identification.

    Science.gov (United States)

    Fu, Wei; Zhu, Pengyu; Wei, Shuang; Zhixin, Du; Wang, Chenguang; Wu, Xiyang; Li, Feiwu; Zhu, Shuifang

    2017-04-01

    Among all of the high-throughput detection methods, PCR-based methodologies are regarded as the most cost-efficient and feasible methodologies compared with the next-generation sequencing or ChIP-based methods. However, the PCR-based methods can only achieve multiplex detection up to 15-plex due to limitations imposed by the multiplex primer interactions. The detection throughput cannot meet the demands of high-throughput detection, such as SNP or gene expression analysis. Therefore, in our study, we have developed a new high-throughput PCR-based detection method, multiplex enrichment quantitative PCR (ME-qPCR), which is a combination of qPCR and nested PCR. The GMO content detection results in our study showed that ME-qPCR could achieve high-throughput detection up to 26-plex. Compared to the original qPCR, the Ct values of ME-qPCR were lower for the same group, which showed that ME-qPCR sensitivity is higher than the original qPCR. The absolute limit of detection for ME-qPCR could achieve levels as low as a single copy of the plant genome. Moreover, the specificity results showed that no cross-amplification occurred for irrelevant GMO events. After evaluation of all of the parameters, a practical evaluation was performed with different foods. The more stable amplification results, compared to qPCR, showed that ME-qPCR was suitable for GMO detection in foods. In conclusion, ME-qPCR achieved sensitive, high-throughput GMO detection in complex substrates, such as crops or food samples. In the future, ME-qPCR-based GMO content identification may positively impact SNP analysis or multiplex gene expression of food or agricultural samples. Graphical abstract For the first-step amplification, four primers (A, B, C, and D) have been added into the reaction volume. In this manner, four kinds of amplicons have been generated. All of these four amplicons could be regarded as the target of second-step PCR. For the second-step amplification, three parallels have been taken for

  20. Spatial normalization of array-CGH data

    Directory of Open Access Journals (Sweden)

    Brennetot Caroline

    2006-05-01

    Full Text Available Abstract Background Array-based comparative genomic hybridization (array-CGH is a recently developed technique for analyzing changes in DNA copy number. As in all microarray analyses, normalization is required to correct for experimental artifacts while preserving the true biological signal. We investigated various sources of systematic variation in array-CGH data and identified two distinct types of spatial effect of no biological relevance as the predominant experimental artifacts: continuous spatial gradients and local spatial bias. Local spatial bias affects a large proportion of arrays, and has not previously been considered in array-CGH experiments. Results We show that existing normalization techniques do not correct these spatial effects properly. We therefore developed an automatic method for the spatial normalization of array-CGH data. This method makes it possible to delineate and to eliminate and/or correct areas affected by spatial bias. It is based on the combination of a spatial segmentation algorithm called NEM (Neighborhood Expectation Maximization and spatial trend estimation. We defined quality criteria for array-CGH data, demonstrating significant improvements in data quality with our method for three data sets coming from two different platforms (198, 175 and 26 BAC-arrays. Conclusion We have designed an automatic algorithm for the spatial normalization of BAC CGH-array data, preventing the misinterpretation of experimental artifacts as biologically relevant outliers in the genomic profile. This algorithm is implemented in the R package MANOR (Micro-Array NORmalization, which is described at http://bioinfo.curie.fr/projects/manor and available from the Bioconductor site http://www.bioconductor.org. It can also be tested on the CAPweb bioinformatics platform at http://bioinfo.curie.fr/CAPweb.

  1. Comparison of droplet digital PCR and quantitative real-time PCR in mcrA-based methanogen community analysis

    Directory of Open Access Journals (Sweden)

    Tae Gwan Kim

    2014-12-01

    Full Text Available Two different quantitative PCR platforms, droplet digital PCR (dd-PCR and quantitative real-time PCR (qPCR, were compared in a mcrA-based methanogen community assay that quantifies ten methanogen sub-groups. Both technologies exhibited similar PCR efficiencies over at least four orders of magnitude and the same lower limits of detection (8 copies μL-DNA extract−1. The mcrA-based methanogen communities in three full-scale anaerobic digesters were examined using the two technologies. dd-PCR detected seven groups from the digesters, while qPCR did five groups, indicating that dd-PCR is more sensitive for DNA quantification. Linear regression showed quantitative agreements between both of the technologies (R2 = 0.59–0.98 in the five groups that were concurrently detected. Principal component analysis from the two datasets consistently indicated a substantial difference in the community composition among the digesters and revealed similar levels of differentiation among the communities. The combined results suggest that dd-PCR is more promising for examining methanogenic archaeal communities in biotechnological processes.

  2. How to Combine ChIP with qPCR.

    Science.gov (United States)

    Asp, Patrik

    2018-01-01

    Chromatin immunoprecipitation (ChIP) coupled with quantitative PCR (qPCR) has in the last 15 years become a basic mainstream tool in genomic research. Numerous commercially available ChIP kits, qPCR kits, and real-time PCR systems allow for quick and easy analysis of virtually anything chromatin-related as long as there is an available antibody. However, the highly accurate quantitative dimension added by using qPCR to analyze ChIP samples significantly raises the bar in terms of experimental accuracy, appropriate controls, data analysis, and data presentation. This chapter will address these potential pitfalls by providing protocols and procedures that address the difficulties inherent in ChIP-qPCR assays.

  3. Real-time quantitative PCR, pathogen detection and MIQE.

    Science.gov (United States)

    Johnson, Gemma; Nolan, Tania; Bustin, Stephen A

    2013-01-01

    Nucleic acids are the ultimate biomarker and real-time PCR (qPCR) is firmly established as the method of choice for nucleic acid detection. Together, they allow the accurate, sensitive and specific identification of pathogens, and the use of qPCR has become routine in diagnostic laboratories. The reliability of qPCR-based assays relies on a combination of optimal sample selection, assay design and validation as well as appropriate data analysis and the "Minimal Information for the Publication of real-time PCR" (MIQE) guidelines aim to improve both the reliability of assay design as well as the transparency of reporting, essential conditions if qPCR is to remain the benchmark technology for molecular diagnosis.

  4. Optimized MOL-PCR for Characterization of Microbial Pathogens.

    Science.gov (United States)

    Wuyts, Véronique; Roosens, Nancy H C; Bertrand, Sophie; Marchal, Kathleen; De Keersmaecker, Sigrid C J

    2016-01-06

    Characterization of microbial pathogens is necessary for surveillance, outbreak detection, and tracing of outbreak sources. This unit describes a multiplex oligonucleotide ligation-PCR (MOL-PCR) optimized for characterization of microbial pathogens. With MOL-PCR, different types of markers, like unique sequences, single-nucleotide polymorphisms (SNPs) and indels, can be simultaneously analyzed in one assay. This assay consists of a multiplex ligation for detection of the markers, a singleplex PCR for signal amplification, and hybridization to MagPlex-TAG beads for readout on a Luminex platform after fluorescent staining. The current protocol describes the MOL-PCR, as well as methods for DNA isolation, probe design, and data interpretation and it is based on an optimized MOL-PCR assay for subtyping of Salmonella Typhimurium. Copyright © 2016 John Wiley & Sons, Inc.

  5. Nasopharyngeal viral PCR in immunosuppressed patients and its association with virus detection in bronchoalveolar lavage by PCR.

    Science.gov (United States)

    Lachant, Daniel J; Croft, Daniel P; McGrane Minton, Heather; Prasad, Paritosh; Kottmann, Robert M

    2017-08-01

    Pulmonary infiltrates are common in immunosuppressed patients. Bronchoscopy with bronchoalveolar lavage (BAL) is often used to evaluate their aetiology. However, it may not always be easily performed. Thus, alternative diagnostic strategies may be needed. There is limited data on the correlation of nasopharyngeal (NP) respiratory viral panel (RVP)-PCR testing compared with BAL. We aimed to identify the predictive value of NP RVP-PCR samples compared with samples obtained from BAL in immunosuppressed patients with pulmonary infiltrates. We conducted an observational retrospective study of immunosuppressed adults who underwent bronchoscopy in the Pulmonary Department at the University of Rochester Medical Center between January 2011 and June 2016. We compared the positive and negative predictive values, sensitivity, specificity and false negative rate of NP RVP-PCR and BAL RVP-PCR, as well as identified clinical predictors of positive viral BAL RVP-PCR. Eighty-nine immunosuppressed patients had both NP and bronchoalveolar RVP-PCR testing. Twenty-one patients had NP(+)BAL(+) RVP-PCR testing. Seven patients had false negative (NP(-)BAL(+)) RVP-PCR testing. Three patients had NP(+)BAL(-) RVP-PCR testing. The positive and negative predictive values of NP RVP-PCR testing were 88% and 89%, respectively. Allogeneic bone marrow transplantation and testing performed in the winter and spring months were significantly associated with positive BAL RVP-PCR (OR = 3.3 (1.19-9.12); OR = 4.62 (1.64-12.99), respectively). NP RVP-PCR testing has high concordance with testing performed on BAL samples. Repeat testing through BAL is beneficial when there is high concern for viral infection after initial NP RVP-PCR testing is negative. © 2017 Asian Pacific Society of Respirology.

  6. Synthesis of unequally spaced linear antenna arrays | Hassen ...

    African Journals Online (AJOL)

    Synthesis of unequally spaced linear antenna arrays. MR Hassen, W Woldemariam ... Also the method needs fewer number of array elements to synthesize an array exhibiting the same (or better) level of directivity and sidelobe level in comparison with uniform arrays. Reduction in number of array elements has a positive ...

  7. 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.

  8. AGATA - Advanced GAmma Tracking Array

    Energy Technology Data Exchange (ETDEWEB)

    Akkoyun, S. [Department of Physics, Faculty of Science, Ankara University, 06100 Tandogan, Ankara (Turkey); Algora, A. [IFIC, CSIC-Universitat de Valencia, E-46980 Paterna (Spain); Alikhani, B. [IKP, TU Darmstadt, Schlossgartenstrasse 9, D-64289 Darmstadt (Germany); Ameil, F. [GSI Helmholtzzentrum fuer Schwerionenforschung GmbH, D-64291 Darmstadt (Germany); Angelis, G. de [INFN Laboratori Nazionali di Legnaro, IT-35020 Padova (Italy); Arnold, L. [Universite de Strasbourg, IPHC, 23 rue du Loess, 67037 Strasbourg (France); CNRS, UMR 7178, 67037 Strasbourg (France); Astier, A. [CSNSM, CNRS, IN2P3, Universite Paris-Sud, F-91405 Orsay (France); Atac, A. [Department of Physics, Faculty of Science, Ankara University, 06100 Tandogan, Ankara (Turkey); Department of Physics and Astronomy, Uppsala University, Uppsala (Sweden); Royal Institute of Technology, SE-10691 Stockholm (Sweden); Aubert, Y. [IPNO, CNRS/IN2P3, Universite Paris-Sud, F-91406 Orsay (France); Aufranc, C. [Universite de Lyon, Universite Lyon 1, CNRS-IN2P3, Institut de Physique Nucleaire de Lyon, F-69622 Villeurbanne (France); Austin, A. [STFC Daresbury Laboratory, Daresbury, Warrington WA4 4AD (United Kingdom); Aydin, S. [INFN Sezione di Padova, IT-35131 Padova (Italy); Azaiez, F. [IPNO, CNRS/IN2P3, Universite Paris-Sud, F-91406 Orsay (France); Badoer, S. [INFN Laboratori Nazionali di Legnaro, IT-35020 Padova (Italy); Balabanski, D.L. [Institute for Nuclear Research and Nuclear Energy, Bulgarian Academy of Sciences, Sofia (Bulgaria); Barrientos, D. [IFIC, CSIC-Universitat de Valencia, E-46980 Paterna (Spain); and others

    2012-03-11

    The Advanced GAmma Tracking Array (AGATA) is a European project to develop and operate the next generation {gamma}-ray spectrometer. AGATA is based on the technique of {gamma}-ray energy tracking in electrically segmented high-purity germanium crystals. This technique requires the accurate determination of the energy, time and position of every interaction as a {gamma} ray deposits its energy within the detector volume. Reconstruction of the full interaction path results in a detector with very high efficiency and excellent spectral response. The realisation of {gamma}-ray tracking and AGATA is a result of many technical advances. These include the development of encapsulated highly segmented germanium detectors assembled in a triple cluster detector cryostat, an electronics system with fast digital sampling and a data acquisition system to process the data at a high rate. The full characterisation of the crystals was measured and compared with detector-response simulations. This enabled pulse-shape analysis algorithms, to extract energy, time and position, to be employed. In addition, tracking algorithms for event reconstruction were developed. The first phase of AGATA is now complete and operational in its first physics campaign. In the future AGATA will be moved between laboratories in Europe and operated in a series of campaigns to take advantage of the different beams and facilities available to maximise its science output. The paper reviews all the achievements made in the AGATA project including all the necessary infrastructure to operate and support the spectrometer.

  9. Handheld ultrasound array imaging device

    Science.gov (United States)

    Hwang, Juin-Jet; Quistgaard, Jens

    1999-06-01

    A handheld ultrasound imaging device, one that weighs less than five pounds, has been developed for diagnosing trauma in the combat battlefield as well as a variety of commercial mobile diagnostic applications. This handheld device consists of four component ASICs, each is designed using the state of the art microelectronics technologies. These ASICs are integrated with a convex array transducer to allow high quality imaging of soft tissues and blood flow in real time. The device is designed to be battery driven or ac powered with built-in image storage and cineloop playback capability. Design methodologies of a handheld device are fundamentally different to those of a cart-based system. As system architecture, signal and image processing algorithm as well as image control circuit and software in this device is deigned suitably for large-scale integration, the image performance of this device is designed to be adequate to the intent applications. To elongate the battery life, low power design rules and power management circuits are incorporated in the design of each component ASIC. The performance of the prototype device is currently being evaluated for various applications such as a primary image screening tool, fetal imaging in Obstetrics, foreign object detection and wound assessment for emergency care, etc.

  10. Redundant arrays of IDE drives

    Energy Technology Data Exchange (ETDEWEB)

    D.A. Sanders et al.

    2002-01-02

    The authors report tests of redundant arrays of IDE disk drives for use in offline high energy physics data analysis. Parts costs of total systems using commodity EIDE disks are now at the $4000 per Terabyte level. Disk storage prices have now decreased to the point where they equal the cost per Terabyte of Storage Technology tape silos. The disks, however, offer far better granularity; even small institutions can afford to deploy systems. The tests include reports on software RAID-5 systems running under Linux 2.4 using Promise Ultra 100{trademark} disk controllers. RAID-5 protects data in case of a single disk failure by providing parity bits. Tape backup is not required. Journaling file systems are used to allow rapid recovery from crashes. The data analysis strategy is to encapsulate data and CPU processing power. Analysis for a particular part of a data set takes place on the PC where the data resides. The network is only used to put results together. They explore three methods of moving data between sites; internet transfers, not pluggable IDE disks in FireWire cases, and DVD-R disks.

  11. Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial

    DEFF Research Database (Denmark)

    Woudstra, C.; Skarin, H.; Anniballi, F.

    2013-01-01

    Two real-time PCR arrays based on the GeneDisc® cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated wi...

  12. Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial

    NARCIS (Netherlands)

    Woudstra, C.; Skarin, H.; Anniballi, F.; Auricchio, B.; Medici, De D.; Bano, L.; Drigo, I.; Hansen, T.; Löfström, Ch.; Hamidjaja, R.; Rotterdam, van B.; Koene, M.G.J.

    2013-01-01

    Two real-time PCR arrays based on the GeneDisc(®) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated

  13. Arrayed primer extension in the "array of arrays" format: a rational approach for microarray-based SNP genotyping

    DEFF Research Database (Denmark)

    Klitø, Niels G F; Tan, Qihua; Nyegaard, Mette

    2007-01-01

    This study provides a new version of the arrayed primer extension (APEX) protocol adapted to the 'array of arrays' platform using an instrumental setup for microarray processing not previously described. The primary aim of the study is to implement a system for rational cost-efficient genotyping...... where multiple singlenucleotide polymorphisms (SNPs) and individuals are genotyped on each microarray slide. Genotyping results are collected across 185 healthy Danish subjects and 76 SNPs on chromosome 3q13.31, because linkage to atopic disease phenotypes have been suggested in the Danish population...

  14. Highly precise measurement of HIV DNA by droplet digital PCR.

    Directory of Open Access Journals (Sweden)

    Matthew C Strain

    Full Text Available Deoxyribonucleic acid (DNA of the human immunodeficiency virus (HIV provides the most sensitive measurement of residual infection in patients on effective combination antiretroviral therapy (cART. Droplet digital PCR (ddPCR has recently been shown to provide highly accurate quantification of DNA copy number, but its application to quantification of HIV DNA, or other equally rare targets, has not been reported. This paper demonstrates and analyzes the application of ddPCR to measure the frequency of total HIV DNA (pol copies per million cells, and episomal 2-LTR (long terminal repeat circles in cells isolated from infected patients. Analysis of over 300 clinical samples, including over 150 clinical samples assayed in triplicate by ddPCR and by real-time PCR (qPCR, demonstrates a significant increase in precision, with an average 5-fold decrease in the coefficient of variation of pol copy numbers and a >20-fold accuracy improvement for 2-LTR circles. Additional benefits of the ddPCR assay over qPCR include absolute quantification without reliance on an external standard and relative insensitivity to mismatches in primer and probe sequences. These features make digital PCR an attractive alternative for measurement of HIV DNA in clinical specimens. The improved sensitivity and precision of measurement of these rare events should facilitate measurements to characterize the latent HIV reservoir and interventions to eradicate it.

  15. Detection and quantification of chimerism by droplet digital PCR.

    Science.gov (United States)

    George, David; Czech, Juliann; John, Bobby; Yu, Min; Jennings, Lawrence J

    2013-01-01

    Accurate quantification of chimerism and microchimerism is proving to be increasingly valuable for hematopoietic cell transplantation as well as non-transplant conditions. However, methods that are available to quantify low-level chimerism lack accuracy. Therefore, we developed and validated a method for quantifying chimerism based on digital PCR technology. We demonstrate accurate quantification that far exceeds what is possible with analog qPCR down to 0.01% with the potential to go even lower. Also, this method is inherently more informative than qPCR. We expect the advantages of digital PCR will make it the preferred method for chimerism analysis.

  16. Standardization of diagnostic PCR for the detection of foodborne pathogens

    DEFF Research Database (Denmark)

    Malorny, B.; Tassios, P.T.; Radstrom, P.

    2003-01-01

    neither gained wide acceptance in routine diagnostics nor been widely incorporated in standardized methods. Lack of validation and standard protocols, as well as variable quality of reagents and equipment, influence the efficient dissemination of PCR methodology from expert research laboratories to end......-user laboratories. Moreover, the food industry understandably requires and expects officially approved standards. Recognizing this, in 1999, the European Commission approved the research project, FOOD-PCR (http://www.PCR.dk), which aims to validate and standardize the use of diagnostic PCR for the detection...

