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Sample records for antibody test development

  1. Development of a blocking latex agglutination test for the detection of antibodies to chicken anemia virus.

    Science.gov (United States)

    Trinh, Dai Quang; Ogawa, Haruko; Bui, Vuong Nghia; Nguyen, Tham Thi Hong; Gronsang, Dulyatad; Baatartsogt, Tugsbaatar; Kizito, Mugimba Kahoza; AboElkhair, Mohammed; Yamaguchi, Shigeo; Nguyen, Viet Khong; Imai, Kunitoshi

    2015-09-01

    A blocking latex agglutination test (b-LAT) developed in this study was evaluated for the detection of antibodies against chicken anemia virus (CAV) in chickens. Polystyrene latex beads were coupled with a neutralizing monoclonal antibody (mAb) to CAV (mAb-beads). When mAb-beads were mixed with antigens prepared from the lysate of MDCC-MSB1 cells infected with CAV, agglutination occurred. A short pre-incubation of CAV antigens with CAV-specific antiserum inhibited the agglutination of mAb-beads. The test results were obtained within 5min. The specificity of b-LAT was evaluated using sera from specific pathogen-free chickens and sera containing antibodies to avian influenza virus, Newcastle disease virus, infectious bursal disease virus, and Marek's disease virus; nonspecific agglutination and cross-reactivity with antibodies to unrelated viruses were not observed. The examination of 94 serum samples collected from commercial breeder chickens of various ages (17-63 weeks) revealed good agreement (93.6%, Kappa value=0.82) between b-LAT and a virus neutralization test, known to be most sensitive and specific in the detection of antibodies to CAV. These results indicate that b-LAT, a simple and rapid test, is a useful and reliable tool in CAV serology. PMID:25952731

  2. HIV Antibody Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? HIV Antibody and HIV Antigen (p24) Share this page: Was this page helpful? Also known as: HIV Screening Tests; AIDS Test; AIDS Screen; HIV Serology; ...

  3. The development of a radioallergosorbent test (RAST) for murine IgE antibodies

    International Nuclear Information System (INIS)

    A purified monoclonal IgE preparation, isolated from the ascitic fluid of mice bearing a hybridoma secreting IgE with specificity to ovalbumin, was used for the production of goat anti-murine IgE (GAME) antiserum, which was then rendered monospecific for the epsilon chain. Another monoclonal hybridoma IgE preparation with specificity for the 2,4-dinitrophenyl group was isolated from ascitic fluid in a relatively pure state by affinity chromatography and used in the form of an immunosorbent to isolate antibodies from the monospecific goat serum. The GAME of antibodies were 125I-labeled and used to develop a radioallergosorbent test (RAST) for the quantitation of murine IgE antibodies specifically adsorbed onto antigen-coupled paper discs. The RAST was specific for antibodies of the IgE class only and was as sensitive as and more accurate than PCA assay. RAST results on sera of mice treated with tolerogenic conjugates indicated a reduction in the affinity and concentration of the IgE antibody populations on suppression of the IgE response. The effect of interference by non-IgE antibody populations on the RAST curves has been discussed. (Auth.)

  4. Anti-insulin antibody test

    Science.gov (United States)

    Insulin antibodies - serum; Insulin Ab test ... Normally, there are no antibodies against insulin in your blood. Normal value ranges may vary slightly among different laboratories. Some labs use different measurements or ...

  5. Epstein-Barr virus antibody test

    Science.gov (United States)

    EBV antibody test; EBV serology ... a lab, where a lab specialist looks for antibodies to the Epstein-Barr virus. In the first stages of an illness, little antibody may be detected. For this reason, the test ...

  6. Development and evaluation of two simple, rapid immunochromatographic tests for the detection of Yersinia pestis antibodies in humans and reservoirs.

    Directory of Open Access Journals (Sweden)

    Minoarisoa Rajerison

    Full Text Available BACKGROUND: Tools for plague diagnosis and surveillance are not always available and affordable in most of the countries affected by the disease. Yersinia pestis isolation for confirmation is time-consuming and difficult to perform under field conditions. Serologic tests like ELISA require specific equipments not always available in developing countries. In addition to the existing rapid test for antigen detection, a rapid serodiagnostic assay may be useful for plague control. METHODS/PRINCIPAL FINDINGS: We developed two rapid immunochromatography-based tests for the detection of antibodies directed against F1 antigen of Y. pestis. The first test, SIgT, which detects total Ig (IgT anti-F1 in several species (S (human and reservoirs, was developed in order to have for the field use an alternative method to ELISA. The performance of the SIgT test was evaluated with samples from humans and animals for which ELISA was used to determine the presumptive diagnosis of plague. SIgT test detected anti-F1 Ig antibodies in humans with a sensitivity of 84.6% (95% CI: 0.76-0.94 and a specificity of 98% (95% CI: 0.96-1. In evaluation of samples from rodents and other small mammals, the SlgT test had a sensitivity of 87.8% (95% CI: 0.80-0.94 and a specificity of 90.3% (95% CI: 0.86-0.93. Improved performance was obtained with samples from dogs, a sentinel animal, with a sensitivity of 93% (95% CI: 0.82-1 and a specificity of 98% (95% CI: 0.95-1.01. The second test, HIgM, which detects human (H IgM anti-F1, was developed in order to have another method for plague diagnosis. Its sensitivity was 83% (95% CI: 0.75-0.90 and its specificity about 100%. CONCLUSION/SIGNIFICANCE: The SIgT test is of importance for surveillance because it can detect Ig antibodies in a range of reservoir species. The HIgM test could facilitate the diagnosis of plague during outbreaks, particularly when only a single serum sample is available.

  7. Development of a immunochromatographic test with avidin-biotin for the detection of antibodies against antigen e of hepatitis B in human plasma

    International Nuclear Information System (INIS)

    The disappearance of antigen e of hepatitis B in the presence of the plasmatic antibodies against antigen e may indicate a satisfactory therapeutic response in patients with chronic hepatitis B. The immuno-chromatographic test carried out in the diagnosis of diseases use different antibody combinations and may employ the avidin or streptavidin-biotin technology to develop a rapid immuno-chromatographic test for the detection of antibodies anti-antigen e in the plasma. They were detected in the laboratory by means of two fast immuno-chromatographic tests when using in one of them the avidin-biotin technology. These tests are carried out with a one-step competitive inhibition format and amplified or not with avidin-biotin. Monoclonal antibodies against antigen e obtained by cellular hybridization were used. Forty-six plasmatic samples classified as positive and negative to the anti-antigen antibodies were evaluated with a reference immunochromatographic test Advanced QualityTM. The possible expiry time of the biological reagents forming part of these tests were studied with accelerated thermal-stability experiments. The possible interference in the plasma of some of the biochemical compounds used in these trials was analyzed. Four murine monoclonal antibodies anti-antigen e were obtained and only one of them was used in these immunochromatographic tests with an anti-antigen polyclonal antibody conjugated with gold. Both tests and their stable biological reagents discriminated the positive and negative samples to the antibodies anti-antigen e, as well as the commercial test. There was no interference in the biochemical compounds studied in these tests. Both immuno-chromatographic tests made in the laboratory are useful to detect antibodies anti-antigen e in the plasma. The avidin-biotin increased the analytical sensitivity of this type of fast immuno-chromatographic test without altering its performance features. (Author)

  8. TSOL18 Vaccine Antigen of Taenia solium: Development of Monoclonal Antibodies and Field Testing of the Vaccine in Cameroon

    Directory of Open Access Journals (Sweden)

    Assana, E.