  17. Application of hydroxylamine hydrochloride for post-PCR sterilization.

    Science.gov (United States)

    Aslanzadeh, J

    1993-04-01

    A major problem in the application of polymerase chain reaction (PCR) in diagnostic laboratories is contamination with exogenous nucleic acid, especially aerosolized amplicon, from previous PCR. Although several pre- and post-PCR sterilization techniques have been proposed, an optimal sterilization technique is not yet available. Hydroxylamine hydrochloride is a mutagenic agent that binds to and chemically modifies DNA. In the present study PCR was performed on DNA extracted from Herpes simplex virus (HSV) and Borrelia burgdorferi with two sets of primers that amplified a 92 bp sequence unique to HSV DNA polymerase gene and a 156 bp sequence unique to B. burgdorferi Osp-A gene (35 cycles). Following the amplification, PCR products were treated with 0-500 mM hydroxylamine hydrochloride and incubated at room temperature for 30 min. One microlitre of each hydroxylamine treated PCR product was reamplified for an additional 35 cycles. Pre- and post-hydroxylamine treated PCR products were separated by electrophoresis in 3% agarose gel. Hydroxylamine, at a concentration of 250 mM or higher, was found to effectively modify PCR products and prevent their amplification in subsequent PCR.

  18. Quantitative real-time PCR data analysis with R

    OpenAIRE

    Machado de Souza, Ignacio Nicolás

    2017-01-01

    This dissertation covers an introduction to the field of qPCR data analysis using the state-of-the-art R programming language. qPCR analyses genomic data based on the DNA replication. After showing a set of basic biological and statistics concepts, the roots of qPCR technology, together with its principal data analysis modelisations (visualisation, filtering, clustering, ...) are shown. A web application has been developed to ease and extend qPCR data analysis functionalities to other areas s...

  19. Assessment of Phthalate-induced Changes in Fetal Rat Testis Gene Expression using an rt-PCR Drug Metabolism Array.

    Science.gov (United States)

    Phthalate esters are a large family of compounds used in many industrial and commercial products. Based on numerous studies, phthalates such as diethyl hexyl phthalate (DEHP) produce reproductive malformations in male rodents through reduction of testosterone production and gene ...

  20. Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A

    OpenAIRE

    Ferreira, HL; Spilki, FR; dos Santos, MMAB; de Almeida, RS; Arns, CW

    2009-01-01

    Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an establis...

  1. Development of a peptide nucleic acid array platform for the detection of genetically modified organisms in food.

    Science.gov (United States)

    Germini, Andrea; Rossi, Stefano; Zanetti, Alessandro; Corradini, Roberto; Fogher, Corrado; Marchelli, Rosangela

    2005-05-18

    Two previously developed platforms, a multiplex polymerase chain reaction (PCR) and a peptide nucleic acid (PNA) array, the former allowing for the simultaneous detection of five transgenes and two endogenous controls in food and feed matrices and the latter for the assessment of the identity of amplified PCR products, were combined in order to develop a PNA array device for the screening of genetically modified organisms (GMOs) in food. PNA probes were opportunely designed, synthesized, and deposited on commercial slides. The length of the probes as well as the distance of the probes from the surface were evaluated and found to be critical points. The most suitable probes were found to be 15-mer PNAs linked to the slide surface by means of two 2-(2-aminoethoxy)ethoxyacetic acids as spacers. The device was tested on a model system constituted by flour samples containing a mixture of standards at known concentrations of transgenic material, in particular Roundup Ready soybean and Bt11, Bt176, Mon810, and GA21 maize: The DNA was amplified using the specific multiplex PCR method and tested on the PNA array. The method proposed was found to be able to correctly identify every GMO present in the tested samples.

  2. Geochemical Arrays at Woolsey Mound Seafloor Observatory

    Science.gov (United States)

    Sleeper, K.; Wilson, R. M.; Chanton, J.; Lapham, L.; Farr, N.; Camilli, R.; Martens, C. S.; Pontbriand, C.

    2011-12-01

    A suite of geochemical monitoring arrays has been developed for the Woolsey Mound Seafloor Observatory in the northern Gulf of Mexico to evaluate the oceanographic and tectonic forcing factors on the formation and stability of gas hydrates. These arrays are designed to collect sustained, time-series data of chemical concentrations, gradients and fluxes from the subsurface to the seafloor and into the near bottom water column. A Pore Fluid Array provides time-series measurements of methane, sulfate and salinity in subsurface pore waters to evaluate microbial activity, hydrate formation and/or hydrate dissociation. A Chimney Sampler Array collects in situ chemical and physical readings at the benthic boundary. The array is designed around a vertical cylinder with a known volume and washout rate for measuring chemical gradients and flux at the seafloor. The Benthic Boundary Layer Array extends into the water column with a package of sensors in a node close to the seafloor and a similar node 20 m above the seafloor to evaluate upward, downward and transversely advecting fluids. The three arrays can be used in concert to evaluate a release of methane by the dissociation of gas hydrates: the Pore Fluid Array identifies the breakdown of gas hydrates in the subsurface, the Chimney Array determines the rate of flux at the seafloor and the Benthic Boundary Layer Array evaluates the fate of the release in the water column. Combining the data from the geochemical arrays with output from the geophysical arrays provides key information to evaluate the specific and relative importance of tectonic and oceanographic triggers for hydrate dissociation. New probes and deployment platforms have been developed for the installation and maintenance of the arrays and new systems are in place and under development for the recovery of the data. Generally, the complete array or its components have to be recovered to download the data. However, this summer 2011, a new optic modem system was

  3. Tubular fluoropolymer arrays with high piezoelectric response

    Science.gov (United States)

    Zhukov, Sergey; Eder-Goy, Dagmar; Biethan, Corinna; Fedosov, Sergey; Xu, Bai-Xiang; von Seggern, Heinz

    2018-01-01

    Polymers with electrically charged internal air cavities called ferroelectrets exhibit a pronounced piezoelectric effect and are regarded as soft functional materials suitable for sensor and actuator applications. In this work, a simple method for fabricating piezoelectret arrays with open-tubular channels is introduced. A set of individual fluoroethylenepropylene (FEP) tubes is compressed between two heated metal plates. The squeezed FEP tubes are melted together at +270 °C. The resulting structure is a uniform, multi-tubular, flat array that reveals a strong piezoelectric response after a poling step. The fabricated arrays have a high ratio between piezoelectrically active and non-active areas. The optimal charging voltage and stability of the piezoelectric coefficients with pressures and frequency were experimentally investigated for two specific array structures with wall thickness of 50 and 120 μm. The array fabricated from 50 μm thick FEP tubes reveals a stable and high piezoelectric coefficient of {d}33 = 120-160 pC N-1 with a flat frequency response between 0.1 Hz and 10 kHz for pressures between 1 and 100 kPa. An increase of wall thickness to 120 μm is accompanied by a more than twofold decrease in the piezoelectric coefficient as a result of a simultaneously higher effective array stiffness and lower remanent polarization. The obtained experimental results can be used to optimize the array design with regard to the electromechanical performance.

  4. Comparison of nested PCR and real time PCR of Herpesvirus infections of central nervous system in HIV patients

    OpenAIRE

    Drago, Lorenzo; Lombardi, Alessandra; De Vecchi, Elena; Giuliani, Giuseppe; Bartolone, Rosaria; Gismondo, Maria Rita

    2004-01-01

    Abstract Background Molecular detection of herpesviruses DNA is considered as the reference standard assay for diagnosis of central nervous system infections. In this study nested PCR and real time PCR techniques for detection of Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in cerebrospinal fluid of HIV patients were compared. Methods Forty-six, 85 and 145 samples previously resulted positive for HSV-1, CMV and EBV by nested PCR and 150 randomly chos...

  5. Comparison of kDNA PCR-hybridization assay with three PCR methods for canines visceral Leishmaniasis diagnosis

    Energy Technology Data Exchange (ETDEWEB)

    Pilatti, Marcia M.; Andrade, Antero S.R. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: marciapilatti@yahoo.com.br, e-mail: antero@cdtn.br; Ferreira, Sidney A. [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Parasitologia], e-mail: saninoalmeida@gmail.com

    2009-07-01

    The sensitivity of the kDNA PCR-Hybridization assay, which uses radioactive DNA probes (labeled with {sup 32}P), was compared with three conventional PCR methods used for canine visceral leishmaniasis diagnosis. All PCR methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or semi nested). Two methods (kDNA PCR-Hybridization and kDNA snPCR) used primers addressed to kinetoplast minicircles and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of the ribosomal rRNA genes. The comparison was accomplished in two groups of 23 infected dogs using samples collected by the conjunctival swab procedure. In the Group 1 the DNA was extracted from cotton swabs by phenol-chloroform and in Group 2 by boiling. The most efficient PCR methods in the Group 1 were those based on kDNA targets. The kDNA PCR-Hybridization was able to detect parasites in 22/23 dogs (95.6%) and in 40/46 samples (86.9%). The kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The positivities of the kDNA based methods were significantly higher than the positivities verified for the methods based on ribosomal rRNA genes (p<0.05). In the Group 2 the kDNA PCR- Hybridization showed a better performance detecting parasites in 18/23 dogs (78.3%) and in 31/46 samples (67.4%), significantly higher than the other three methods (p<0.05). The higher sensitivity of the minicircle kDNA based assays reported by others was confirmed in this study and kDNA PCR-Hybridization showed the best sensitivity among the assays evaluated. (author)

  6. Comparison of kDNA PCR-hybridization assay with three PCR methods for canines visceral Leishmaniasis diagnosis

    International Nuclear Information System (INIS)

    Pilatti, Marcia M.; Andrade, Antero S.R.; Ferreira, Sidney A.

    2009-01-01

    The sensitivity of the kDNA PCR-Hybridization assay, which uses radioactive DNA probes (labeled with 32 P), was compared with three conventional PCR methods used for canine visceral leishmaniasis diagnosis. All PCR methods had two steps: a first amplification followed by hybridization or by a new amplification (nested or semi nested). Two methods (kDNA PCR-Hybridization and kDNA snPCR) used primers addressed to kinetoplast minicircles and the other two methods to the coding (LnPCR) and intergenic noncoding regions (ITS-1 nPCR) of the ribosomal rRNA genes. The comparison was accomplished in two groups of 23 infected dogs using samples collected by the conjunctival swab procedure. In the Group 1 the DNA was extracted from cotton swabs by phenol-chloroform and in Group 2 by boiling. The most efficient PCR methods in the Group 1 were those based on kDNA targets. The kDNA PCR-Hybridization was able to detect parasites in 22/23 dogs (95.6%) and in 40/46 samples (86.9%). The kDNA snPCR was positive for 21/23 dogs (91.3%) and for 40/46 samples (86.9%). The positivities of the kDNA based methods were significantly higher than the positivities verified for the methods based on ribosomal rRNA genes (p<0.05). In the Group 2 the kDNA PCR- Hybridization showed a better performance detecting parasites in 18/23 dogs (78.3%) and in 31/46 samples (67.4%), significantly higher than the other three methods (p<0.05). The higher sensitivity of the minicircle kDNA based assays reported by others was confirmed in this study and kDNA PCR-Hybridization showed the best sensitivity among the assays evaluated. (author)

  7. Detection of Campylobacter spp. in chilled and frozen broiler carcasses comparing immunoassay, PCR and real time PCR methods

    OpenAIRE

    Reis, Luciana Pimenta; Menezes, Liliane Denize Miranda; Lima, Graciela Kunrath; Santos, Ethiene Luiza de Souza; Dorneles, Elaine Maria Seles; Assis, Débora Cristina Sampaio de; Lage, Andrey Pereira; Cançado, Silvana de Vasconcelos; Figueiredo, Tadeu Chaves de

    2018-01-01

    ABSTRACT: In order to detect and identify Campylobacter spp. in broiler chicken carcasses, and to compare detection methods, 43 chilled and 43 frozen carcasses were collected and analyzed. Three methodologies were evaluated: an automated Enzyme Linked Fluorescent Assay (ELFA) VIDAS®30, Polymerase Chain Reaction (PCR) and real-time PCR. Only four chilled carcasses (4.6%) were considered positive for Campylobacter spp. by VIDAS®30 and no sample was positive when the conventional PCR technique w...

  8. A Microsphere-Based Suspension Array for Blood Group Molecular Typing: An Update

    Science.gov (United States)

    Drago, Francesca; Karpasitou, Katerina; Spinardi, Laura; Crespiatico, Loretta; Scalamogna, Mario; Poli, Francesca

    2010-01-01

    Summary Background In a previous publication we described a method for Jka/Jkb, Fya/Fyb, S/s, K/k, Kpa/Kpb, Jsa/Jsb, Coa/Cob, and Lua/Lub genotyping based on a microsphere suspension array. Here, an improved version of the assay is presented. Methods Two multiplex polymerase chain reactions (PCR) were developed: one for amplification of samples routinely tested and the other for those systems that are tested less frequently. Each biotinylated PCR product is hybridized in a single multiplex assay. A total of 2,020 samples were analyzed, and the genotypes were compared to the blood group phenotypes. Results There have been no discrepancies with the serology results other than null and/or weak phenotypes. Conclusion In its present form, the method presented here has the capacity to genotype hundreds of a samples in few hours with a high concordance rate with serology. PMID:21416028

  9. Large-scale nanophotonic phased array.

    Science.gov (United States)

    Sun, Jie; Timurdogan, Erman; Yaacobi, Ami; Hosseini, Ehsan Shah; Watts, Michael R

    2013-01-10

    Electromagnetic phased arrays at radio frequencies are well known and have enabled applications ranging from communications to radar, broadcasting and astronomy. The ability to generate arbitrary radiation patterns with large-scale phased arrays has long been pursued. Although it is extremely expensive and cumbersome to deploy large-scale radiofrequency phased arrays, optical phased arrays have a unique advantage in that the much shorter optical wavelength holds promise for large-scale integration. However, the short optical wavelength also imposes stringent requirements on fabrication. As a consequence, although optical phased arrays have been studied with various platforms and recently with chip-scale nanophotonics, all of the demonstrations so far are restricted to one-dimensional or small-scale two-dimensional arrays. Here we report the demonstration of a large-scale two-dimensional nanophotonic phased array (NPA), in which 64 × 64 (4,096) optical nanoantennas are densely integrated on a silicon chip within a footprint of 576 μm × 576 μm with all of the nanoantennas precisely balanced in power and aligned in phase to generate a designed, sophisticated radiation pattern in the far field. We also show that active phase tunability can be realized in the proposed NPA by demonstrating dynamic beam steering and shaping with an 8 × 8 array. This work demonstrates that a robust design, together with state-of-the-art complementary metal-oxide-semiconductor technology, allows large-scale NPAs to be implemented on compact and inexpensive nanophotonic chips. In turn, this enables arbitrary radiation pattern generation using NPAs and therefore extends the functionalities of phased arrays beyond conventional beam focusing and steering, opening up possibilities for large-scale deployment in applications such as communication, laser detection and ranging, three-dimensional holography and biomedical sciences, to name just a few.

  10. Study On Application Of Molecular Techniques (RAPD-PCR And RAMP-PCR) To Detect Mutation In Rice Breeding

    International Nuclear Information System (INIS)

    Hoang Thi My Linh; Phan, D. T. Son; Nguyen Thi Vang; Nguyen, T. T. Hien; Le XuanTham

    2007-01-01

    The project was carried out in 2007 with the purpose of consideration for using the two simple and inexpensive molecular techniques to estimate changes in DNA of rice mutant after gamma irradiation. Three rice cultivars: Basmati370, Tam Thom (TT1), IR64 and three gamma irradiated mutants BDS, TDS and VND 95-20 respectively, were used. Suitable DNA extraction procedure was obtained. PCR optimization was conducted on three important factors including: amount of MgCl 2 , DNA concentration and annealing temperature. 2.5 mM of MgCl 2 for RAPD-PCR and 3.75 mM for RAMP-PCR were found the best. 40 ng DNA provided a good amplification for RAMP-PCR; this figure was 50 ng for RAPD-PCR. Annealing temperatures were determined at 36 o C for RAPD primer and at 55±3 o C for Microsatellite primer. Final results showed that, both RAPD-PCR and RAMP-PCR could detect changes in DNA of rice mutants after gamma irradiation compared to their parents. Percentage of DNA changes determined by RAPD-PCR and RAMP-PCR on Basmati370 and its mutant BDS were 11.49% and 21.2% respectively; These on TT1 and TDS were 8.98% and 15.4%; and on IR64 and VND 95-20 were 3.45% and 4.95%. (author)

  11. Performance Analysis of Digital loudspeaker Arrays

    DEFF Research Database (Denmark)

    Pedersen, Bo Rohde; Kontomichos, Fotios; Mourjopoulos, John

    2008-01-01

    An analysis of digital loudspeaker arrays shows that the ways in which bits are mapped to the drivers influence the quality of the audio result. Specifically, a "bit-summed" rather than the traditional "bit-mapped" strategy greatly reduces the number of times drivers make binary transitions per...... period of the input frequency. Detailed simulations compare the results for a 32-loudspeaker array with a similar configuration with analog excitation of the drivers. Ideally, drivers in digital arrays should be very small and span a small area, but that sets limits on the low-frequency response...

  12. Arrays: The Heart and Soul of SIRTIF

    Science.gov (United States)

    Werner, Michael W.; Eisenhardt, Peter R.

    1993-01-01

    This paper describes the status of NASA's Space Infrared Telescope Facility (SIRTF) program. SIRTF will be a cryogenically cooled observatory for infrared astronomy from space and is planned for launch early in the next decade. It will be the first cryogenic observatory to make extensive use of the powerful infrared detector array technology discussed at this conference. We summarize a newly developed SIRTF mission concept and show how the availability of detector arrays has shaped the scientific rationale for SIRTF, and how the arrays themselves have become part of the definition of the SIRTF mission.

  13. Arrays: The heart and soul of SIRTF

    Science.gov (United States)

    Werner, Michael W.; Eisenhardt, Peter R.

    1994-01-01

    This pape describes the status of NASA's Space Infrared Telescope Facility (SIRTF) program. SIRTF will be a cryogenically cooled observatory for infrared astronomy from space and is planned for launch early in the next decade. It will be the first cryogenic space observatory to make extensive use of the powerful infrared detector array technology discussed at this conference. We summarize a newly developed SIRTF mission concept and show how the availability of detector arrays has shaped the scientific rationale for SIRTF, and how the arrays themselves have become part of the definition of the SIRTF mission.

  14. Flat-plate photovoltaic array design optimization

    Science.gov (United States)

    Ross, R. G., Jr.

    1980-01-01

    An analysis is presented which integrates the results of specific studies in the areas of photovoltaic structural design optimization, optimization of array series/parallel circuit design, thermal design optimization, and optimization of environmental protection features. The analysis is based on minimizing the total photovoltaic system life-cycle energy cost including repair and replacement of failed cells and modules. This approach is shown to be a useful technique for array optimization, particularly when time-dependent parameters such as array degradation and maintenance are involved.