    2010-01-01

    lack of knowledge of the taeniasis-cysticercosis complex and the absence of a pig pen in the household were associated with pig cysticercosis. Chapter 3 reports the investigations that were undertaken to characterise whether the principal antibody specificities raised by TSOL18 in pigs were against linear or conformational determinants. TSOL18 was expressed in two truncated forms representing either the amino terminal portion or the carboxy terminal portion, with the two truncations overlapping in sequence by 25 amino acids. The original protein (designated TSOL18N— and the two truncations (TSOL18N—-1 and TSOL18N—-2 were used in inhibition ELISA to determine their ability to inhibit the binding of protective pig antibodies to TSOL18. TSOL18N— was shown to be capable of completely inhibiting the binding of pig anti-TSOL18N— antibodies to TSOL18N— in ELISA. However, neither TSOL18N—-1 nor TSOL18N—-2, either alone or combined, was capable of inhibiting any detectable amount of reactivity of pig anti-TSOL18N— antibodies with TSOL18N—. It is concluded that the dominant antibody specificities, and likely the host-protective specificities, of TSOL18 are conformational epitopes. Chapter 4 describes the development of an antibody detection test for the specific diagnosis of porcine cysticercosis. A fraction with a major band of 14 kDa was obtained from crude cyst fluid (CF of T. solium cysticerci by 2-step chromatography. A first fraction isolated by gel filtration was purified using an anion exchange column on High Performance Liquid Chromatography (HPLC. Evaluation of the analytic sensitivity of this fraction (F3 was carried out in an antibody detection enzyme-linked immunosorbent assay (Ab-ELISA-F3 using serum samples from pigs experimentally infected with different doses of T. solium eggs. The cross-reactivity of F3 was evaluated with serum samples from pigs that were naturally or experimentally infected with Taenia hydatigena, Taenia saginata asiatica

  9. Metrics for antibody therapeutics development

    OpenAIRE

    Reichert, Janice M

    2010-01-01

    A wide variety of full-size monoclonal antibodies (mAbs) and therapeutics derived from alternative antibody formats can be produced through genetic and biological engineering techniques. These molecules are now filling the preclinical and clinical pipelines of every major pharmaceutical company and many biotechnology firms. Metrics for the development of antibody therapeutics, including averages for the number of candidates entering clinical study and development phase lengths for mAbs approv...

  10. Development and evaluation of a rapid latex agglutination test using a monoclonal antibody to identify Candida dubliniensis colonies.

    Science.gov (United States)

    Marot-Leblond, Agnes; Beucher, Bertrand; David, Sandrine; Nail-Billaud, Sandrine; Robert, Raymond

    2006-01-01

    Cell components of the dimorphic pathogenic fungus Candida dubliniensis were used to prepare monoclonal antibodies (MAbs). One MAb, designated 12F7-F2, was shown by indirect immunofluorescence to be specific for a surface antigen of Candida dubliniensis yeast cells. No reactivity was observed with other fungal genera or with other Candida species, including Candida albicans, that share many phenotypic features with C. dubliniensis. The use of different chemical and physical treatments for cell component extraction suggested that the specific epitope probably resides on a protein moiety absent from C. albicans. However, we failed to identify the target protein by Western blotting, owing to its sensitivity to heat and sodium dodecyl sulfate. MAb 12F7-F2 was further used to develop a commercial latex agglutination test to identify C. dubliniensis colonies (Bichro-dubli Fumouze test; Fumouze Diagnostics). The test was validated on yeast strains previously identified by PCR and on fresh clinical isolates; these included 46 C. dubliniensis isolates, 45 C. albicans isolates, and other yeast species. The test had 100% sensitivity and specificity for C. dubliniensis isolated on Sabouraud dextrose, CHROMagar Candida, and CandiSelect media and 97.8% sensitivity for C. dubliniensis grown on Candida ID medium. The test is rapid (5 min) and easy to use and may be recommended for routine use in clinical microbiology laboratories and for epidemiological investigations. PMID:16390961

  11. Heparin-Induced Thrombocytopenia Antibody Test

    Science.gov (United States)

    ... Thrombocytopenia Platelet Factor 4 Antibody Related tests: Complete Blood Count , Platelet Count , Serotonin Release Assay, Heparin-induced Platelet Aggregation All content on Lab Tests Online has been ...

  12. Metrics for antibody therapeutics development.

    Science.gov (United States)

    Reichert, Janice M

    2010-01-01

    A wide variety of full-size monoclonal antibodies (mAbs) and therapeutics derived from alternative antibody formats can be produced through genetic and biological engineering techniques. These molecules are now filling the preclinical and clinical pipelines of every major pharmaceutical company and many biotechnology firms. Metrics for the development of antibody therapeutics, including averages for the number of candidates entering clinical study and development phase lengths for mAbs approved in the United States, were derived from analysis of a dataset of over 600 therapeutic mAbs that entered clinical study sponsored, at least in part, by commercial firms. The results presented provide an overview of the field and context for the evaluation of on-going and prospective mAb development programs. The expansion of therapeutic antibody use through supplemental marketing approvals and the increase in the study of therapeutics derived from alternative antibody formats are discussed. PMID:20930555

  13. Development of Monoclonal Antibody and Diagnostic Test for Middle East Respiratory Syndrome Coronavirus Using Cell-Free Synthesized Nucleocapsid Antigen

    Science.gov (United States)

    Yamaoka, Yutaro; Matsuyama, Shutoku; Fukushi, Shuetsu; Matsunaga, Satoko; Matsushima, Yuki; Kuroyama, Hiroyuki; Kimura, Hirokazu; Takeda, Makoto; Chimuro, Tomoyuki; Ryo, Akihide

    2016-01-01

    Protein nativity is one of the most critical factors for the quality of antigens used as immunogens and the reactivities of the resultant antibodies. The preparation and purification of native viral antigens in conventional cell-based protein expression systems are often accompanied by technical hardships. These challenges are attributable mainly to protein aggregation and insolubility during expression and purification, as well as to very low expression levels associated with the toxicity of some viral proteins. Here, we describe a novel approach for the production of monoclonal antibodies (mAbs) against nucleocapsid protein (NP) of the Middle East respiratory syndrome coronavirus (MERS-CoV). Using a wheat germ cell-free protein synthesis system, we successfully prepared large amounts of MERS-CoV NP antigen in a state that was highly soluble and intact for immunization. Following mouse immunization and hybridoma generation, we selected seven hybridoma clones that produced mAbs with exclusive reactivity against MERS-CoV NP. Epitope mapping and subsequent bioinformatic analysis revealed that these mAbs recognized epitopes located within relatively highly conserved regions of the MERS-CoV amino-acid sequence. Consistently, the mAbs exhibited no obvious cross-reactivity with NPs derived from other related viruses, including SARS coronavirus. After determining the optimal combinations of these mAbs, we developed an enzyme-linked immunosorbent assay and a rapid immunochromatographic antigen detection test that can be reliably used for laboratory diagnosis of MERS-CoV. Thus, this study provides strong evidence that the wheat germ cell-free system is useful for the production of diagnostic mAbs against emerging pathogens. PMID:27148198

  14. Development of immunochromatographic strip test using fluorescent, micellar silica nanosensors for rapid detection of B. abortus antibodies in milk samples.

    Science.gov (United States)

    Vyas, Swati S; Jadhav, Sushma V; Majee, Sharmila B; Shastri, Jayanthi S; Patravale, Vandana B

    2015-08-15

    Presence of bacteria such as Brucella spp. in dairy products is an immense risk to public health. Point of care immunoassays are rapid in that they can quickly screen various samples in a relatively short amount of time, are sensitive, specific and offer a great advantage in accurate and fast diagnosis of infectious diseases. We have fabricated a point of care rapid diagnostic assay that employs fluorescent, micellar silica nanosensors capable of specifically detecting Brucella IgG antibodies in milk samples of afflicted animals. Currently, point of care detection assays are not commercially available for field testing of farm animals using milk samples. The nanosensing allows precise detection of antibodies with low sample volumes (50 μl). We demonstrate recognition of B. abortus antibodies through capture by fluorescent silica nanosensors using spiked and raw milk samples validated by ELISA and PCR. The test results are accurate and repeatable with high sensitivity and specificity, and a short assay time of 10 min for antigenic recognition and do not require any sample processing procedures such as isolation and separation. Additionally, well defined antigenic components and surface biomarkers of various disease causing microbes can be broadly incorporated within the purview of this technology for accurate and rapid detection of suspected bovine pathological conditions, and can largely enable rapid field testing that can be implemented in farms and food industry. PMID:25829223

  15. Quality control of antibodies for assay development.

    Science.gov (United States)