  15. High-throughput simultaneous genotyping of human platelet antigen-1 to -16 by using suspension array.

    Science.gov (United States)

    An, Qun-Xing; Li, Cui-Ying; Xu, Li-Juan; Zhang, Xian-Qing; Bai, Yan-Jun; Shao, Zhong-Jun; Zhang, Wei

    2013-11-01

    Comprehensive and accurate detection of human platelet antigens (HPAs) plays a significant role in diagnosis and prevention of the platelet (PLT) alloimmune syndromes and ensuring clinical safety of patients undergoing PLT transfusion. The majority of the available methods are incapable of performing high-throughput simultaneous detection of HPA-1 to -16, and the accuracy of many methods needs to be further enhanced. We have developed a new HPA-genotyping method for simultaneous detection of HPA-1 to -16 based on suspension array technology. A total of 216 samples from Chinese Han donors in Xi'an were genotyped using the developed method, and all the samples again were genotyped using polymerase chain reaction (PCR) sequence-based typing (PCR-SBT), which is considered the gold standard. All 216 samples were successfully genotyped for HPA-1 to -16 using both our method and PCR-SBT. Results showed that the genotype and allele frequencies obtained using our method were fully consistent with those obtained using PCR-SBT. Our method provides accurate, high-throughput, and simultaneous genotyping of HPA-1 to -16 and will serve as the foundation for large-scale clinical genotyping of HPAs and for the establishment of an HPA-typed PLT donor registry. © 2013 American Association of Blood Banks.

  16. Improvements to previous algorithms to predict gene structure and isoform concentrations using Affymetrix Exon arrays

    Directory of Open Access Journals (Sweden)

    Aramburu Ander

    2010-11-01

    Full Text Available Abstract Background Exon arrays provide a way to measure the expression of different isoforms of genes in an organism. Most of the procedures to deal with these arrays are focused on gene expression or on exon expression. Although the only biological analytes that can be properly assigned a concentration are transcripts, there are very few algorithms that focus on them. The reason is that previously developed summarization methods do not work well if applied to transcripts. In addition, gene structure prediction, i.e., the correspondence between probes and novel isoforms, is a field which is still unexplored. Results We have modified and adapted a previous algorithm to take advantage of the special characteristics of the Affymetrix exon arrays. The structure and concentration of transcripts -some of them possibly unknown- in microarray experiments were predicted using this algorithm. Simulations showed that the suggested modifications improved both specificity (SP and sensitivity (ST of the predictions. The algorithm was also applied to different real datasets showing its effectiveness and the concordance with PCR validated results. Conclusions The proposed algorithm shows a substantial improvement in the performance over the previous version. This improvement is mainly due to the exploitation of the redundancy of the Affymetrix exon arrays. An R-Package of SPACE with the updated algorithms have been developed and is freely available.

  17. Fluid jet-array parallel machining of optical microstructure array surfaces.

    Science.gov (United States)

    Wang, Chunjin; Cheung, Chi Fai; Liu, Mingyu; Lee, Wing Bun

    2017-09-18

    Optical microstructure array surfaces such as micro-lens array surface, micro-groove array surface etc., are being used in more and more optical products, depending on its ability to produce a unique or particular performance. The geometrical complexity of the optical microstructures array surfaces makes them difficult to be fabricated. In this paper, a novel method named fluid jet-array parallel machining (FJAPM) is proposed to provide a new way to generate the microstructure array surfaces with high productivity. In this process, an array of abrasive water jets is pumped out of a nozzle, and each fluid jet simultaneously impinges the target surface to implement material removal independently. The jet-array nozzle was optimally designed firstly to diminish the effect of jet interference based on the experimental investigation on the 2-Jet nozzles with different jet intervals. The material removal and surface generation models were built and validated through the comparison of simulation and experimental results of the generation of several kinds of microstructure array surfaces. Following that, the effect of some factors in the process was discussed, including the fluid pressure, nozzle geometry, tool path, and dwell time. The experimental results and analysis prove that FJAPM process is an effective way to fabricate the optical microstructure array surface together with high productivity.

  18. A benchmark for microRNA quantification algorithms using the OpenArray platform.

    Science.gov (United States)

    McCall, Matthew N; Baras, Alexander S; Crits-Christoph, Alexander; Ingersoll, Roxann; McAlexander, Melissa A; Witwer, Kenneth W; Halushka, Marc K

    2016-03-22

    Several techniques have been tailored to the quantification of microRNA expression, including hybridization arrays, quantitative PCR (qPCR), and high-throughput sequencing. Each of these has certain strengths and limitations depending both on the technology itself and the algorithm used to convert raw data into expression estimates. Reliable quantification of microRNA expression is challenging in part due to the relatively low abundance and short length of the miRNAs. While substantial research has been devoted to the development of methods to quantify mRNA expression, relatively little effort has been spent on microRNA expression. In this work, we focus on the Life Technologies TaqMan OpenArray(Ⓡ) system, a qPCR-based platform to measure microRNA expression. Several algorithms currently exist to estimate expression from the raw amplification data produced by qPCR-based technologies. To assess and compare the performance of these methods, we performed a set of dilution/mixture experiments to create a benchmark data set. We also developed a suite of statistical assessments that evaluate many different aspects of performance: accuracy, precision, titration response, number of complete features, limit of detection, and data quality. The benchmark data and software are freely available via two R/Bioconductor packages, miRcomp and miRcompData. Finally, we demonstrate use of our software by comparing two widely used algorithms and providing assessments for four other algorithms. Benchmark data sets and software are crucial tools for the assessment and comparison of competing algorithms. We believe that the miRcomp and miRcompData packages will facilitate the development of new methodology for microRNA expression estimation.

  19. Protocols for 16S rDNA Array Analyses of Microbial Communities by Sequence-Specific Labeling of DNA Probes

    Directory of Open Access Journals (Sweden)

    Knut Rudi

    2003-01-01

    Full Text Available Analyses of complex microbial communities are becoming increasingly important. Bottlenecks in these analyses, however, are the tools to actually describe the biodiversity. Novel protocols for DNA array-based analyses of microbial communities are presented. In these protocols, the specificity obtained by sequence-specific labeling of DNA probes is combined with the possibility of detecting several different probes simultaneously by DNA array hybridization. The gene encoding 16S ribosomal RNA was chosen as the target in these analyses. This gene contains both universally conserved regions and regions with relatively high variability. The universally conserved regions are used for PCR amplification primers, while the variable regions are used for the specific probes. Protocols are presented for DNA purification, probe construction, probe labeling, and DNA array hybridizations.

  20. Verification of animal species in ham and salami by dna microarray and real time pcr methods

    Directory of Open Access Journals (Sweden)

    Zuzana Drdolová

    2017-01-01

    Full Text Available Consumer protection and detecting of adulteration is very important and has a wide societal impact in the economic sphere. Detection of animal species in meat products and the use of combining different methods is one of the means to achieve relevant product status. The aim of this study was to reveal whether or not the products label clearly meets the content declared by producer. In our study, 29 samples of meat products such as salami and ham obtained from stores and supermarkets in Slovakia were analyzed to detect the existing animal species according to the product label the use of Chipron LCD Array Analysis System, Meat 5.0. Products in which the presence of non-declared animal species has been detected were subjected to testing by the innuDETECT PCR Real-Time Kit, repeatedly. The results showed that 20 (68.96% samples were improperly labeled. From in total 14 tested ham samples 11 (78.57% products exhibited non-conformity with declared composition. Tested salami samples (15 revealed 9 (60% incorrectly labelled products. The results obtained by DNA Microarray and Real Time PCR methods were identical, and both methods should be extensively promoted for the detection of animal species in the meat and meat products. Normal 0 21 false false false EN-GB X-NONE X-NONE

  1. Multiplex RT-PCR and Automated Microarray for Detection of Eight Bovine Viruses.

    Science.gov (United States)

    Lung, O; Furukawa-Stoffer, T; Burton Hughes, K; Pasick, J; King, D P; Hodko, D

    2017-12-01

    Microarrays can be a useful tool for pathogen detection as it allow for simultaneous interrogation of the presence of a large number of genetic sequences in a sample. However, conventional microarrays require extensive manual handling and multiple pieces of equipment for printing probes, hybridization, washing and signal detection. In this study, a reverse transcription (RT)-PCR with an accompanying novel automated microarray for simultaneous detection of eight viruses that affect cattle [vesicular stomatitis virus (VSV), bovine viral diarrhoea virus type 1 and type 2, bovine herpesvirus 1, bluetongue virus, malignant catarrhal fever virus, rinderpest virus (RPV) and parapox viruses] is described. The assay accurately identified a panel of 37 strains of the target viruses and identified a mixed infection. No non-specific reactions were observed with a panel of 23 non-target viruses associated with livestock. Vesicular stomatitis virus was detected as early as 2 days post-inoculation in oral swabs from experimentally infected animals. The limit of detection of the microarray assay was as low as 1 TCID 50 /ml for RPV. The novel microarray platform automates the entire post-PCR steps of the assay and integrates electrophoretic-driven capture probe printing in a single user-friendly instrument that allows array layout and assay configuration to be user-customized on-site. © 2016 Her Majesty the Queen in Right of Canada.

  2. Inhibitory effect of common microfluidic materials on PCR outcome

    KAUST Repository

    Kodzius, Rimantas

    2013-10-10

    In this study, we established a simple method for evaluating the PCR compatibility of various common materials employed when fabricating microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most cases, adding bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, although they noticeably interacted with the polymerase. We provide a simple method of performing PCR-compatibility testing of materials using inexpensive instrumentation that is common in molecular biology laboratories. Furthermore, our method is direct, being performed under actual PCR conditions with high temperature. Our results provide an overview of materials that are PCR-friendly for fabricating microfluidic devices. The PCR reaction, without any additives, performed best with pyrex glass, and it performed worst with PMMA or acrylic glue materials.

  3. Multiplex polymerase chain reaction (PCR) on a SU-8 chip

    DEFF Research Database (Denmark)

    Christensen, Troels Balmer; Bang, Dang Duong; Wolff, Anders

    2008-01-01

    the PCR. The chip performs very well with respect to heating and cooling rates with values up to around 40 °C/s and 20 °C/s, respectively, and has low power consumption (0.5–2.5 W depending on temperature). Multiplex DNA amplification by PCR for the detection of Campylobacter at species level...

  4. (DNA) fragment using two-step polymerase chain reaction (PCR)

    African Journals Online (AJOL)

    Polymerase chain reaction (PCR), an essential tool in many fields such as molecular biology, normally comprises three steps: denaturation at a high temperature, annealing at a low temperature and elongation at a moderate temperature. Here, we report a two-step PCR method which incorporates annealing and elongation ...

  5. Clinical consequences of PCR based diagnosis of intestinal parasitic infections

    NARCIS (Netherlands)

    Rijsman, Lucas H; Monkelbaan, Jan F|info:eu-repo/dai/nl/344499383; Kusters, Johannes G|info:eu-repo/dai/nl/074307428

    2016-01-01

    The implementation of Polymerase Chain Reaction (PCR) based diagnostics of intestinal protozoa have led to higher sensitivity and (subtype) specificity, more convenient sampling and the possibility for high-throughput screening. An increasing number of clinical laboratories use PCR for routine

  6. Comparison between ICT and PCR for diagnosis of Chlamydia trachomatis.

    Science.gov (United States)

    Khan, E R; Hossain, M A; Paul, S K; Mahmud, C; Hasan, M M; Rahman, M M; Nahar, K; Kubayashi, N

    2012-04-01

    Chlamydia trachomatis is an obligate intracellular gram-negative bacterium which is the most prevalent cause of bacterial sexually transmitted infections (STI). The present study was carried to diagnose genital Chlamydia trachomatis infection among women of reproductive age, attending Mymensingh Medical College Hospital, during July 2009 to June 2010 by Immunochromatographic test (ICT) and Polymerase chain reaction (PCR). A total of 70 females were included in this study. Out of 70 cases 56 were symptomatic and 14 asymptomatic. Endocervical swabs were collected from each of the cases and examined by Immunochromatographic test (ICT) for antigen detection and Polymerase chain reaction (PCR) for detection of endogenous plasmid-based nucleic acid. A total 29(41.4%) of the cases were found positive for C. trachomatis either by ICT or PCR. Of the 56 symptomatic cases, 19(33.9%) were found ICT positive and 17(30.4%) were PCR positive. Among 14 asymptomatic females, 2(14.3%) were ICT positive and none were PCR positive. Though PCR is highly sensitive but a total of twelve cases were found ICT positive but PCR negative. It may be due to presence of plasmid deficient strain of C trachomatis which could be amplified by ompA based (Chromosomal gene) multiplex PCR.

  7. Evaluation of PCR for diagnosis of visceral leishmaniasis

    NARCIS (Netherlands)

    Osman, O. F.; Oskam, L.; Zijlstra, E. E.; Kroon, N. C.; Schoone, G. J.; Khalil, E. T.; El-Hassan, A. M.; Kager, P. A.

    1997-01-01

    An evaluation of Leishmania PCR was performed with bone marrow, lymph node, and blood samples from 492 patients, 60 positive controls, and 90 negative controls. Results were compared with microscopy results for Giemsa-stained smears. PCR and microscopy of lymph node and bone marrow aspirates from

  8. Real time PCR mediated determination of the spontaneous ...

    African Journals Online (AJOL)

    The study evaluates the utility of Real Time PCR (RT-PCR) in quantitative and qualitative analysis of alleles in sorghum populations and the spontaneous occurrence of Sorghum bicolor alleles in wild populations of sorghum. Leaf and seed material from wild sorghum accesions were sampled in Homabay, Siaya and Busia ...

  9. Species specific polymerase chain reaction (PCR) assay for ...

    African Journals Online (AJOL)

    A highly specific single step polymerase chain reaction (PCR) is described for the detection of pig (Sus domesticus) meat. A PCR assay was successfully optimized for amplification of 629 and 322-bp DNA fragment extracted from pig meat using designed species-specific primer pairs based on mitochondrial D-loop and 12S ...

  10. PCR-based detection of allergenic mackerel ingredients in seafood

    Indian Academy of Sciences (India)

    Administrator

    Keywords. Scomber mackerel; allergy; seafood; PCR; genus identification. RESEARCH NOTE. PCR-based detection of allergenic mackerel ingredients in seafood. FUTOSHI ARANISHI 1,* and TAKANE OKIMOTO 1,2. 1Department of Biological and Environmental Sciences, Miyazaki University,. Miyazaki 889-2192, Japan.

  11. Human fecal source identification with real-time quantitative PCR

    Science.gov (United States)

    Waterborne diseases represent a significant public health risk worldwide, and can originate from contact with water contaminated with human fecal material. We describe a real-time quantitative PCR (qPCR) method that targets a Bacteroides dori human-associated genetic marker for...

  12. Development of a competitive PCR assay for the quantification of ...

    African Journals Online (AJOL)

    The E. coli malate dehydrogenase Mdh house-keeping gene was modified and used as an internal control and competitor DNA for the c-PCR. E. coli cell concentration equivalents ranging from 20 to 2 x 104 cells ml-1 could be quantified with the c-PCR. Fifty-three water samples from various sources were tested with the ...

  13. PCR identification of Fusarium genus based on nuclear ribosomal ...

    African Journals Online (AJOL)

    PCR sensitivity ranged from 100 fg to 10 ng for DNA extracted from Fusarium oxysporum mycelium. No amplification products were detected with PCR of DNA from Rhizoctonia solani and Macrophomina phaseolina isolates using these primers. The assay is useful for rapid identification of Fusarium spp. cultures.

  14. Detection of Bacillus spores using PCR and FTA filters.

    Science.gov (United States)

    Lampel, Keith A; Dyer, Deanne; Kornegay, Leroy; Orlandi, Palmer A

    2004-05-01

    Emphasis has been placed on developing and implementing rapid detection systems for microbial pathogens. We have explored the utility of expanding FTA filter technology for the preparation of template DNA for PCR from bacterial spores. Isolated spores from several Bacillus spp., B. subtilis, B. cereus, and B. megaterium, were applied to FTA filters, and specific DNA products were amplified by PCR. Spore preparations were examined microscopically to ensure that the presence of vegetative cells, if any, did not yield misleading results. PCR primers SRM86 and SRM87 targeted a conserved region of bacterial rRNA genes, whereas primers Bsub5F and Bsub3R amplified a product from a conserved sequence of the B. subtilis rRNA gene. With the use of the latter set of primers for nested PCR, the sensitivity of the PCR-based assay was increased. Overall, 53 spores could be detected after the first round of PCR, and the sensitivity was increased to five spores by nested PCR. FTA filters are an excellent platform to remove PCR inhibitors and have universal applications for environmental, clinical, and food samples.

  15. Instrumentation and Fluorescent Chemistries Used in qPCR

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Löfström, Charlotta; Hansen, Trine

    2012-01-01

    -time PCR instrumentation, including the thermal and optical systems and the software. Performance parameters such as temperature uniformity, accuracy and ramp speed as well as reaction format, optical systems, calibration of dyes, software and comparison between different real-time PCR platforms...

  16. Detection of Tritrichomonas Foetus in Cattle Using a PCR ...

    African Journals Online (AJOL)

    A trichomoniasis polymerase chain reaction (PCR) assay was successfully established at the Central Veterinary Laboratory (CVL). Four preputial and 27 vaginal washings of bulls and cows obtained from communal cattle in Zimbabwe's Buhera and Wedza districts were tested using a PCR amplification assay as well as the ...

  17. Digital PCR for direct quantification of viruses without DNA extraction.

    Science.gov (United States)

    Pavšič, Jernej; Žel, Jana; Milavec, Mojca

    2016-01-01

    DNA extraction before amplification is considered an essential step for quantification of viral DNA using real-time PCR (qPCR). However, this can directly affect the final measurements due to variable DNA yields and removal of inhibitors, which leads to increased inter-laboratory variability of qPCR measurements and reduced agreement on viral loads. Digital PCR (dPCR) might be an advantageous methodology for the measurement of virus concentrations, as it does not depend on any calibration material and it has higher tolerance to inhibitors. DNA quantification without an extraction step (i.e. direct quantification) was performed here using dPCR and two different human cytomegalovirus whole-virus materials. Two dPCR platforms were used for this direct quantification of the viral DNA, and these were compared with quantification of the extracted viral DNA in terms of yield and variability. Direct quantification of both whole-virus materials present in simple matrices like cell lysate or Tris-HCl buffer provided repeatable measurements of virus concentrations that were probably in closer agreement with the actual viral load than when estimated through quantification of the extracted DNA. Direct dPCR quantification of other viruses, reference materials and clinically relevant matrices is now needed to show the full versatility of this very promising and cost-efficient development in virus quantification.