    Schumacher, Sarah; Seitz, Harald

    2016-09-25

    Antibodies are used as powerful tools in basic research, for example, in biomarker identification, and in various forms for diagnostics, for example, identification of allergies or autoimmune diseases. Due to their robustness and ease of handling, immunoassays are favourite methods for investigation of various biological or medical questions. Nevertheless in many cases, additional analyses such as mass spectrometry are used to validate or confirm the results of immunoassays. To minimize the workload and to increase confidence in immunoassays, there are urgent needs for antibodies which are both highly specific and well validated. Unfortunately many commercially available antibodies are neither well characterized nor fully tested for cross-reactivities. Adequate quality control and validation of an antibody is time-consuming and can be frustrating. Such validation needs to be performed for every assay/application. However, where an antibody validation is successful, a highly specific and stable reagent will be on hand. This article describes the validation processes of antibodies, including some often neglected factors, as well as unspecific binding to other sample compounds in a multiparameter diagnostic assay. The validation consists of different immunological methods, with important assay controls, and is performed in relation to the development of a diagnostic test. PMID:26873787

  16. A protein-conjugate approach to develop a monoclonal antibody-based antigen detection test for the diagnosis of human brucellosis.

    Directory of Open Access Journals (Sweden)

    Kailash P Patra

    2014-06-01

    Full Text Available Human brucellosis is most commonly diagnosed by serology based on agglutination of fixed Brucella abortus as antigen. Nucleic acid amplification techniques have not proven capable of reproducibly and sensitively demonstrating the presence of Brucella DNA in clinical specimens. We sought to optimize a monoclonal antibody-based assay to detect Brucella melitensis lipopolysaccharide in blood by conjugating B. melitensis LPS to keyhole limpet hemocyanin, an immunogenic protein carrier to maximize IgG affinity of monoclonal antibodies. A panel of specific of monoclonal antibodies was obtained that recognized both B. melitensis and B. abortus lipopolysaccharide epitopes. An antigen capture assay was developed that detected B. melitensis in the blood of experimentally infected mice and, in a pilot study, in naturally infected Peruvian subjects. As a proof of principle, a majority (7/10 of the patients with positive blood cultures had B. melitensis lipopolysaccharide detected in the initial blood specimen obtained. One of 10 patients with relapsed brucellosis and negative blood culture had a positive serum antigen test. No seronegative/blood culture negative patients had a positive serum antigen test. Analysis of the pair of monoclonal antibodies (2D1, 2E8 used in the capture ELISA for potential cross-reactivity in the detection of lipopolysaccharides of E. coli O157:H7 and Yersinia enterocolitica O9 showed specificity for Brucella lipopolysaccharide. This new approach to develop antigen-detection monoclonal antibodies against a T cell-independent polysaccharide antigen based on immunogenic protein conjugation may lead to the production of improved rapid point-of-care-deployable assays for the diagnosis of brucellosis and other infectious diseases.

  17. 21 CFR 866.3290 - Gonococcal antibody test (GAT).

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Gonococcal antibody test (GAT). 866.3290 Section... antibody test (GAT). (a) Identification. A gonococcal antibody test (GAT) is an in vitro device that..., indirect fluorescent antibody, or radioimmunoassay, antibodies to Neisseria gonorrhoeae in sera...

  18. Utility of feline coronavirus antibody tests.

    Science.gov (United States)

    Addie, Diane D; le Poder, Sophie; Burr, Paul; Decaro, Nicola; Graham, Elizabeth; Hofmann-Lehmann, Regina; Jarrett, Oswald; McDonald, Michael; Meli, Marina L

    2015-02-01

    Eight different tests for antibodies to feline coronavirus (FCoV) were evaluated for attributes that are important in situations in veterinary practice. We compared four indirect immunofluorescent antibody tests (IFAT), one enzyme-linked immunosorbent assay (ELISA) (FCoV Immunocomb; Biogal) and three rapid immunochromatographic (RIM) tests against a panel of samples designated by consensus as positive or negative. Specificity was 100% for all but the two IFATs based on transmissible gastroenteritis virus (TGEV), at 83.3% and 97.5%. The IFAT and ELISA tests were best for obtaining an antibody titre and for working in the presence of virus. The RIM tests were the best for obtaining a result quickly (10-15 mins); of these, the Speed F-Corona was the most sensitive, at 92.4%, followed by FASTest feline infectious peritonitis (FIP; 84.6%) and Anigen Rapid FCoV antibody test (64.1%). Sensitivity was 100% for the ELISA, one FCoV IFAT and one TGEV IFAT; and 98.2% for a second TGEV IFA and 96.1% for a second FCoV IFAT. All tests worked with effusions, even when only blood products were stipulated in the instruction manual. The ELISA and Anigen RIM tests were best for small quantities of sample. The most appropriate FCoV antibody test to use depends on the reason for testing: in excluding a diagnosis of FIP, sensitivity, specificity, small sample quantity, rapidity and ability to work in the presence of virus all matter. For FCoV screening, speed and sensitivity are important, and for FCoV elimination antibody titre is essential. PMID:24966245

  19. Development of antibody against sulfamethazine

    International Nuclear Information System (INIS)

    Sulfamethazine (SMT) is widely used to treat bacterial and protozoan infections in food animals. So its residue has been detected in various food products, and in Europe, the tolerance level for sulfonamides in meat and milk is 100 ng/g. To ensure that residues in animal food products do not exceed this limit, a simple, sensitive, and rapid method to determinate their residues in animal tissues is needed. In this paper the development of polyclonal or monoclonal antibodies against sulfamethazine (SMT) and a simplified method to identify residual sulfamethazine by radio immunoassay (RIA) is presented. Polyclonal antibodies (PcAbs) against sulfamethazine (SMT) were obtained by immunizing rabbits with SMT-conjugated bovine serum albumin (BSA). The association constants (Ka) of the PcAbs were higher than 108 and the cross-reactivities with Sulfadiazine(SD), Sulfaquinoxaline(SQX) which were structurally related compounds were lower than 0.05%(RIA). Simultaneous, six strains of hybridoma cell were prepared which can secrete monoclonal antibodies (McAbs) against SMT . The Ka of the McAbs against SMT were higher than 107 and the cross-reactivities with SD, SQX were lower than 0.1%(RIA). (authors)

  20. Serum Treponema IgM Antibody Test for Syphilis Diagnosis

    Institute of Scientific and Technical Information of China (English)

    郑占才; 张荣富; 溪茜

    2003-01-01

    Objective: To evaluate the clinical utility of testing serum anti-treponema pallidum IgM antibody in the diagnosis of syphilis patients. Methods: Seventy-two cases of syphilis were tested for specific IgM antibody with ELISA, and the results were compared with RPR and TPPA.Results: The sensitivity of IgM antibody was 73.3 %(11/15) in primary syphilis, 88.9% (16/18) in sec-ondary syphilis, and there was no significant differ-ence between these values (x2=1.6363, P>0.10). The sensitivity of IgM antibody in diagnosing latent syphi-lis was only 26.1% (6/23), much lower than the detec-tion rate in symptomatic earlv svDhilis (x2=17.6189. P<0.005). RPR and TPPA were both 100% sensitive in latent and early symptomatic syphilis. Two were posi,five for IgM in the 16 cases who had received regular treatments 2 to 24 months before enrolled.Conclusions: Specific IgM antibody detection doees not appear superior to RPR and TPPA in diagnosing primary syphilis. The diagnosis of latent syphilis should mainly rely on RPR and TPPA, since there are low titers of IgM antibody at that stage. IgM antibody testing alone should not be recommended for monitor-ing syphilis development or treatment efficacy. Fur-ther studies should be concerned.

  1. Development and application of an indirect ELISA test for the detection of antibodies to Mycoplasma crocodyli infection in crocodiles (Crocodylus niloticus).