  18. Splinkerette PCR for mapping transposable elements in Drosophila.

    Directory of Open Access Journals (Sweden)

    Christopher J Potter

    2010-04-01

    Full Text Available Transposable elements (such as the P-element and piggyBac have been used to introduce thousands of transgenic constructs into the Drosophila genome. These transgenic constructs serve many roles, from assaying gene/cell function, to controlling chromosome arm rearrangement. Knowing the precise genomic insertion site for the transposable element is often desired. This enables identification of genomic enhancer regions trapped by an enhancer trap, identification of the gene mutated by a transposon insertion, or simplifying recombination experiments. The most commonly used transgene mapping method is inverse PCR (iPCR. Although usually effective, limitations with iPCR hinder its ability to isolate flanking genomic DNA in complex genomic loci, such as those that contain natural transposons. Here we report the adaptation of the splinkerette PCR (spPCR method for the isolation of flanking genomic DNA of any P-element or piggyBac. We report a simple and detailed protocol for spPCR. We use spPCR to 1 map a GAL4 enhancer trap located inside a natural transposon, pinpointing a master regulatory region for olfactory neuron expression in the brain; and 2 map all commonly used centromeric FRT insertion sites. The ease, efficiency, and efficacy of spPCR could make it a favored choice for the mapping of transposable element in Drosophila.

  19. Rapid detection of methicillin-resistant staphylococci by multiplex PCR

    African Journals Online (AJOL)

    A rapid and sensitive method for excluding the presence of methicillin-resistant Staphylococcus aureus (MRSA) in clinical samples was developed. The combination of MRSA detection by mecA coaA PCR with prior enrichment in selective broth was tested for 300 swabs. PCR identified 26 MRSApositive samples, ...

  20. Rapid detection of methicillin-resistant staphylococci by multiplex PCR

    African Journals Online (AJOL)

    Administrator

    2010-11-08

    Nov 8, 2010 ... A rapid and sensitive method for excluding the presence of methicillin-resistant Staphylococcus aureus (MRSA) in clinical samples was developed. The combination of MRSA detection by mecA coaA. PCR with prior enrichment in selective broth was tested for 300 swabs. PCR identified 26 MRSA- positive ...

  1. Polymerase chain reaction (PCR) detection of mixed trypanosome ...

    African Journals Online (AJOL)

    user

    2012-10-02

    Oct 2, 2012 ... kinase (KIN) and serum resistance- associated (SRA) primers, respectively for Trypanozoon group species, universal trypanosome ... Furthermore, PCR using serum resistance-associated. (SRA) gene primers which ..... PCR- based detection and typing of parasites. pp. 261-287. In: Parasitology for the 21st ...

  2. Conformal array design on arbitrary polygon surface with transformation optics

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Li, E-mail: dengl@bupt.edu.cn; Hong, Weijun, E-mail: hongwj@bupt.edu.cn; Zhu, Jianfeng; Peng, Biao; Li, Shufang [Beijing Key Laboratory of Network System Architecture and Convergence, School of Information and Communication Engineering, Beijing University of Posts and Telecommunications, 100876 Beijing (China); Wu, Yongle, E-mail: wuyongle138@gmail.com [Beijing Key Laboratory of Work Safety Intelligent Monitoring, School of Electronic Engineering, Beijing University of Posts and Telecommunications, 100876 Beijing (China)

    2016-06-15

    A transformation-optics based method to design a conformal antenna array on an arbitrary polygon surface is proposed and demonstrated in this paper. This conformal antenna array can be adjusted to behave equivalently as a uniformly spaced linear array by applying an appropriate transformation medium. An typical example of general arbitrary polygon conformal arrays, not limited to circular array, is presented, verifying the proposed approach. In summary, the novel arbitrary polygon surface conformal array can be utilized in array synthesis and beam-forming, maintaining all benefits of linear array.

  3. Luminometric Label Array for Counting and Differentiation of Bacteria.

    Science.gov (United States)

    Högmander, Milla; Paul, Catherine J; Chan, Sandy; Hokkanen, Elina; Eskonen, Ville; Pahikkala, Tapio; Pihlasalo, Sari

    2017-03-07

    Methods for simple and fast detection and differentiation of bacterial species are required, for instance, in medicine, water quality monitoring, and the food industry. Here, we have developed a novel label array method for the counting and differentiation of bacterial species. This method is based on the nonspecific interactions of multiple unstable lanthanide chelates and selected chemicals within the sample leading to a luminescence signal profile that is unique to the bacterial species. It is simple, cost-effective, and/or user-friendly compared to many existing methods, such as plate counts on selective media, automatic (hemocytometer-based) cell counters, flow cytometry, and polymerase chain reaction (PCR)-based methods for identification. The performance of the method was demonstrated with nine single strains of bacteria in pure culture. The limit of detection for counting was below 1000 bacteria per mL, with an average coefficient of variation of 10% achieved with the developed label array. A predictive model was trained with the measured luminescence signals and its ability to differentiate all tested bacterial species from each other, including members of the same genus Bacillus licheniformis and Bacillus subtilis, was confirmed via leave-one-out cross-validation. The suitability of the method for analysis of mixtures of bacterial species was shown with ternary mixtures of Bacillus licheniformis, Escherichia coli JM109, and Lactobacillus reuteri ATCC PTA 4659. The potential future application of the method could be monitoring for contamination in pure cultures; analysis of mixed bacterial cultures, where examining one species in the presence of another could inform industrial microbial processes; and the analysis of bacterial biofilms, where nonspecific methods based on physical and chemical characteristics are required instead of methods specific to individual bacterial species.

  4. 3C-digital PCR for quantification of chromatin interactions.

    Science.gov (United States)

    Du, Meijun; Wang, Liang

    2016-12-06

    Chromosome conformation capture (3C) is a powerful and widely used technique for detecting the physical interactions between chromatin regions in vivo. The principle of 3C is to convert physical chromatin interactions into specific DNA ligation products, which are then detected by quantitative polymerase chain reaction (qPCR). However, 3C-qPCR assays are often complicated by the necessity of normalization controls to correct for amplification biases. In addition, qPCR is often limited to a certain cycle number, making it difficult to detect fragment ligations with low frequency. Recently, digital PCR (dPCR) technology has become available, which allows for highly sensitive nucleic acid quantification. Main advantage of dPCR is its high precision of absolute nucleic acid quantification without requirement of normalization controls. To demonstrate the utility of dPCR in quantifying chromatin interactions, we examined two prostate cancer risk loci at 8q24 and 2p11.2 for their interaction target genes MYC and CAPG in LNCaP cell line. We designed anchor and testing primers at known regulatory element fragments and target gene regions, respectively. dPCR results showed that interaction frequency between the regulatory element and MYC gene promoter was 0.7 (95% CI 0.40-1.10) copies per 1000 genome copies while other regions showed relatively low ligation frequencies. The dPCR results also showed that the ligation frequencies between the regulatory element and two EcoRI fragments containing CAPG gene promoter were 1.9 copies (95% CI 1.41-2.47) and 1.3 copies per 1000 genome copies (95% CI 0.76-1.92), respectively, while the interaction signals were reduced on either side of the promoter region of CAPG gene. Additionally, we observed comparable results from 3C-dPCR and 3C-qPCR at 2p11.2 in another cell line (DU145). Compared to traditional 3C-qPCR, our results show that 3C-dPCR is much simpler and more sensitive to detect weak chromatin interactions. It may eliminate

  5. ISPA (imaging silicon pixel array) experiment

    CERN Multimedia

    Patrice Loïez

    2002-01-01

    Application components of ISPA tubes are shown: the CERN-developed anode chip, special windows for gamma and x-ray detection, scintillating crystal and fibre arrays for imaging and tracking of ionizing particles.

  6. Fiber Optic Pressure Sensor Array, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — VIP Sensors proposes to develop a Fiber Optic Pressure Sensor Array for measuring air flow pressure at multiple points on the skin of aircrafts for Flight Load Test...

  7. Marine Attack on Towed Hydrophone Arrays

    National Research Council Canada - National Science Library

    Kalmijn, Ad

    2002-01-01

    The original objective of the SIO Marine Attack project was to identify the electric and magnetic fields causing sharks to inflict serious damage upon the towed hydrophone arrays of US Navy submarines...

  8. Spacecraft Multiple Array Communication System Performance Analysis

    Science.gov (United States)

    Hwu, Shian U.; Desilva, Kanishka; Sham, Catherine C.

    2010-01-01

    The Communication Systems Simulation Laboratory (CSSL) at the NASA Johnson Space Center is tasked to perform spacecraft and ground network communication system simulations, design validation, and performance verification. The CSSL has developed simulation tools that model spacecraft communication systems and the space and ground environment in which the tools operate. In this paper, a spacecraft communication system with multiple arrays is simulated. Multiple array combined technique is used to increase the radio frequency coverage and data rate performance. The technique is to achieve phase coherence among the phased arrays to combine the signals at the targeting receiver constructively. There are many technical challenges in spacecraft integration with a high transmit power communication system. The array combining technique can improve the communication system data rate and coverage performances without increasing the system transmit power requirements. Example simulation results indicate significant performance improvement can be achieved with phase coherence implementation.

  9. Mid-Frequency Source Array Engineering Salaries

    National Research Council Canada - National Science Library

    Hodgkiss, William

    2003-01-01

    ... (time reversal "mirror") concepts. The source-receive array and its companion surface buoy operate in an autonomous fashion thus facilitating single ship operations for carrying out experimental work...

  10. Self-Deploying Tent Array, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — The Self-Deploying Tent Array (SDTA) is a modular power system that can be scaled to very large power levels for use on the Martian surface. The tent shape is...

  11. Strain Actuated Solar Arrays (SASA), Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — The team of CU Aerospace and the University of Illinois at Urbana-Champaign propose multifunctional solar arrays, which can be used for attitude control of a...

  12. Digital Array Gas Radiometer (DAGR) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The innovation proposed here is a digital array gas radiometer (DAGR), a new design for a gas filter correlation radiometer (GFCR) to accurately measure and monitor...

  13. Radiation Hardened Bolometer Linear Array Project

    Data.gov (United States)

    National Aeronautics and Space Administration — NASA has developed space-based thermal instrument spectrometers based on thermopile detectors linear arrays that are intrinsically radiation hard. Micro-bolometers...

  14. Statistical monitoring of linear antenna arrays

    KAUST Repository

    Harrou, Fouzi

    2016-11-03

    The paper concerns the problem of monitoring linear antenna arrays using the generalized likelihood ratio (GLR) test. When an abnormal event (fault) affects an array of antenna elements, the radiation pattern changes and significant deviation from the desired design performance specifications can resulted. In this paper, the detection of faults is addressed from a statistical point of view as a fault detection problem. Specifically, a statistical method rested on the GLR principle is used to detect potential faults in linear arrays. To assess the strength of the GLR-based monitoring scheme, three case studies involving different types of faults were performed. Simulation results clearly shown the effectiveness of the GLR-based fault-detection method to monitor the performance of linear antenna arrays.

  15. Study and Design of Differential Microphone Arrays

    CERN Document Server

    Benesty, Jacob

    2013-01-01

    Microphone arrays have attracted a lot of interest over the last few decades since they have the potential to solve many important problems such as noise reduction/speech enhancement, source separation, dereverberation, spatial sound recording, and source localization/tracking, to name a few. However, the design and implementation of microphone arrays with beamforming algorithms is not a trivial task when it comes to processing broadband signals such as speech. Indeed, in most sensor arrangements, the beamformer tends to have a frequency-dependent response. One exception, perhaps, is the family of differential microphone arrays (DMAs) that have the promise to form frequency-independent responses. Moreover, they have the potential to attain high directional gains with small and compact apertures. As a result, this type of microphone arrays has drawn much research and development attention recently. This book is intended to provide a systematic study of DMAs from a signal processing perspective. The primary obj...

  16. Array Processing for Radar: Achievements and Challenges

    Directory of Open Access Journals (Sweden)

    Ulrich Nickel

    2013-01-01

    Full Text Available Array processing for radar is well established in the literature, but only few of these algorithms have been implemented in real systems. The reason may be that the impact of these algorithms on the overall system must be well understood. For a successful implementation of array processing methods exploiting the full potential, the desired radar task has to be considered and all processing necessary for this task has to be eventually adapted. In this tutorial paper, we point out several viewpoints which are relevant in this context: the restrictions and the potential provided by different array configurations, the predictability of the transmission function of the array, the constraints for adaptive beamforming, the inclusion of monopulse, detection and tracking into the adaptive beamforming concept, and the assessment of superresolution methods with respect to their application in a radar system. The problems and achieved results are illustrated by examples from previous publications.

  17. Monolithic Time Delay Integrated APD Arrays Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The overall goal of the proposed program by Epitaxial Technologies is to develop monolithic time delay integrated avalanche photodiode (APD) arrays with sensitivity...

  18. Thin Flexible IMM Solar Array, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Thin, flexible, and highly efficient solar arrays are needed that package compactly for launch and deploy into large, structurally stable high power generators....

  19. Integrated Spatial Filter Array, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — To address the NASA Earth Science Division need for spatial filter arrays for amplitude and wavefront control, Luminit proposes to develop a novel Integrated Spatial...

  20. Stretched Lens Array Photovoltaic Concentrator Technology Developed

    Science.gov (United States)

    Piszczor, Michael F., Jr.; O'Neill, Mark J.

    2004-01-01

    Solar arrays have been and continue to be the mainstay in providing power to nearly all commercial and government spacecraft. Light from the Sun is directly converted into electrical energy using solar cells. One way to reduce the cost of future space power systems is by minimizing the size and number of expensive solar cells by focusing the sunlight onto smaller cells using concentrator optics. The stretched lens array (SLA) is a unique concept that uses arched Fresnel lens concentrators to focus sunlight onto a line of high-efficiency solar cells located directly beneath. The SLA concept is based on the Solar Concentrator Array with Refractive Linear Element Technology (SCARLET) design that was used on NASA's New Millennium Deep Space 1 mission. The highly successful asteroid/comet rendezvous mission (1998 to 2001) demonstrated the performance and long-term durability of the SCARLET/SLA solar array design and set the foundation for further improvements to optimize its performance.

  1. Spatiotemporal light localization in infiltrated waveguide arrays

    DEFF Research Database (Denmark)

    Rasmussen, Per Dalgaard; Neshev, D.N-; Sukhorukov, A.A.

    2008-01-01

    We study light propagation in hexagonal waveguide arrays and show that simultaneous spatiotemporal localisation is possible by combination of engineered anomalous dispersion through selective excitation of Bloch-modes and spatial confinement in a nonlinear defect mode....

  2. Ferrite LTCC based phased array antennas

    KAUST Repository

    Ghaffar, Farhan A.

    2016-11-02

    Two phased array antennas realized in multilayer ferrite LTCC technology are presented in this paper. The use of embedded bias windings in these designs allows the negation of external magnets which are conventionally employed with bulk ferrite medium. This reduces the required magnetostatic field strength by 90% as compared to the traditional designs. The phase shifters are implemented using the SIW technology. One of the designs is operated in the half mode waveguide topology while the other design is based on standard full mode waveguide operation. The two phase shifter designs are integrated with two element patch antenna array and slotted SIW array respectively. The array designs demonstrate a beam steering of 30° and ±19° respectively for a current excitation of 200 mA. The designs, due to their small factor can be easily integrated in modern communication systems which is not possible in the case of bulk ferrite based designs.

  3. Density controlled carbon nanotube array electrodes

    Science.gov (United States)

    Ren, Zhifeng F [Newton, MA; Tu, Yi [Belmont, MA

    2008-12-16

    CNT materials comprising aligned carbon nanotubes (CNTs) with pre-determined site densities, catalyst substrate materials for obtaining them and methods for forming aligned CNTs with controllable densities on such catalyst substrate materials are described. The fabrication of films comprising site-density controlled vertically aligned CNT arrays of the invention with variable field emission characteristics, whereby the field emission properties of the films are controlled by independently varying the length of CNTs in the aligned array within the film or by independently varying inter-tubule spacing of the CNTs within the array (site density) are disclosed. The fabrication of microelectrode arrays (MEAs) formed utilizing the carbon nanotube material of the invention is also described.

  4. Fiber Optic Pressure Sensor Array, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — VIP Sensors proposes to develop a Fiber Optic Pressure Sensor Array System for measuring air flow pressure at multiple points on the skin of aircrafts for Flight...

  5. SMEX-Lite Modular Solar Array Architecture

    Science.gov (United States)

    Lyons, John

    2002-01-01

    For the most part, Goddard solar arrays have been custom designs that are unique to each mission. The solar panel design has been frozen prior to issuing an RFP for their procurement. There has typically been 6-9 months between RFP release and contract award, followed by an additional 24 months for performance of the contract. For Small Explorer (SMEX) missions, with three years between mission definition and launch, this has been a significant problem. The SMEX solar panels have been sufficiently small that the contract performance period has been reduced to 12-15 months. The bulk of this time is used up in the final design definition and fabrication of flight solar cell assemblies. Even so, it has been virtually impossible to have the spacecraft design at a level of maturity sufficient to freeze the solar panel geometry and release the RFP in time to avoid schedule problems with integrating the solar panels to the spacecraft. With that in mind, the SMEX-Lite project team developed a modular architecture for the assembly of solar arrays to greatly reduce the cost and schedule associated with the development of a mission- specific solar array. In the modular architecture, solar cells are fabricated onto small substrate panels. This modular panel (approximately 8.5" x 17" in this case) becomes the building block for constructing solar arrays for multiple missions with varying power requirements and geometrical arrangements. The mechanical framework that holds these modules together as a solar array is the only mission-unique design, changing in size and shape as required for each mission. There are several advantages to this approach. First, the typical solar array development cycle requires a mission unique design, procurement, and qualification including a custom qualification panel. With the modular architecture, a single qualification of the SMEX-Lite modules and the associated mechanical framework in a typical configuration provided a qualification by

  6. Digital Array Gas Radiometer (DAGR) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The digital array gas radiometer (DAGR) is a new sensor design for accurate measurement and monitoring of trace gases in the boundary layer from space, aircraft, or...

  7. Transition edge sensor series array bolometer

    International Nuclear Information System (INIS)

    Beyer, J

    2010-01-01

    A transition edge sensor series array (TES-SA) is an array of identical TESs that are connected in series by low-inductance superconducting wiring. The array elements are equally and well thermally coupled to the absorber and respond to changes in the absorber temperature in synchronization. The TES-SA total resistance increases compared to a single TES while the shape of the superconducting transition is preserved. We are developing a TES-SA with a large number, hundreds to thousands, of array elements with the goal of enabling the readout of a TES-based bolometer operated at 4.2 K with a semiconductor-based amplifier located at room temperature. The noise and dynamic performance of a TES-SA bolometer based on a niobium/aluminum bilayer is analyzed. It is shown that stable readout of the bolometer with a low-noise transimpedance amplifier is feasible.

  8. Applications of the phased array technique

    International Nuclear Information System (INIS)

    Erhard, A.; Schenk, G.; Hauser, Th.; Voelz, U.