    Science.gov (United States)

    Dawo, Fufa; Mohan, Krishna

    2007-01-31

    Non-availability of a standardized rapid serodiagnostic test for quick and accurate diagnosis of Mycoplasma crocodyli (M. crocodyli) infection in crocodiles was the underlining reason for conducting the present study. An indirect enzyme-linked immunosorbent assay (iELISA) for the detection of antibodies (Ab) to M. crocodyli infection in crocodile sera was developed using sonicated antigen (Ag) and anti-crocodile conjugate. The iELISA test was optimised with different reagents and at different steps. A cut-off value of percent positive greater than or equal to 53.47% resulted in an estimated sensitivity and specificity of 85.67 and 100%, respectively. The developed iELISA could be used for detection of Abs to M. crocodyli infection in crocodiles and may enable to understand the transmission of the disease. PMID:17014973

  2. Development and Evaluation of a Rapid Latex Agglutination Test Using a Monoclonal Antibody To Identify Candida dubliniensis Colonies

    OpenAIRE

    Marot-Leblond, Agnes; Beucher, Bertrand; David, Sandrine; Nail-Billaud, Sandrine; Robert, Raymond

    2006-01-01

    Cell components of the dimorphic pathogenic fungus Candida dubliniensis were used to prepare monoclonal antibodies (MAbs). One MAb, designated 12F7-F2, was shown by indirect immunofluorescence to be specific for a surface antigen of Candida dubliniensis yeast cells. No reactivity was observed with other fungal genera or with other Candida species, including Candida albicans, that share many phenotypic features with C. dubliniensis. The use of different chemical and physical treatments for cel...

  3. 21 CFR 866.5110 - Antiparietal antibody immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antiparietal antibody immunological test system....5110 Antiparietal antibody immunological test system. (a) Identification. An antiparietal antibody... the specific antibody for gastric parietal cells in serum and other body fluids. Gastric...

  4. 21 CFR 866.5100 - Antinuclear antibody immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antinuclear antibody immunological test system....5100 Antinuclear antibody immunological test system. (a) Identification. An antinuclear antibody... the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular...

  5. 21 CFR 866.5090 - Antimitochondrial antibody immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antimitochondrial antibody immunological test... Systems § 866.5090 Antimitochondrial antibody immunological test system. (a) Identification. An antimitochondrial antibody immunological test system is a device that consists of the reagents used to measure...

  6. 21 CFR 660.54 - Potency tests, specificity tests, tests for heterospecific antibodies, and additional tests for...

    Science.gov (United States)

    2010-04-01

    ... heterospecific antibodies, and additional tests for nonspecific properties. 660.54 Section 660.54 Food and Drugs..., specificity tests, tests for heterospecific antibodies, and additional tests for nonspecific properties. The...) Specificity tests, tests for heterospecific antibodies, and additional tests for nonspecific properties....

  7. Antinuclear antibody testing: discordance between commercial laboratories.

    Science.gov (United States)

    Abeles, Aryeh M; Gomez-Ramirez, Manuel; Abeles, Micha; Honiden, Shyoko

    2016-07-01

    Antinuclear antibody (ANA) test results frequently affect the course of patients' evaluations, diagnosis, and treatment, but different laboratory centers may yield conflicting results. This study investigated the degree of agreement between laboratory results in a group of subjects who had ANA testing performed at two commercial laboratories. This was a chart review study, in which all ANA tests ordered by the authors from one commercial laboratory over a 4-year period were queried. Corresponding patient charts were reviewed, and if ANA testing had also been performed at the second commercial laboratory, subjects were entered into the study. The primary measurement was agreement between paired ANA results, and we performed sensitivity analysis using varying criteria defining agreement (criteria A to criteria D [strictest to most lenient definition of agreement]). Other data captured included relevant data obtained through the course of evaluation (e.g., presenting complaints, exam findings, other laboratory data) and final diagnoses. Of 101 paired ANA tests, there was 18 % agreement according to the strictest criteria and 42 % according to the most lenient. Of the seven subjects with ANA-associated rheumatic disease, none of the paired tests were in agreement according to criteria A (two agreed according to criteria D). Our findings demonstrate poor agreement between paired ANA tests performed at two commercial laboratories. The low level of agreement may have far-reaching clinical implications. Specifically, this finding calls into question the reliability of ANA testing as it is currently performed and suggests that results may in part depend upon the laboratory center to which patients are referred. PMID:27044430