    1999-01-01

    The application of the phased array technique was limited to heavy and thick wall components as present in the nuclear industry. With the improvement of the equipment and probes other application areas are now open for the phased array technique, e.g. the inspection of the turbine blade root, weld inspection in a wall thickness range between 12 and 40 mm, inspection of aircraft components, inspection of spot welds or inspection of concretes. The aim of the use of phased array techniques has not been changed related to the first applications, i.e. the adaptation of the sound beam to the geometry by steering the angel of incidence or the skewing angle as well as the focussing of sound fields. Due to the fact, that the new applications of the phased array techniques in some cases don't leave the laboratories for the time being, the examples of this contribution will focus applications with practical background. (orig.)

  9. CubeSat Modular Deployable Solar Arrays

    Data.gov (United States)

    National Aeronautics and Space Administration — Advanced CubeSat missions have demanding power requirements relative to their limited available surface area that drive the need for deployable solar arrays. Vendors...

  10. NASA Solar Array Demonstrates Commercial Potential

    Science.gov (United States)

    Creech, Gray

    2006-01-01

    A state-of-the-art solar-panel array demonstration site at NASA's Dryden Flight Research Center provides a unique opportunity for studying the latest in high-efficiency solar photovoltaic cells. This five-kilowatt solar-array site (see Figure 1) is a technology-transfer and commercialization success for NASA. Among the solar cells at this site are cells of a type that was developed in Dryden Flight Research Center s Environmental Research Aircraft and Sensor Technology (ERAST) program for use in NASA s Helios solar-powered airplane. This cell type, now denoted as A-300, has since been transferred to SunPower Corporation of Sunnyvale, California, enabling mass production of the cells for the commercial market. High efficiency separates these advanced cells from typical previously commercially available solar cells: Whereas typical previously commercially available cells are 12 to 15 percent efficient at converting sunlight to electricity, these advanced cells exhibit efficiencies approaching 23 percent. The increase in efficiency is due largely to the routing of electrical connections behind the cells (see Figure 2). This approach to increasing efficiency originated as a solution to the problem of maximizing the degree of utilization of the limited space available atop the wing of the Helios airplane. In retrospect, the solar cells in use at this site could be used on Helios, but the best cells otherwise commercially available could not be so used, because of their lower efficiencies. Historically, solar cells have been fabricated by use of methods that are common in the semiconductor industry. One of these methods includes the use of photolithography to define the rear electrical-contact features - diffusions, contact openings, and fingers. SunPower uses these methods to produce the advanced cells. To reduce fabrication costs, SunPower continues to explore new methods to define the rear electrical-contact features. The equipment at the demonstration site includes

  11. A Host-Based RT-PCR Gene Expression Signature to Identify Acute Respiratory Viral Infection

    Science.gov (United States)

    Zaas, Aimee K.; Burke, Thomas; Chen, Minhua; McClain, Micah; Nicholson, Bradly; Veldman, Timothy; Tsalik, Ephraim L.; Fowler, Vance; Rivers, Emanuel P.; Otero, Ronny; Kingsmore, Stephen F.; Voora, Deepak; Lucas, Joseph; Hero, Alfred O.; Carin, Lawrence; Woods, Christopher W.; Ginsburg, Geoffrey S.

    2014-01-01

    Improved ways to diagnose acute respiratory viral infections could decrease inappropriate antibacterial use and serve as a vital triage mechanism in the event of a potential viral pandemic. Measurement of the host response to infection is an alternative to pathogen-based diagnostic testing and may improve diagnostic accuracy. We have developed a host-based assay with a reverse transcription polymerase chain reaction (RT-PCR) TaqMan low-density array (TLDA) platform for classifying respiratory viral infection. We developed the assay using two cohorts experimentally infected with influenza A H3N2/Wisconsin or influenza A H1N1/Brisbane, and validated the assay in a sample of adults presenting to the emergency department with fever (n = 102) and in healthy volunteers (n = 41). Peripheral blood RNA samples were obtained from individuals who underwent experimental viral challenge or who presented to the emergency department and had microbiologically proven viral respiratory infection or systemic bacterial infection. The selected gene set on the RT-PCR TLDA assay classified participants with experimentally induced influenza H3N2 and H1N1 infection with 100 and 87% accuracy, respectively. We validated this host gene expression signature in a cohort of 102 individuals arriving at the emergency department. The sensitivity of the RT-PCR test was 89% [95% confidence interval (CI), 72 to 98%], and the specificity was 94% (95% CI, 86 to 99%). These results show that RT-PCR–based detection of a host gene expression signature can classify individuals with respiratory viral infection and sets the stage for prospective evaluation of this diagnostic approach in a clinical setting. PMID:24048524

  12. Accurate measurement of transgene copy number in crop plants using droplet digital PCR.

    Science.gov (United States)

    Collier, Ray; Dasgupta, Kasturi; Xing, Yan-Ping; Hernandez, Bryan Tarape; Shao, Min; Rohozinski, Dominica; Kovak, Emma; Lin, Jeanie; de Oliveira, Maria Luiza P; Stover, Ed; McCue, Kent F; Harmon, Frank G; Blechl, Ann; Thomson, James G; Thilmony, Roger

    2017-06-01

    Genetic transformation is a powerful means for the improvement of crop plants, but requires labor- and resource-intensive methods. An efficient method for identifying single-copy transgene insertion events from a population of independent transgenic lines is desirable. Currently, transgene copy number is estimated by either Southern blot hybridization analyses or quantitative polymerase chain reaction (qPCR) experiments. Southern hybridization is a convincing and reliable method, but it also is expensive, time-consuming and often requires a large amount of genomic DNA and radioactively labeled probes. Alternatively, qPCR requires less DNA and is potentially simpler to perform, but its results can lack the accuracy and precision needed to confidently distinguish between one- and two-copy events in transgenic plants with large genomes. To address this need, we developed a droplet digital PCR-based method for transgene copy number measurement in an array of crops: rice, citrus, potato, maize, tomato and wheat. The method utilizes specific primers to amplify target transgenes, and endogenous reference genes in a single duplexed reaction containing thousands of droplets. Endpoint amplicon production in the droplets is detected and quantified using sequence-specific fluorescently labeled probes. The results demonstrate that this approach can generate confident copy number measurements in independent transgenic lines in these crop species. This method and the compendium of probes and primers will be a useful resource for the plant research community, enabling the simple and accurate determination of transgene copy number in these six important crop species. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

  13. Frequency Domain Electromagnetic Sensor Array Development

    Science.gov (United States)

    2007-09-17

    undergone limited bench testing. The design is based on the GEM-3 active primary field cancellation technology, which creates a "magnetic cavity" for...responses at three receiver locations. ............................................... 13 Figure 16. Warmup drift for the GEM-3 array at the Tx2-2... engineering details for application to single sensor and towed array configurations have been worked out in ESTCP projects MM-0208, -0414 and - 0531

  14. Miniaturized Retrodirective Arrays for a Nanosatellite Platform

    Science.gov (United States)

    2012-01-01

    Miniaturized Retrodirective Arrays for a Nanosatellite Platform NIA Sb. GRANT NUMBER N00014-1 0-1-0056 Sc. PROGRAM ELEMENT NUMBER . NIA 6. AUTHOR(S) Sd...into a mobile platfonn, such as a nanosatellite , presents unique challenges due to the platform’s restrictions in terms of size, weight, and power...of power detection and null scanning. 1S. SUBJECT TERMS Retrodirective Array, Nanosatellite 16. SECURITY CLASSIFICATION OF: a. REPORT b. ABSTRACT

  15. What Else Is Decidable about Integer Arrays?

    OpenAIRE

    Habermehl, Peter; Iosif, Radu; Vojnar, Tomáš

    2008-01-01

    International audience; We introduce a new decidable logic for reasoning about infinite arrays of integers. The logic is in the ∃ * ∀ * first-order fragment and allows (1) Presburger constraints on existentially quantified variables, (2) difference constraints as well as periodicity constraints on universally quantified indices, and (3) difference constraints on values. In particular, using our logic, one can express constraints on consecutive elements of arrays (e.g. ∀i. 0 ≤ i < n → a[i + 1]...

  16. High Efficiency mm-Wave Transmitter Array

    Science.gov (United States)

    2016-09-01

    Research Triangle Park, NC 27709-2211 Mm-waves. power amplifiers, Si integrated circuits, antenna arrays REPORT DOCUMENTATION PAGE 11. SPONSOR...Array Report Title High efficiency, high power transmitters integrated in silicon at 45, 94 and 138 GHz were developed. Our approach employs CMOS-SOI...based power amplifiers had a variety of advantages over nMOS PAs. Advancements have been reported in detail in numerous publications (13 journal

  17. Thermophotovoltaic Arrays for Electrical Power Generation

    Energy Technology Data Exchange (ETDEWEB)

    Sarnoff Corporation

    2003-11-18

    Sarnoff has designed an integrated array of thermophotovoltaic (TPV) cells based on the In(Al)GaAsSb/GaSb materials system. These arrays will be used in a system to generate electrical power from a radioisotope heat source that radiates at temperatures from 700 to 1000 C. Two arrays sandwich the slab heat source and will be connected in series to build voltage. Between the arrays and the heat source is a spectral control filter that transmits above-bandgap radiation and reflects below-bandgap radiation. The goal is to generate 5 mW of electrical power at 3 V from a 700 C radiant source. Sarnoff is a leader in antimonide-based TPV cell development. InGaAsSb cells with a bandgap of 0.53 eV have operated at system conversion efficiencies greater than 17%. The system included a front-surface filter, and a 905 C radiation source. The cells were grown via organo-metallic vapor-phase epitaxy. Sarnoff will bring this experience to bear on the proposed project. The authors first describe array and cell architecture. They then present calculated results showing that about 80 mW of power can be obtained from a 700 C radiator. Using a conservative array design, a 5-V output is possible.

  18. 2D array based on fermat spiral

    Science.gov (United States)

    Martínez, O.; Martín, C. J.; Godoy, G.; Ullate, L. G.

    2010-01-01

    The main challenge faced by 3D ultrasonic imaging with 2D array transducer is the large number of elements required to achieve an acceptable level of quality in the images. Therefore, the optimization of the array layout to reduce the number of active elements in the aperture has been a research topic in the last years. Nowadays, CMUT array technology has made viable the production of 2D arrays with larger flexibility on elements size, shape and position. This is opening new options in 2D array design, allowing to revise as viable alternatives others layouts that had been studied in the past, like circular and Archimedes spiral layout. In this work the problem of designing an imaging system array with a diameter of 60 λ and a limited number of elements using the Fermat spiral layout has been studied. This study has been done for two different numbers of electronic channels (N = 128 and N = 256). As summary, a general discussion of the results and the most interesting cases are presented.

  19. Tunable nanoparticle arrays at charged interfaces.

    Science.gov (United States)

    Srivastava, Sunita; Nykypanchuk, Dmytro; Fukuto, Masafumi; Gang, Oleg

    2014-10-28

    Structurally tunable two-dimensional (2D) arrays of nanoscale objects are important for modulating functional responses of thin films. We demonstrate that such tunable and ordered nanoparticles (NP) arrays can be assembled at charged air-water interfaces from nanoparticles coated with polyelectrolyte chains, DNA. The electrostatic attraction between the negatively charged nonhybridizing DNA-coated gold NPs and a positively charged lipid layer at the interface facilitates the formation of a 2D hexagonally closed packed (HCP) nanoparticle lattice. We observed about 4-fold change of the monolayer nanoparticle density by varying the ionic strength of the subphase. The tunable NP arrays retain their structure reasonably well when transferred to a solid support. The influence of particle's DNA corona and lipid layer composition on the salt-induced in-plane and normal structural evolution of NP arrays was studied in detail using a combination of synchrotron-based in situ surface scattering methods, grazing incidence X-ray scattering (GISAXS), and X-ray reflectivity (XRR). Comparative analysis of the interparticle distances as a function of ionic strength reveals the difference between the studied 2D nanoparticle arrays and analogous bulk polyelectrolyte star polymers systems, typically described by Daoud-Cotton model and power law scaling. The observed behavior of the 2D nanoparticle array manifests a nonuniform deformation of the nanoparticle DNA corona due to its electrostatically induced confinement at the lipid interface. The present study provides insight on the interfacial properties of the NPs coated with charged soft shells.

  20. Terahertz Array Receivers with Integrated Antennas

    Science.gov (United States)

    Chattopadhyay, Goutam; Llombart, Nuria; Lee, Choonsup; Jung, Cecile; Lin, Robert; Cooper, Ken B.; Reck, Theodore; Siles, Jose; Schlecht, Erich; Peralta, Alessandro; hide

    2011-01-01

    Highly sensitive terahertz heterodyne receivers have been mostly single-pixel. However, now there is a real need of multi-pixel array receivers at these frequencies driven by the science and instrument requirements. In this paper we explore various receiver font-end and antenna architectures for use in multi-pixel integrated arrays at terahertz frequencies. Development of wafer-level integrated terahertz receiver front-end by using advanced semiconductor fabrication technologies has progressed very well over the past few years. Novel stacking of micro-machined silicon wafers which allows for the 3-dimensional integration of various terahertz receiver components in extremely small packages has made it possible to design multi-pixel heterodyne arrays. One of the critical technologies to achieve fully integrated system is the antenna arrays compatible with the receiver array architecture. In this paper we explore different receiver and antenna architectures for multi-pixel heterodyne and direct detector arrays for various applications such as multi-pixel high resolution spectrometer and imaging radar at terahertz frequencies.

  1. Si Wire-Array Solar Cells

    Science.gov (United States)

    Boettcher, Shannon

    2010-03-01

    Micron-scale Si wire arrays are three-dimensional photovoltaic absorbers that enable orthogonalization of light absorption and carrier collection and hence allow for the utilization of relatively impure Si in efficient solar cell designs. The wire arrays are grown by a vapor-liquid-solid-catalyzed process on a crystalline (111) Si wafer lithographically patterned with an array of metal catalyst particles. Following growth, such arrays can be embedded in polymethyldisiloxane (PDMS) and then peeled from the template growth substrate. The result is an unusual photovoltaic material: a flexible, bendable, wafer-thickness crystalline Si absorber. In this paper I will describe: 1. the growth of high-quality Si wires with controllable doping and the evaluation of their photovoltaic energy-conversion performance using a test electrolyte that forms a rectifying conformal semiconductor-liquid contact 2. the observation of enhanced absorption in wire arrays exceeding the conventional light trapping limits for planar Si cells of equivalent material thickness and 3. single-wire and large-area solid-state Si wire-array solar cell results obtained to date with directions for future cell designs based on optical and device physics. In collaboration with Michael Kelzenberg, Morgan Putnam, Joshua Spurgeon, Daniel Turner-Evans, Emily Warren, Nathan Lewis, and Harry Atwater, California Institute of Technology.

  2. Construction of a recombinant plasmid as reaction control in routine PCR for detection of contagious equine metritis (CEM-PCR).

    Science.gov (United States)

    Niwa, Hidekazu; Anzai, Toru; Hobo, Seiji

    2007-11-01

    Contagious equine metritis (CEM) is a highly contagious bacterial venereal disease of horses caused by Taylorella equigenitalis. CEM-PCR is a semi-nested PCR method for detecting this bacterium. Although this technique is regarded as a sensitive diagnostic method for CEM, there are risks of it generating false positive and false negative results. In this study, we constructed a recombinant plasmid (CEM-POS) as reaction control to assure adequate PCR reaction and prevent false positive results caused by contamination of the reaction control in routine CEM-PCR examinations. CEM-POS was constructed by insertion of rpoB fragments from Rhodococcus equi into CEM-1P, which is a recombinant plasmid that includes a T. equigenitalis-specific sequence region. In CEM-PCR, the size of the PCR product from CEM-POS was clearly different from the true positive PCR product. In addition, CEM-POS retained high stability under convenient storage conditions of 4 degrees C. These results suggest CEM-POS to be a useful tool as a reaction control in routine CEM-PCR examinations.

  3. RT-PCR and real-time PCR analysis of E2A-PBX1, TEL-AML1 ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Genetics; Volume 90; Issue 2. RT-PCR and real-time PCR analysis of E2A-PBX1, TEL-AML1, mBCR-ABL and MLL-AF4 fusion gene transcripts in de novo B-lineage acute lymphoblastic leukaemia patients in south India. Natarajan Sudhakar Kamalalayam Raghavan Rajalekshmy Thangarajan ...

  4. Comparison of droplet digital PCR and seminested real-time PCR for quantification of cell-associated HIV-1 RNA

    NARCIS (Netherlands)

    Kiselinova, Maja; Pasternak, Alexander O.; de Spiegelaere, Ward; Vogelaers, Dirk; Berkhout, Ben; Vandekerckhove, Linos

    2014-01-01

    Cell-associated (CA) HIV-1 RNA is considered a potential marker for assessment of viral reservoir dynamics and antiretroviral therapy (ART) response in HIV-infected patients. Recent studies employed sensitive seminested real-time quantitative (q)PCR to quantify CA HIV-1 RNA. Digital PCR has been

  5. Critical appraisal of quantitative PCR results in colorectal cancer research: Can we rely on published qPCR results?

    NARCIS (Netherlands)

    Dijkstra, J.R.; Kempen, L.C.L.T. van; Nagtegaal, I.D.; Bustin, S.A.

    2014-01-01

    The use of real-time quantitative polymerase chain reaction (qPCR) in cancer research has become ubiquitous. The relative simplicity of qPCR experiments, which deliver fast and cost-effective results, means that each year an increasing number of papers utilizing this technique are being published.

  6. Development of an Oligonucleotide Array for Direct Detection of Fungi in Sputum Samples from Patients with Cystic Fibrosis▿ †

    Science.gov (United States)

    Bouchara, Jean-Phillippe; Hsieh, Hsin Yi; Croquefer, Sabine; Barton, Richard; Marchais, Veronique; Pihet, Marc; Chang, Tsung Chain

    2009-01-01

    Cystic fibrosis (CF) is the most common inherited genetic disease in Caucasian populations. Besides bacteria, many species of fungi may colonize the respiratory tract of these patients, sometimes leading to true respiratory infections. In this study, an oligonucleotide array capable of identifying 20 fungal species was developed to directly detect fungi in the sputum samples of CF patients. Species-specific oligonucleotide probes were designed from the internal transcribed spacer (ITS) regions of the rRNA operon and immobilized on a nylon membrane. The fungal ITS regions were amplified by PCR and hybridized to the array for species identification. The array was validated by testing 182 target strains (strains which we aimed to identify) and 141 nontarget strains (135 species), and a sensitivity of 100% and a specificity of 99.2% were obtained. The validated array was then used for direct detection of fungi in 57 sputum samples from 39 CF patients, and the results were compared to those obtained by culture. For 16 sputum samples, the results obtained by the array corresponded with those obtained by culture. For 33 samples, the array detected more fungal species than culture did, while the reverse was found for eight samples. The accuracy of the array for fungal detection in sputum samples was confirmed (or partially confirmed) in some samples by cloning and resequencing the amplified ITS fragments. The present array is a useful tool for both the simultaneous detection of multiple fungal species present in the sputa of CF patients and the identification of fungi isolated from these patients. PMID:19020057

  7. Transient neonatal diabetes mellitus with macroglossia diagnosed by methylation specific PCR (MS-PCR

    Directory of Open Access Journals (Sweden)

    Hye Young Jin

    2010-03-01

    Full Text Available Transient neonatal diabetes mellitus (TNDM has been associated with paternal uniparental isodisomy of chromosome 6, paternally inherited duplication of 6q24, or a methylation defect at a CpG island of the ZAC or HYMAI gene. We experienced a case of TNDM in which the patient presented with hyperglycemia, macroglossia, and intrauterine growth retardation, caused by a paternally derived HYMAI. An 18-day-old female infant was admitted to the hospital because of macroglossia and recurrent hyperglycemia. In addition to the macroglossia, she also presented with large fontanelles, micrognathia, and prominent eyes. Serum glucose levels were 200&#8211;300 mg/dL and they improved spontaneously 2 days after admission. To identify the presence of a maternal methylated allele, bisulfite-treated genomic DNA from peripheral blood was prepared and digested with BssHII after polymerase chain reaction (PCR amplification with methylation-specific HYMAI primers. PCR and restriction fragment length polymorphism analysis showed that the patient had only the paternal origin of the HYMA1 gene. TNDM is associated with a methylation defect in chromosome 6, suggesting that an imprinted gene on chromosome 6 is responsible for this phenotype.

  8. Aspects of the Subarrayed Array Processing for the Phased Array Radar

    Directory of Open Access Journals (Sweden)

    Hang Hu

    2015-01-01

    Full Text Available This paper gives an overview on the research status, developments, and achievements of subarrayed array processing for the multifunction phased array radar. We address some issues concerning subarrayed adaptive beamforming, subarrayed fast-time space-time adaptive processing, subarray-based sidelobe reduction of sum and difference beam, subarrayed adapted monopulse, subarrayed superresolution direction finding, subarray configuration optimization in ECCM (electronic counter-countermeasure, and subarrayed array processing for MIMO-PAR. In this review, several viewpoints relevant to subarrayed array processing are pointed out and the achieved results are demonstrated by numerical examples.

  9. Simultaneous detection and differentiation of human papillomavirus genotypes 6, 11, 16 and 18 by AllGlo quadruplex quantitative PCR.

    Directory of Open Access Journals (Sweden)

    Daojun Yu

    Full Text Available BACKGROUND: Human papillomaviruses (HPV are classified into high-risk HPV and low-risk HPV. The most common high-risk HPV types in cervical cancer are HPV 16 and 18, and the most common low-risk types causing genital warts are HPV 6 and HPV 11. In this study, applying novel AllGlo fluorescent probes, we established a quadruplex quantitative PCR method to simultaneously detect and differentiate HPV 6, 11, 16 and 18 in a single tube. METHODS: The specificity, the sensitivity, the detection limit, the reproducibility and the standard curve of this method were examined. Finally, clinical samples that had been tested previously by TaqMan PCR and HPV GenoArray (GA test were used to verify the accuracy and sensitivity of the method. RESULTS: The assay has a sensitivity of 10(1 to 10(2 copies/test and a linear detection range from 10(1 to 10(8 copies/test. The mean amplification efficiencies for HPV 6, 11, 16, and 18 were 0.97, 1.10, 0.93 and 1.20, respectively, and the mean correlation coefficient (r(2 of each standard curve was above 0.99 for plasmid templates ranging from 10(3 to 10(7 copies/test. There was 100% agreement between the AllGlo quadruplex quantitative PCR, HPV GA test and TaqMan uniplex qPCR methods. CONCLUSIONS: AllGlo quadruplex quantitative PCR in a single tube has the advantages of relatively high throughput, good reproducibility, high sensitivity, high specificity, and a wide linear range of detection. The convenient single tube format makes this assay a powerful tool for the studies of mixed infections by multiple pathogens, viral typing and viral load quantification.

  10. Comparison of PCR-DGGE and Nested-PCR-DGGE Approach for Ammonia Oxidizers Monitoring in Membrane Bioreactors’ Activated Sludge

    Directory of Open Access Journals (Sweden)

    Ziembińska-Buczyńska Aleksandra

    2014-12-01

    Full Text Available Nitritation, the first stage of ammonia removal process is known to be limiting for total process performance. Ammonia oxidizing bacteria (AOB which perform this process are obligatory activated sludge habitants, a mixture consisting of Bacteria, Protozoa and Metazoa used for biological wastewater treatment. Due to this fact they are an interesting bacterial group, from both the technological and ecological point of view. AOB changeability and biodiversity analyses both in wastewater treatment plants and lab-scale reactors are performed on the basis of 16S rRNA gene sequences using PCR-DGGE (Polymerase Chain Reaction – Denaturing Gradient Gel Electrophoresis as a molecular biology tool. AOB researches are usually led with nested PCR. Because the application of nested PCR is laborious and time consuming, we have attempted to check the possibility of using only first PCR round to obtain DGGE fingerprinting of microbial communities. In this work we are comparing the nested and non-nested PCR-DGGE monitoring of an AOB community and presenting advantages and disadvantages of both methods used. The experiment revealed that PCR technique is a very sensitive tool for the amplification of even a minute amount of DNA sample. But in the case of nested-PCR, the sensitivity is higher and the template amount could be even smaller. The nested PCR-DGGE seems to be a better tool for AOB community monitoring and complexity research in activated sludge, despite shorter fragments of DNA amplification which seems to be a disadvantage in the case of bacteria identification. It is recommended that the sort of analysis approach should be chosen according to the aim of the study: nested-PCR-DGGE for community complexity analysis, while PCR-DGGE for identification of the dominant bacteria.

  11. Multiplex PCR system for the rapid diagnosis of respiratory virus infection: systematic review and meta-analysis.

    Science.gov (United States)

    Huang, H-S; Tsai, C-L; Chang, J; Hsu, T-C; Lin, S; Lee, C-C

    2017-12-05

    To provide a summary of evidence for the diagnostic accuracies of three multiplex PCR systems (mPCRs)-BioFire FilmArray RP (FilmArray), Nanosphere Verigene RV+ test (Verigene RV+) and Hologic Gen-Probe Prodesse assays-on the detection of viral respiratory infections. A comprehensive search up to 1 July 2017 was conducted on Medline and Embase for studies that utilized FilmArray, Verigene RV+ and Prodesse for diagnosis of viral respiratory infections. A summary of diagnostic accuracies for the following five viruses were calculated: influenza A virus (FluA), influenza B virus, respiratory syncytial virus, human metapneumovirus and adenovirus. Hierarchical summary receiver operating curves were used for estimating the viral detection performance per assay. Twenty studies of 5510 patient samples were eligible for analysis. Multiplex PCRs demonstrated high diagnostic accuracy, with area under the receiver operating characteristic curve (AUROC) equal to or more than 0.98 for all the above viruses except for adenovirus (AUROC 0.89). FilmArray, Verigene RV+ and ProFlu+ (the only Prodesse assay with enough data) demonstrated a summary sensitivity for FluA of 0.911 (95% confidence interval, 0.848-0.949), 0.949 (95% confidence interval, 0.882-0.979) and 0.954 (95% confidence interval, 0.871-0.985), respectively. The three mPCRs were comparable in terms of detection of FluA. Point estimates calculated from eligible studies showed that the three mPCRs (FilmArray, Verigene RV+ and ProFlu+) are highly accurate and may provide important diagnostic information for early identification of respiratory virus infections. In patients with low pretest probability for FluA, these three mPCRs can predict a low possibility of infection and may justify withholding empirical antiviral treatments. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  12. Evaluation of fliC-d based direct blood PCR assays for typhoid diagnosis

    OpenAIRE

    Das, Surojit; Ray, Ujjwayini; Akhter, Irfaan; Chattopadhyay, Arka; Paul, Dilip Kumar; Dutta, Shanta

    2016-01-01

    Background Typhoid cases need to be diagnosed accurately for early antibiotic therapy and reducing mortality. Identification of Salmonella Typhi (S. Typhi) in blood culture is conclusive, but has poor sensitivity. Detection of S. Typhi by PCR from blood sample has shown promise. Real-time quantitative PCR (Q-PCR) has been widely used in diagnostics for its rapidity and reliability. In the present study, the performance of molecular methods like conventional PCR (C-PCR), nested PCR (N-PCR) and...

  13. RT-PCR Protocols - Methods in Molecular Biology

    Directory of Open Access Journals (Sweden)

    Manuela Monti

    2011-03-01

    Full Text Available “The first record I have of it, is when I made a computer file which I usually did whenever I had an idea, that would have been on the Monday when I got back, and I called it Chain Reaction.POL, meaning polymerase. That was the identifier for it and later I called the thing the Polymerase Chain Reaction, which a lot of people thought was a dumb name for it, but it stuck, and it became PCR”. With these words the Nobel prize winner, Kary Mullis, explains how he named the PCR: one of the most important techniques ever invented and currently used in molecular biology. This book “RT-PCR Protocols” covers a wide range of aspects important for the setting of a PCR experiment for both beginners and advanced users. In my opinion the book is very well structured in three different sections. The first one describes the different technologies now available, like competitive RT-PCR, nested RT-PCR or RT-PCR for cloning. An important part regards the usage of PCR in single cell mouse embryos, stressing how important...........

  14. Broad-range PCR: past, present, or future of bacteriology?

    Science.gov (United States)

    Renvoisé, A; Brossier, F; Sougakoff, W; Jarlier, V; Aubry, A

    2013-08-01

    PCR targeting the gene encoding 16S ribosomal RNA (commonly named broad-range PCR or 16S PCR) has been used for 20 years as a polyvalent tool to study prokaryotes. Broad-range PCR was first used as a taxonomic tool, then in clinical microbiology. We will describe the use of broad-range PCR in clinical microbiology. The first application was identification of bacterial strains obtained by culture but whose phenotypic or proteomic identification remained difficult or impossible. This changed bacterial taxonomy and allowed discovering many new species. The second application of broad-range PCR in clinical microbiology is the detection of bacterial DNA from clinical samples; we will review the clinical settings in which the technique proved useful (such as endocarditis) and those in which it did not (such as characterization of bacteria in ascites, in cirrhotic patients). This technique allowed identifying the etiological agents for several diseases, such as Whipple disease. This review is a synthesis of data concerning the applications, assets, and drawbacks of broad-range PCR in clinical microbiology. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  15. Shape based kinetic outlier detection in real-time PCR

    Directory of Open Access Journals (Sweden)

    D'Atri Mario

    2010-04-01

    Full Text Available Abstract Background Real-time PCR has recently become the technique of choice for absolute and relative nucleic acid quantification. The gold standard quantification method in real-time PCR assumes that the compared samples have similar PCR efficiency. However, many factors present in biological samples affect PCR kinetic, confounding quantification analysis. In this work we propose a new strategy to detect outlier samples, called SOD. Results Richards function was fitted on fluorescence readings to parameterize the amplification curves. There was not a significant correlation between calculated amplification parameters (plateau, slope and y-coordinate of the inflection point and the Log of input DNA demonstrating that this approach can be used to achieve a "fingerprint" for each amplification curve. To identify the outlier runs, the calculated parameters of each unknown sample were compared to those of the standard samples. When a significant underestimation of starting DNA molecules was found, due to the presence of biological inhibitors such as tannic acid, IgG or quercitin, SOD efficiently marked these amplification profiles as outliers. SOD was subsequently compared with KOD, the current approach based on PCR efficiency estimation. The data obtained showed that SOD was more sensitive than KOD, whereas SOD and KOD were equally specific. Conclusion Our results demonstrated, for the first time, that outlier detection can be based on amplification shape instead of PCR efficiency. SOD represents an improvement in real-time PCR analysis because it decreases the variance of data thus increasing the reliability of quantification.

  16. IDENTIFIKASI TIPE HLA KELAS II DENGAN TEKNIK PCR

    Directory of Open Access Journals (Sweden)

    Ervi Salwati

    2012-09-01

    Full Text Available HLA (Human Leukocyte Antigen contains a set of genes located together on the short arm of chromosome 6. These genes control immune responses, graft acceptance or rejection and tumor surveillance. These abilities have close relationship with genetic variation (occur in "many forms" or alleles that bind and present antigens to T lymphocytes. Using advanced technology and molecular biology approaches (PCR technique detection of genetic variation in the HLA region (or HLA typing has been performed based on DNA.. PCR is an in vitro technique to amplify the DNA sequence enzymatically. "Sequence Specific Primers" (SSP are designed for this PCR to obtain amplification of specific alleles or groups of alleles. The PCR products are visualized through agarose gel electrophoresis stained with ethidium bromide. The PCR technique requires small amount of whole blood (0.5 - 1 ml, gives rapid, accurate and complete result. This paper discuss identification of HLA class II typing using PCR-SSP technique and show the examples of the results.   Key words: HLA (Human Leukocyte Antigen class II, PCR (Polymerase Chain Reaction

  17. A multiplex PCR for detection of six viruses in ducks.

    Science.gov (United States)

    Wang, Yongjuan; Zhu, Shanyuan; Hong, Weiming; Wang, Anping; Zuo, Weiyong

    2017-10-01

    In this study, six pairs of specific primers that can amplify DNA fragments of different sizes were designed and synthesized according to viral protein gene sequences published in GenBank. Then, a multiplex PCR method was established for rapid detection of duck hepatitis virus 1, duck plague virus, duck Tembusu virus, muscovy duck parvovirus, muscovy duck reovirus, and duck H9N2 avian influenza virus, and achieve simple and rapid detection of viral diseases in ducks. Single PCR was used to confirm primer specificity, and PCR conditions were optimized to construct a multiplex PCR system. Specificity and sensitivity assays were also developed. The multiplex PCR was used to detect duck embryos infected with mixed viruses and those with clinically suspected diseases to verify the feasibility of the multiplex PCR. Results show that the primers can specifically amplify target fragments, without any cross-amplification with other viruses. The multiplex PCR system can amplify six DNA fragments from the pooled viral genomes and specifically detect nucleic acids of the six duck susceptible viruses when the template amount is 10 2 copies/μl. In addition, the system can be used to detect viral nucleic acids in duck embryos infected with the six common viruses. The detection results for clinical samples are consistent with those detected by single PCR. Therefore, the established multiplex PCR method can perform specific, sensitive, and high-throughput detection of six duck-infecting viruses and can be applied to clinical identification and diagnosis of viral infection in ducks. Copyright © 2017. Published by Elsevier B.V.

  18. Phased-array radar for airborne systems

    Science.gov (United States)

    Tahim, Raghbir S.; Foshee, James J.; Chang, Kai

    2003-09-01

    Phased array antenna systems, which support high pulse rates and high transmit power, are well suited for radar and large-scale surveillance. Sensors and communication systems can function as the eyes and ears for ballistic missile defense applications, providing early warning of attack, target detection and identification, target tracking, and countermeasure decision. In such applications, active array radar systems that contain solid-state transmitter sources and low-noise preamplifiers for transmission and reception are preferred over the conventional radar antennas, because the phased array radar offers the advantages of power management and efficiency, reliability, signal reception, beam steering target detection. The current phased array radar designs are very large, complex and expensive and less efficient because of high RF losses in the phase control circuits used for beam scan. Several thousands of phase shifters and drivers may be required for a single system thus making the system very complex and expensive. This paper describes the phased array radar system based on high power T/R modules, wide-band radiating planar antenna elements and very low loss wide-band phase control circuits (requiring reduced power levels) for beam scan. The phase shifter design is based on micro-strip feed lines perturbed by the proximity of voltage controlled piezoelectric transducer (PET). Measured results have shown an added insertion loss of less than 1 dB for a phase shift of 450 degrees from 2 to 20 GHz. The new wideband phased array radar design provides significant reduction in size cost and weight. Compared to the conventional phased array systems, the cost saving is more than 15 to 1.

  19. PCR IN TRAUMATOLOGY AND ORTHOPAEDICS: METHOD DESCRIPTION AND APPLICABILITY

    Directory of Open Access Journals (Sweden)

    E. M. Polyakova

    2014-01-01

    Full Text Available Review brief presents description of polymerase chain reaction method (PCR and its most common variants. Three PCR-based lines of research, carried out in the traumatology and orthopaedics, include identifying a causative agents of the implant-associated infection after orthopaedic surgery; detection of antibiotic resistance genes and biofilm forming genes. It was shown that PCR can be used as additional method for detection of genetic disorders, significant for traumatology and orthopaedics, and for investigation of cartilage and bone regeneration.

  20. PCR amplification on microarrays of gel immobilized oligonucleotides

    Science.gov (United States)

    Strizhkov, Boris; Tillib, Sergei; Mikhailovich, Vladimir; Mirzabekov, Andrei

    2003-11-04

    The invention relates two general methods for performing PCR amplification, combined with the detection and analysis of the PCR products on a microchip. In the first method, the amplification occurs both outside and within a plurality of gel pads on a microchip, with at least one oligonucleotide primer immobilized in a gel pad. In the second method, PCR amplification also takes place within gel pads on a microchip, but the pads are surrounded by a hydrophobic liquid such as that which separates the individual gel pads into environments which resemble micro-miniaturized test tubes.

  1. Diagnostic PCR: Comparative sensitivity of four probe chemistries

    DEFF Research Database (Denmark)

    Josefsen, Mathilde Hartmann; Löfström, Charlotta; Sommer, Helle Mølgaard

    2009-01-01

    Three probe chemistries: locked nucleic acid (LNA), minor groove binder (MGB) and Scorpion were compared with a TaqMan probe in a validated real-time PCR assay for detection of food-borne thermotolerant Campylobacter. The LNA probe produced significantly lower Ct-values and a higher proportion...... of positive PCR responses analyzing less than 150 DNA copies than the TaqMan probe. Choice of probe chemistry clearly has an impact on the sensitivity of PCR assays, and should be considered in an optimization strategy....

  2. Adaptive port-starboard beamforming of triplet arrays

    NARCIS (Netherlands)

    Beerens, S.P.; Been, R.; Groen, J.; Noutary, E.; Doisy, Y.

    2000-01-01

    Triplet arrays are single line arrays with three hydrophones on a circular section of the array. The triplet structure provides immediate port-starboard (PS) discrimination. This paper discusses the theoretical and experimental performance of triplet arrays. Results are obtained on detection gain

  3. Sorting white blood cells in microfabricated arrays

    Science.gov (United States)

    Castelino, Judith Andrea Rose

    Fractionating white cells in microfabricated arrays presents the potential for detecting cells with abnormal adhesive or deformation properties. A possible application is separating nucleated fetal red blood cells from maternal blood. Since fetal cells are nucleated, it is possible to extract genetic information about the fetus from them. Separating fetal cells from maternal blood would provide a low cost noninvasive prenatal diagnosis for genetic defects, which is not currently available. We present results showing that fetal cells penetrate further into our microfabricated arrays than adult cells, and that it is possible to enrich the fetal cell fraction using the arrays. We discuss modifications to the array which would result in further enrichment. Fetal cells are less adhesive and more deformable than adult white cells. To determine which properties limit penetration, we compared the penetration of granulocytes and lymphocytes in arrays with different etch depths, constriction size, constriction frequency, and with different amounts of metabolic activity. The penetration of lymphocytes and granulocytes into constrained and unconstrained arrays differed qualitatively. In constrained arrays, the cells were activated by repeated shearing, and the number of cells stuck as a function of distance fell superexponentially. In unconstrained arrays the number of cells stuck fell slower than an exponential. We attribute this result to different subpopulations of cells with different sticking parameters. We determined that penetration in unconstrained arrays was limited by metabolic processes, and that when metabolic activity was reduced penetration was limited by deformability. Fetal cells also contain a different form of hemoglobin with a higher oxygen affinity than adult hemoglobin. Deoxygenated cells are paramagnetic and are attracted to high magnetic field gradients. We describe a device which can separate cells using 10 μm magnetic wires to deflect the paramagnetic

  4. SUPERCONDUCTING QUADRUPOLE ARRAYS FOR MULTIPLE BEAM TRANSPORT

    Energy Technology Data Exchange (ETDEWEB)

    Rainer Meinke

    2003-10-01

    The goal of this research was to develop concepts for affordable, fully functional arrays of superconducting quadrupoles for multi-beam transport and focusing in heavy ion fusion (HIF)accelerators. Previous studies by the Virtual National Laboratory (VNL) collaboration have shown that the multi-beam transport system (consisting of alternating gradient quadrupole magnets, a beam vacuum system, and the beam monitor and control system) will likely be one of the most expensive and critical parts of such an accelerator. This statement is true for near-term fusion research accelerators as well as accelerators for the ultimate goal of power production via inertial fusion. For this reason, research on superconducting quadrupole arrays is both timely and important for the inertial fusion energy (IFE) research program. This research will also benefit near-term heavy ion fusion facilities such as the Integrated Research Experiment (IRE)and/or the Integrated Beam Experiment (IBX). We considered a 2-prong approach that addresses the needs of both the nearer and longer term requirements of the inertial fusion program. First, we studied the flat coil quadrupole design that was developed by LLNL; this magnet is 150 mm long with a 50 mm aperture and thus is suitable for near term experiments that require magnets of a small length to aperture ratio. Secondly, we studied the novel double-helix quadrupole (DHQ) design in a small (3 x 3) array configuration; this design can provide an important step to the longer term solution of low-cost, easy to manufacture array constructions. Our Phase I studies were performed using the AMPERES magnetostatic analysis software. Consideration of these results led to plans for future magnet R&D construction projects. The first objective of Phase I was to develop the concept of a superconducting focusing array that meets the specific requirements of a heavy ion fusion accelerator. Detailed parameter studies for such quadrupole arrays were performed

  5. Microdeletion and microduplication analysis of chinese conotruncal defects patients with targeted array comparative genomic hybridization.

    Directory of Open Access Journals (Sweden)

    Xiaohui Gong

    Full Text Available OBJECTIVE: The current study aimed to develop a reliable targeted array comparative genomic hybridization (aCGH to detect microdeletions and microduplications in congenital conotruncal defects (CTDs, especially on 22q11.2 region, and for some other chromosomal aberrations, such as 5p15-5p, 7q11.23 and 4p16.3. METHODS: Twenty-seven patients with CTDs, including 12 pulmonary atresia (PA, 10 double-outlet right ventricle (DORV, 3 transposition of great arteries (TGA, 1 tetralogy of Fallot (TOF and one ventricular septal defect (VSD, were enrolled in this study and screened for pathogenic copy number variations (CNVs, using Agilent 8 x 15K targeted aCGH. Real-time quantitative polymerase chain reaction (qPCR was performed to test the molecular results of targeted aCGH. RESULTS: Four of 27 patients (14.8% had 22q11.2 CNVs, 1 microdeletion and 3 microduplications. qPCR test confirmed the microdeletion and microduplication detected by the targeted aCGH. CONCLUSION: Chromosomal abnormalities were a well-known cause of multiple congenital anomalies (MCA. This aCGH using arrays with high-density coverage in the targeted regions can detect genomic imbalances including 22q11.2 and other 10 kinds CNVs effectively and quickly. This approach has the potential to be applied to detect aneuploidy and common microdeletion/microduplication syndromes on a single microarray.

  6. Design and operation of a portable scanner for high performance microchip capillary array electrophoresis.

    Science.gov (United States)

    Scherer, James R; Liu, Peng; Mathies, Richard A

    2010-11-01

    We have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ~20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex(®) 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification.

  7. Microsphere Suspension Array Assays for Detection and Differentiation of Hendra and Nipah Viruses

    Directory of Open Access Journals (Sweden)

    Adam J. Foord

    2013-01-01

    Full Text Available Microsphere suspension array systems enable the simultaneous fluorescent identification of multiple separate nucleotide targets in a single reaction. We have utilized commercially available oligo-tagged microspheres (Luminex MagPlex-TAG to construct and evaluate multiplexed assays for the detection and differentiation of Hendra virus (HeV and Nipah virus (NiV. Both these agents are bat-borne zoonotic paramyxoviruses of increasing concern for veterinary and human health. Assays were developed targeting multiple sites within the nucleoprotein (N and phosphoprotein (P encoding genes. The relative specificities and sensitivities of the assays were determined using reference isolates of each virus type, samples from experimentally infected horses, and archival veterinary diagnostic submissions. Results were assessed in direct comparison with an established qPCR. The microsphere array assays achieved unequivocal differentiation of HeV and NiV and the sensitivity of HeV detection was comparable to qPCR, indicating high analytical and diagnostic specificity and sensitivity.

  8. Expression Comparison of Oil Biosynthesis Genes in Oil Palm Mesocarp Tissue Using Custom Array

    Science.gov (United States)

    Wong, Yick Ching; Kwong, Qi Bin; Lee, Heng Leng; Ong, Chuang Kee; Mayes, Sean; Chew, Fook Tim; Appleton, David R.; Kulaveerasingam, Harikrishna

    2014-01-01

    Gene expression changes that occur during mesocarp development are a major research focus in oil palm research due to the economic importance of this tissue and the relatively rapid increase in lipid content to very high levels at fruit ripeness. Here, we report the development of a transcriptome-based 105,000-probe oil palm mesocarp microarray. The expression of genes involved in fatty acid (FA) and triacylglycerol (TAG) assembly, along with the tricarboxylic acid cycle (TCA) and glycolysis pathway at 16 Weeks After Anthesis (WAA) exhibited significantly higher signals compared to those obtained from a cross-species hybridization to the Arabidopsis (p-value < 0.01), and rice (p-value < 0.01) arrays. The oil palm microarray data also showed comparable correlation of expression (r2 = 0.569, p < 0.01) throughout mesocarp development to transcriptome (RNA sequencing) data, and improved correlation over quantitative real-time PCR (qPCR) (r2 = 0.721, p < 0.01) of the same RNA samples. The results confirm the advantage of the custom microarray over commercially available arrays derived from model species. We demonstrate the utility of this custom microarray to gain a better understanding of gene expression patterns in the oil palm mesocarp that may lead to increasing future oil yield. PMID:27600348

  9. Expression Comparison of Oil Biosynthesis Genes in Oil Palm Mesocarp Tissue Using Custom Array

    Directory of Open Access Journals (Sweden)

    Yick Ching Wong

    2014-11-01

    Full Text Available Gene expression changes that occur during mesocarp development are a major research focus in oil palm research due to the economic importance of this tissue and the relatively rapid increase in lipid content to very high levels at fruit ripeness. Here, we report the development of a transcriptome-based 105,000-probe oil palm mesocarp microarray. The expression of genes involved in fatty acid (FA and triacylglycerol (TAG assembly, along with the tricarboxylic acid cycle (TCA and glycolysis pathway at 16 Weeks After Anthesis (WAA exhibited significantly higher signals compared to those obtained from a cross-species hybridization to the Arabidopsis (p-value < 0.01, and rice (p-value < 0.01 arrays. The oil palm microarray data also showed comparable correlation of expression (r2 = 0.569, p < 0.01 throughout mesocarp development to transcriptome (RNA sequencing data, and improved correlation over quantitative real-time PCR (qPCR (r2 = 0.721, p < 0.01 of the same RNA samples. The results confirm the advantage of the custom microarray over commercially available arrays derived from model species. We demonstrate the utility of this custom microarray to gain a better understanding of gene expression patterns in the oil palm mesocarp that may lead to increasing future oil yield.

  10. Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.

    Directory of Open Access Journals (Sweden)

    Fabrícia Gimenes

    Full Text Available Sexually transmitted diseases (STDs may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV and genotypes by single PCR (sPCR in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%, sensitivity (100.00%, specificity (99.70%, positive (96.40% and negative predictive values (100.00% and accuracy (99.80%. The prevalence of STDs was very high (55.3%. Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks.

  11. Whole blood Nested PCR and Real-time PCR amplification of Talaromyces marneffei specific DNA for diagnosis.

    Science.gov (United States)

    Lu, Sha; Li, Xiqing; Calderone, Richard; Zhang, Jing; Ma, Jianchi; Cai, Wenying; Xi, Liyan

    2016-02-01

    Talaromyces marneffei is a dimorphic pathogenic fungus, which is a life-threatening invasive mycosis in the immunocompromised host. Prompt diagnosis of T. marneffei infection remains difficult although there has been progress in attempts to expedite the diagnosis of this infection. We previously demonstrated the value of nested polymerase chain reaction (PCR) to detect T. marneffei in paraffin embedded tissue samples with high sensitivity and specificity. In this study, this assay was used to detect the DNA of T. marneffei in whole blood samples. Real-time PCR assay was also evaluated to identify T. marneffei in the same samples. Twenty out of 30 whole blood samples (67%) collected from 23 patients were found positive by using the nested PCR assay, while 23/30 (77%) samples were found positive by using the real-time PCR assay. In order to express accurately the fungal loads, we used a normalized linearized plasmid as an internal control for real-time PCR. The assay results were correlated as the initial quantity (copies/μl) with fungal burden. These data indicate that combination of nested PCR and real-time PCR assay provides an attractive alternative for identification of T. marneffei DNA in whole blood samples of HIV-infected patients. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Designing microcapsule arrays that propagate chemical signals

    Science.gov (United States)

    Bhattacharya, Amitabh; Balazs, Anna C.

    2010-08-01

    Using analysis and simulation, we show how ordered arrays of microcapsules in solution can be harnessed to propagate chemical signals in directed and controllable ways, allowing the signals to be transmitted over macroscopic distances. The system encompasses two types of capsules that are localized on an adhesive surface. The “signaling” capsules release inducer molecules, which trigger “targets” to release nanoparticles. The released nanoparticles can bind to the underlying surface and thus, create adhesion gradients, which then propel the signaling capsules to shuttle between neighboring targets. This arrangement acts like a relay, so that triggering target capsules at a particular location in the array also triggers target capsules in adjacent locations. For an array containing two target columns, our simulations and analysis show that steady input signal leads to a sustained periodic output. For an array containing multiple target columns, we show that by introducing a prescribed ratio of nanoparticle release rates between successive target columns, a chemical signal can be propagated along the array without dissipation. We also demonstrate that similar signal transmission cannot be performed via diffusion alone.

  13. Brazilian Decimetric Array (BDA) project - Phase II

    Science.gov (United States)

    Faria, C.; Stephany, S.; Sawant, H. S.; Cecatto, J. R.; Fernandes, F. C. R.

    2010-02-01

    The configuration of the second phase of the Brazilian Decimetric Array (BDA), installed at Cachoeira Paulista, Brazil (Longitude 45° 0‧ 20″ W and Latitude 22° 41‧ 19″ S), is a T-shaped array where 21 antennas are being added to existing 5 antennas of the first phase. In the third phase, in each arm of the T array, four more antennas will be added and baselines will be increased to 2.5 × 1.25 km in east-west and south directions, respectively. The antennas will be equally spaced at the distances of 250 meters from the central antenna of the T-array. Also, the frequency range will be increased to 1.2-1.7, 2.8 and 5.6 GHz. The Second phase of the BDA should be operational by the middle of 2010 and will operate in the frequency range of (1.2-1.7) GHz for solar and non solar observations. Here, we present the characteristics of the second phase of the BDA project, details of the array configuration, the u-v coverage, the synthesized beam obtained for the proposed configuration.

  14. Investigations of two types of superconducting arrays

    International Nuclear Information System (INIS)

    Niu, M.

    1993-01-01

    This dissertation has two parts. Part one studies the anisotropy effect on homogeneous superconducting wire-networks, by using the Abrikosov approach. The networks assumed to have an infinite square lattice geometry. An anisotropy parameter R is defined to be the cross sectional area ratio of the vertical and horizontal strands. Many limiting behaviors of the order parameter distribution as R → ∞ are obtained. Many anisotropy-induced vortex configurational transitions are found at several Φ/Φ 0 values studied, and are investigated in detail. Part two studies the ground-state vortex configurations of the Josephson-coupled arrays of superconducting islands. The Ginzburg-Landau Josephson array model is used. With arrays of Penrose tiling geometry, the authors have found negative evidences against a proposed mechanism, and positive evidences for a new mechanism for generating commensurate states. But the mechanisms for the majority of the nontrivial commensurate states remain to be investigated. With arrays of infinite square lattice geometry, a temperature-induced vortex configurational transition at Φ/Φ 0 = 1/6 is found. The authors discover that the equilibrium vortex ground state of an infinite square-lattice array can occur in a unit cell of size other than q by q, or 2q by 2q, which has been widely accepted and commonly used so far

  15. All-Electrical Graphene DNA Sensor Array.

    Science.gov (United States)

    Abbott, Jeffrey; Ham, Donhee; Xu, Guangyu

    2017-01-01

    Electrical sensing of biomolecules has been an important pursuit due to the label-free operation and chip-scale construct such sensing modality can enable. In particular, electrical biomolecular sensors based on nanomaterials such as semiconductor nanowires, carbon nanotubes, and graphene have demonstrated high sensitivity, which in the case of nanowires and carbon nanotubes can surpass typical optical detection sensitivity. Among these nanomaterials, graphene is well suited for a practical candidate for implementing a large-scale array of biomolecular sensors, as its two-dimensional morphology is readily compatible with industry standard top-down fabrication techniques. In our recent work published in 2014 Nature Communications, we demonstrated these benefits by creating DNA sensor arrays from chemical vapor deposition (CVD) graphene. The present chapter, which is a review of this recent work, outlines procedures demonstrating the use of individual graphene sites of the array in dual roles--electrophoretic electrodes for site specific probe DNA immobilization and field effect transistor (FET) sensors for detection of target DNA hybridization. The 100 fM detection sensitivity achieved in 7 out of 8 graphene FET sensors in the array combined with the alternative use of the graphene channels as electrophoretic electrodes for probe deposition represent steps toward creating an all-electrical multiplexed DNA array.

  16. Performance measurements of hybrid PIN diode arrays

    International Nuclear Information System (INIS)

    Jernigan, J.G.; Arens, J.F.; Collins, T.; Herring, J.; Shapiro, S.L.; Wilburn, C.D.

    1990-05-01

    We report on the successful effort to develop hybrid PIN diode arrays and to demonstrate their potential as components of vertex detectors. Hybrid pixel arrays have been fabricated by the Hughes Aircraft Co. by bump bonding readout chips developed by Hughes to an array of PIN diodes manufactured by Micron Semiconductor Inc. These hybrid pixel arrays were constructed in two configurations. One array format having 10 x 64 pixels, each 120 μm square, and the other format having 256 x 256 pixels, each 30 μm square. In both cases, the thickness of the PIN diode layer is 300 μm. Measurements of detector performance show that excellent position resolution can be achieved by interpolation. By determining the centroid of the charge cloud which spreads charge into a number of neighboring pixels, a spatial resolution of a few microns has been attained. The noise has been measured to be about 300 electrons (rms) at room temperature, as expected from KTC and dark current considerations, yielding a signal-to-noise ratio of about 100 for minimum ionizing particles. 4 refs., 13 figs

  17. Detection of Coxiella burnetii in ticks by PCR and by PCR - Restriction Fragment Length Polymorphism (RFLP)

    International Nuclear Information System (INIS)

    2010-01-01

    Coxiella burnetii, as an obligata intracellular bacterium, is the etiologic agent of Q-fever. It is widely distributed in nature and is responsible for infection in various animals (cattle, sheep, goat) and humans. C. burnetii has been isolated from milk, ticks and human patients with acute and chronic Q fever. Ticks are the principal vectors and reservoirs of C. burnetii. Since over 40 species of ticks have been found to be infected with C. burnetii, ticks can serve as indicators of infection in nature. In this study, total of 2472 ticks (1446 female, 1021 male and 5 nymphs) were collected from 38 provinces of Turkey. The ticks were gathered into groups of 1 to 7 ticks as to the provinces, species and gender for DNA extraction. Following DNA extraction, the groups were examined for the presence of C. burtii by using the CB1and CB2. The ticks collected from the province of Denizli (56 in total) were gathered into 13 groups according to the species and gender. From these groups, 6 were positive for C. burnetii. The ticks collected from Ankara province, total of 160 ticks, were grouped into 53 as to their species and gender, only one group was found to be positive for C. burnetii. The specificities of PCR products were evaluated by restriction analysis. The positive PCR products were digested with the enzyme Taq1 and for bands in order of 118, 57, 43 and 39 bp's were appeared such as seen in the positive control DNA (C. burnetii Nine Mile RSA493)

  18. Exercising calf muscle T₂∗ changes correlate with pH, PCr recovery and maximum oxidative phosphorylation.

    Science.gov (United States)

    Schmid, Albrecht Ingo; Schewzow, Kiril; Fiedler, Georg Bernd; Goluch, Sigrun; Laistler, Elmar; Wolzt, Michael; Moser, Ewald; Meyerspeer, Martin

    2014-05-01

    Skeletal muscle metabolism is impaired in disorders like diabetes mellitus or peripheral vascular disease. The skeletal muscle echo planar imaging (EPI) signal (S(EPI) ) and its relation to energy metabolism are still debated. Localised ³¹P MRS and S(EPI) data from gastrocnemius medialis of 19 healthy subjects were combined in one scanning session to study direct relationships between phosphocreatine (PCr), pH kinetics and parameters of T₂∗ time courses. Dynamic spectroscopy (semi-LASER) and EPI were performed immediately before, during and after 5 min of plantar flexions. Data were acquired in a 7 T MR scanner equipped with a custom-built ergometer and a dedicated ³¹P/¹H radio frequency (RF) coil array. Using a form-fitted multi-channel ³¹P/¹H coil array resulted in high signal-to-noise ratio (SNR). PCr and pH in the gastrocnemius medialis muscle were quantified from each ³¹P spectrum, acquired every 6 s. During exercise, SEPI (t) was found to be a linear function of tissue pH(t) (cross-correlation r = -0.85 ± 0.07). Strong Pearson's correlations were observed between post exercise time-to-peak (TTP) of SEPI and (a) the time constant of PCr recovery τPCr recovery (r = 0.89, p energy metabolism were observed. In conclusion, a tight coupling between skeletal muscle metabolic activity and tissue T₂∗ signal weighting, probably induced by osmotically driven water shift, exists and can be measured non-invasively, using NMR at 7 T. © 2014 The Authors. NMR in Biomedicine published by John Wiley & Sons, Ltd.

  19. DNA extraction method for PCR in mycorrhizal fungi.

    Science.gov (United States)

    Manian, S; Sreenivasaprasad, S; Mills, P R

    2001-10-01

    To develop a simple and rapid DNA extraction protocol for PCR in mycorrhizal fungi. The protocol combines the application of rapid freezing and boiling cycles and passage of the extracts through DNA purification columns. PCR amplifiable DNA was obtained from a number of endo- and ecto-mycorrhizal fungi using minute quantities of spores and mycelium, respectively. DNA extracted following the method, was used to successfully amplify regions of interest from high as well as low copy number genes. The amplicons were suitable for further downstream applications such as sequencing and PCR-RFLPs. The protocol described is simple, short and facilitates rapid isolation of PCR amplifiable genomic DNA from a large number of fungal isolates in a single day. The method requires only minute quantities of starting material and is suitable for mycorrhizal fungi as well as a range of other fungi.

  20. Improvement of specific polymerase chain reaction (PCR) for the ...

    African Journals Online (AJOL)

    Improvement of specific polymerase chain reaction (PCR) for the identification of Mycoplasma capricolum subsp. capripneumoniae. H Ying, Z Nian-zhang, Z Ping, C Yue-feng, G Peng-cheng, Z Jian-jun, L Zhong-xin ...

  1. Chilean IPNV isolates: Robustness analysis of PCR detection

    Directory of Open Access Journals (Sweden)

    Esteban Jorquera

    2016-03-01

    Conclusions: Negative results must be interpreted carefully given the high genomic variability of IPNVs. Detection techniques (quantitative reverse transcription (qRT-PCR based on degenerate primers can be used to minimize the possibilities of false-negative detections.

  2. Simplex and triplex polymerase chain reaction (PCR) for ...

    African Journals Online (AJOL)

    Simplex and triplex polymerase chain reaction (PCR) for identification of three medically important Candida species. Nabil S Harmal, Alireza Khodavandi, Mohammed A Alshawsh, Farida Jamal, Zamberi Sekawi, Ng Kee Peng, Pei Pei Chong ...

  3. Typing of Y chromosome SNPs with multiplex PCR methods

    DEFF Research Database (Denmark)

    Sanchez Sanchez, Juan Jose; Børsting, Claus; Morling, Niels

    2005-01-01

    We describe a method for the simultaneous typing of Y-chromosome single nucleotide polymorphism (SNP) markers by means of multiplex polymerase chain reaction (PCR) strategies that allow the detection of 35 Y chromosome SNPs on 25 amplicons from 100 to 200 pg of chromosomal deoxyribonucleic acid...... (DNA). Multiplex PCR amplification of the DNA was performed with slight modifications of standard PCR conditions. Single-base extension (SBE) was performed using the SNaPshot kit containing fluorescently labeled ddNTPs. The extended primers were detected on an ABI 3100 sequencer. The most important...... factors for the creation of larger SNP typing PCR multiplexes include careful selection of primers for the primary amplification and the SBE reaction, use of DNA primers with homogenous composition, and balancing the primer concentrations for both the amplification and the SBE reactions....

  4. Comparison of conventional culture and real-time quantitative PCR ...

    African Journals Online (AJOL)

    2009-10-28

    HACCP) process in environments or during outbreak investigations this delay is a serious drawback. In recent years polymerase chain reaction (PCR) meth- ods have become the alternative to detect the presence of. Legionella in a ...

  5. Nanodroplet real-time PCR system with laser assisted heating

    Science.gov (United States)

    Kim, Hanyoup; Dixit, Sanhita; Green, Christopher J.; Faris, Gregory W.

    2011-01-01

    We report the successful application of low-power (~30 mW) laser radiation as an optical heating source for high-speed real-time polymerase chain reaction (PCR) amplification of DNA in nanoliter droplets dispersed in an oil phase. Light provides the heating, temperature measurement, and Taqman real-time readout in nanoliter droplets on a disposable plastic substrate. A selective heating scheme using an infrared laser appears ideal for driving PCR because it heats only the droplet, not the oil or plastic substrate, providing fast heating and completing the 40 cycles of PCR in 370 seconds. No microheaters or microfluidic circuitry were deposited on the substrate, and PCR was performed in one droplet without affecting neighboring droplets. The assay performance was quantitative and its amplification efficiency was comparable to that of a commercial instrument. PMID:19129891

  6. High-power, ultralow-mass solar arrays: FY-77 solar arrays technology readiness assessment report, volume 2

    Science.gov (United States)

    Costogue, E. N.; Young, L. E.; Brandhorst, H. W., Jr.

    1978-01-01

    Development efforts are reported in detail for: (1) a lightweight solar array system for solar electric propulsion; (2) a high efficiency thin silicon solar cell; (3) conceptual design of 200 W/kg solar arrays; (4) fluorocarbon encapsulation for silicon solar cell array; and (5) technology assessment of concentrator solar arrays.

  7. Comparison of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. in milk and meat samples

    DEFF Research Database (Denmark)

    Perelle, Sylvie; Dilasser, Françoise; Malorny, Burkhard

    2004-01-01

    , minced beef and raw milk, and 92 naturally-contaminated milk and meat samples. When using either PCR-ELISA or LC-PCR assays, only Salmonella strains were detected. PCR-ELISA and LC-PCR assays gave with pure Salmonella cultures the same detection limit level of 10(3) CFU/ml, which corresponds respectively...

  8. Development of RT-PCR and Nested PCR for Detecting Four Quarantine Plant Viruses Belonging to Nepovirus

    Directory of Open Access Journals (Sweden)

    Siwon Lee

    2013-09-01

    Full Text Available For quarantine purpose, we developed the RT- and nested PCR module of Tomato black ring virus (TBRV, Arabis mosaic virus (ArMV, Cherry leafroll virus (CLRV and Grapevine fanleaf virus (GFLV. The PCR modules, developed in this study make diagnosis more convenient and speedy because of same PCR condition. And also, the methods are more accurate because it can check whether the result is contamination or not using the mutation-positive control. We discard or return the 27 cases of Nepovirus infection seed by employing the module past 3 years. This study provides a rapid and useful method for detection of four quarantine plant viruses.

  9. Comparison among the Quantification of Bacterial Pathogens by qPCR, dPCR, and Cultural Methods

    Directory of Open Access Journals (Sweden)

    Matteo Ricchi

    2017-06-01

    Full Text Available The demand for rapid methods for the quantification of pathogens is increasing. Among these methods, those based on nucleic acids amplification (quantitative PCRs are the most widespread worldwide. Together with the qPCR, a new approach named digital PCR (dPCR, has rapidly gained importance. The aim of our study was to compare the results obtained using two different dPCR systems and one qPCR in the quantification of three different bacterial pathogens: Listeria monocytogenes, Francisella tularensis, and Mycobacterium avium subsp. paratuberculosis. For this purpose, three pre-existing qPCRs were used, while the same primers and probes, as well as PCR conditions, were transferred to two different dPCR systems: the QX200 (Bio-Rad and the Quant Studio 3D (Applied Biosystems. The limits of detection and limits of quantification for all pathogens, and all PCR approaches applied, were determined using genomic pure DNAs. The quantification of unknown decimal suspensions of the three bacteria obtained by the three different PCR approaches was compared through the Linear Regression and Bland and Altman analyses. Our results suggest that, both dPCRs are able to quantify the same amount of bacteria, while the comparison among dPCRs and qPCRs, showed both over and under-estimation of the bacteria present in the unknown suspensions. Our results showed qPCR over-estimated the amount of M. avium subsp. paratuberculosis and F. tularensis cells. On the contrary, qPCR, compared to QX200 dPCR, under-estimated the amount of L. monocytogenes cells. However, the maximum difference among PCRs approaches was <0.5 Log10, while cultural methods underestimated the number of bacteria by one to two Log10 for Francisella tularensis and Mycobacterium avium subsp. paratuberculosis. On the other hand, cultural and PCRs methods quantified the same amount of bacteria for L. monocytogenes, suggesting for this last pathogen, PCRs approaches can be considered as a valid alternative

  10. Implementation of High Resolution Whole Genome Array CGH in the Prenatal Clinical Setting: Advantages, Challenges, and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Paola Evangelidou

    2013-01-01

    Full Text Available Array Comparative Genomic Hybridization analysis is replacing postnatal chromosomal analysis in cases of intellectual disabilities, and it has been postulated that it might also become the first-tier test in prenatal diagnosis. In this study, array CGH was applied in 64 prenatal samples with whole genome oligonucleotide arrays (BlueGnome, Ltd. on DNA extracted from chorionic villi, amniotic fluid, foetal blood, and skin samples. Results were confirmed with Fluorescence In Situ Hybridization or Real-Time PCR. Fifty-three cases had normal karyotype and abnormal ultrasound findings, and seven samples had balanced rearrangements, five of which also had ultrasound findings. The value of array CGH in the characterization of previously known aberrations in five samples is also presented. Seventeen out of 64 samples carried copy number alterations giving a detection rate of 26.5%. Ten of these represent benign or variables of unknown significance, giving a diagnostic capacity of the method to be 10.9%. If karyotype is performed the additional diagnostic capacity of the method is 5.1% (3/59. This study indicates the ability of array CGH to identify chromosomal abnormalities which cannot be detected during routine prenatal cytogenetic analysis, therefore increasing the overall detection rate. In addition a thorough review of the literature is presented.

  11. Development of Candida-Specific Real-Time PCR Assays for the Detection and Identification of Eight Medically Important Candida Species.

    Science.gov (United States)

    Zhang, Jing; Hung, Guo-Chiuan; Nagamine, Kenjiro; Li, Bingjie; Tsai, Shien; Lo, Shyh-Ching

    2016-01-01

    Culture-based identification methods have been the gold standard for the diagnosis of fungal infection. Currently, molecular technologies such as real-time PCR assays with short turnaround time can provide desirable alternatives for the rapid detection of Candida microbes. However, most of the published PCR primer sets are not Candida specific and likely to amplify DNA from common environmental contaminants, such as Aspergillus microbes. In this study, we designed pan-Candida primer sets based on the ribosomal DNA-coding regions conserved within Candida but distinct from those of Aspergillus and Penicillium. We demonstrate that the final two selected pan-Candida primer sets would not amplify Aspergillus DNA and could be used to differentiate eight medically important Candida pathogens in real-time PCR assays based on their melting profiles, with a sensitivity of detection as low as 10 fg of Candida genomic DNA. Moreover, we further evaluated and selected species-specific primer sets covering Candida albicans, Candida glabrata, Candida tropicalis, and Candida dubliniensis and show that they had high sensitivity and specificity. These real-time PCR primer sets could potentially be assembled into a single PCR array for the rapid detection of Candida species in various clinical settings, such as corneal transplantation.

  12. Detection of alcohol-tolerant hiochi bacteria by PCR.

    OpenAIRE

    Nakagawa, T; Shimada, M; Mukai, H; Asada, K; Kato, I; Fujino, K; Sato, T

    1994-01-01

    We report a sensitive and rapid method for detection of hiochi bacteria by PCR. This method involves the electrophoresis of amplified DNA. Nucleotide sequences of the spacer region between 16S and 23S rRNA genes of 11 Lactobacillus strains were identified by analysis of PCR products. Five primers were designed by analysis of similarities among these sequences. A single cell of Lactobacillus casei subsp. casei could be detected when purified genomic DNA was used as the template. When various c...

  13. Identification of Phytophthora fragariae var. rubi by PCR.

    Science.gov (United States)

    Schlenzig, Alexandra

    2009-01-01

    The following chapter describes a PCR method for the identification of the raspberry root rot pathogen Phytophthora fragariae var. rubi. Furthermore, a nested PCR suitable for the detection of the pathogen in infected raspberry roots and validated against the "Duncan bait test" (EPPO Bull 35:87-91, 2005) is explained. Protocols for different DNA extraction methods are given which can be transferred to other fungal pathogens.

  14. MPprimer: a program for reliable multiplex PCR primer design

    Directory of Open Access Journals (Sweden)

    Wang Xiaolei

    2010-03-01

    Full Text Available Abstract Background Multiplex PCR, defined as the simultaneous amplification of multiple regions of a DNA template or multiple DNA templates using more than one primer set (comprising a forward primer and a reverse primer in one tube, has been widely used in diagnostic applications of clinical and environmental microbiology studies. However, primer design for multiplex PCR is still a challenging problem and several factors need to be considered. These problems include mis-priming due to nonspecific binding to non-target DNA templates, primer dimerization, and the inability to separate and purify DNA amplicons with similar electrophoretic mobility. Results A program named MPprimer was developed to help users for reliable multiplex PCR primer design. It employs the widely used primer design program Primer3 and the primer specificity evaluation program MFEprimer to design and evaluate the candidate primers based on genomic or transcript DNA database, followed by careful examination to avoid primer dimerization. The graph-expanding algorithm derived from the greedy algorithm was used to determine the optimal primer set combinations (PSCs for multiplex PCR assay. In addition, MPprimer provides a virtual electrophotogram to help users choose the best PSC. The experimental validation from 2× to 5× plex PCR demonstrates the reliability of MPprimer. As another example, MPprimer is able to design the multiplex PCR primers for DMD (dystrophin gene which caused Duchenne Muscular Dystrophy, which has 79 exons, for 20×, 20×, 20×, 14×, and 5× plex PCR reactions in five tubes to detect underlying exon deletions. Conclusions MPprimer is a valuable tool for designing specific, non-dimerizing primer set combinations with constrained amplicons size for multiplex PCR assays.

  15. Terabyte IDE RAID-5 Disk Arrays

    Energy Technology Data Exchange (ETDEWEB)

    David A. Sanders et al.

    2003-09-30

    High energy physics experiments are currently recording large amounts of data and in a few years will be recording prodigious quantities of data. New methods must be developed to handle this data and make analysis at universities possible. We examine some techniques that exploit recent developments in commodity hardware. We report on tests of redundant arrays of integrated drive electronics (IDE) disk drives for use in offline high energy physics data analysis. IDE redundant array of inexpensive disks (RAID) prices now are less than the cost per terabyte of million-dollar tape robots! The arrays can be scaled to sizes affordable to institutions without robots and used when fast random access at low cost is important.

  16. Microneedle arrays for biosensing and drug delivery

    Science.gov (United States)

    Wang, Joseph; Windmiller, Joshua Ray; Narayan, Roger; Miller, Philip; Polsky, Ronen; Edwards, Thayne L.

    2017-08-22

    Methods, structures, and systems are disclosed for biosensing and drug delivery techniques. In one aspect, a^ device for detecting an analyte and/or releasing a biochemical into a biological fluid can include an array of hollowed needles, in which each needle includes a protruded needle structure including an exterior wall forming a hollow interior and an opening at a terminal end of the protruded needle structure that exposes the hollow interior, and a probe inside the exterior wall to interact with one or more chemical or biological substances that come in contact with the probe via the opening to produce a probe sensing signal, and an array of wires that are coupled to probes of the array of hollowed needles, respectively, each wire being electrically conductive to transmit the probe sensing signal produced by a respective probe.

  17. Microneedle arrays for biosensing and drug delivery

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Joseph; Windmiller, Joshua Ray; Narayan, Roger; Miller, Philip

    2017-08-29

    Methods, structures, and systems are disclosed for biosensing and drug delivery techniques. In one aspect, a device for detecting an analyte and/or releasing a biochemical into a biological fluid can include an array of hollowed needles, in which each needle includes a protruded needle structure including an exterior wall forming a hollow interior and an opening at a terminal end of the protruded needle structure that exposes the hollow interior, and a probe inside the exterior wall to interact with one or more chemical or biological substances that come in contact with the probe via the opening to produce a probe sensing signal, and an array of wires that are coupled to probes of the array of hollowed needles, respectively, each wire being electrically conductive to transmit the probe sensing signal produced by a respective probe.

  18. Big data challenges for large radio arrays

    Science.gov (United States)

    Jones, D. L.; Wagstaff, K.; Thompson, D. R.; D'Addario, L.; Navarro, R.; Mattmann, C.; Majid, W.; Lazio, J.; Preston, J.; Rebbapragada, U.

    2012-03-01

    Future large radio astronomy arrays, particularly the Square Kilometre Array (SKA), will be able to generate data at rates far higher than can be analyzed or stored affordably with current practices. This is, by definition, a "big data" problem, and requires an end-to-end solution if future radio arrays are to reach their full scientific potential. Similar data processing, transport, storage, and management challenges face next-generation facilities in many other fields. The Jet Propulsion Laboratory is developing technologies to address big data issues, with an emphasis in three areas: 1) Lower-power digital processing architectures to make highvolume data generation operationally affordable, 2) Date-adaptive machine learning algorithms for real-time analysis (or "data triage") of large data volumes, and 3) Scalable data archive systems that allow efficient data mining and remote user code to run locally where the data are stored.

  19. Systems and methods of manufacturing microchannel arrays

    Energy Technology Data Exchange (ETDEWEB)

    Paul, Brian K.; Brannon, Samuel T.

    2018-03-20

    The present application relates to apparatus and methods of reducing the cost of microchannel array production and operation. In a representative embodiment, a microchannel array can comprise a first lamina having one or more flanges and a plurality of elongated bosses. The one or more flanges can extend along a perimeter of the first lamina, the plurality of elongated bosses can at least partially define a plurality of first flow paths, and the first lamina can define at least one opening. The microchannel array can also comprise a second lamina having a plurality of second flow paths, and can define at least one opening. The second lamina can be disposed above the first lamina such that the second lamina encloses the first flow paths of the first lamina and the at least one opening of the first lamina is coaxial with the at least one opening of the second lamina.

  20. Stochastic synaptic plasticity with memristor crossbar arrays

    KAUST Repository

    Naous, Rawan

    2016-11-01

    Memristive devices have been shown to exhibit slow and stochastic resistive switching behavior under low-voltage, low-current operating conditions. Here we explore such mechanisms to emulate stochastic plasticity in memristor crossbar synapse arrays. Interfaced with integrate-and-fire spiking neurons, the memristive synapse arrays are capable of implementing stochastic forms of spike-timing dependent plasticity which parallel mean-rate models of stochastic learning with binary synapses. We present theory and experiments with spike-based stochastic learning in memristor crossbar arrays, including simplified modeling as well as detailed physical simulation of memristor stochastic resistive switching characteristics due to voltage and current induced filament formation and collapse. © 2016 IEEE.