  8. Evaluation of Gamma Interferon and Antibody Tuberculosis Tests in Alpacas

    OpenAIRE

    Rhodes, Shelley; Holder, Tom; Clifford, Derek; Dexter, Ian; Brewer, Jacky; Smith, Noel; Waring, Laura; Crawshaw, Tim; Gillgan, Steve; Lyashchenko, Konstantin; Lawrence, John; Clarke, John; de la Rua-Domenech, Ricardo; Vordermeier, Martin

    2012-01-01

    We describe the performance of cell-based and antibody blood tests for the antemortem diagnosis of tuberculosis (TB) in South American camelids (SAC). The sensitivity and specificity of the gamma interferon (IFN-γ) release assay, two lateral flow rapid antibody tests (Stat-Pak and Dual Path Platform [DPP]), and two enzyme-linked immunosorbent assay (ELISA)-based antibody tests (Idexx and Enferplex) were determined using diseased alpacas from Mycobacterium bovis culture-confirmed breakdown her...

  9. Development of monoclonal antibodies suitable for rabies virus antibody and antigen detection.

    Science.gov (United States)

    Chander, Vishal; Singh, R P; Verma, P C

    2012-12-01

    The control of an infectious viral disease as rabies is made easier by rapid and accurate diagnosis. Successful rabies prophylaxis is dependent upon the active immunization with vaccine along with passive administration of rabies virus neutralizing antibodies which together clear the virus before widespread infection of central nervous system occurs. The present study aimed at the development of monoclonal antibodies (MAbs) suitable for rabies virus antibody and antigen detection. For the production of rabies specific MAbs, immunization of Swiss albino mice with a commercially available vaccine was done and Polyethylene glycol mediated fusion of spleenocytes with myeloma cells was performed. The positive clones were selected on the basis of distinct reactivity by cell Enzyme linked immunosorbent assay and fluorescence in Indirect Fluorescent antibody test. The positive clones obtained were subjected to single cell cloning by limiting dilution method. The reactive clones were further titrated and employed for virus titration and virus neutralization. The neutralizing activity was evaluated using Fluorescence Activated Cell Sorter technique. Three MAb clones showed a distinct percent inhibition in the presence of positive serum. One of the MAb clone No. 5C3 was relatively more specific in detecting rabies antibodies and also found suitable for competitive ELISA to assess the antibody level in vaccinated subjects. PMID:24293819

  10. Modified indirect hemagglutination test for detection of treponemal antibodies in finger-prick blood.

    OpenAIRE

    Backhouse, J L; Lee, M. H.; Nesteroff, S I; Hudson, B J; Hamilton, P. A.

    1992-01-01

    A modified indirect hemagglutination test for the detection of treponemal antibodies was developed for use with finger-prick blood. By using paired serum and absorbed finger-prick blood from 58 patients from an area previously endemic for yaws and 12 patients without yaws, the modified hemagglutination test was compared with a hemagglutination test for Treponema pallidum and the fluorescent treponemal antibody absorption test. The modified hemagglutination test showed 100% specificity and an ...

  11. Recent developments in monoclonal antibody radiolabeling techniques

    International Nuclear Information System (INIS)

    Monoclonal antibodies (MAbs) have shown the potential to serve as selective carriers of radionuclides to specific in vivo antigens. Accordingly, there has been an intense surge of research activity in an effort to develop and evaluate MAb-based radiopharmaceuticals for tumor imaging (radioimmunoscintigraphy) and therapy (radioimmunotherapy), as well as for diagnosing nonmalignant diseases. A number of problems have recently been identified, related to the MAbs themselves and to radiolabeling techniques, that comprise both the selectivity and the specificity of the in vivo distribution of radiolabeled MAbs. This paper will address some of these issues and primarily discuss recent developments in the techniques for radiolabeling monoclonal antibodies that may help resolve problems related to the poor in vivo stability of the radiolabel and may thus produce improved biodistribution. Even though many issues are identical with therapeutic radionuclides, the discussion will focus mainly on radioimmunoscintigraphic labels. 78 refs., 6 tabs

  12. Drug Development of Therapeutic Monoclonal Antibodies.

    Science.gov (United States)

    Mould, Diane R; Meibohm, Bernd

    2016-08-01

    Monoclonal antibodies (MAbs) have become a substantial part of many pharmaceutical company portfolios. However, the development process of MAbs for clinical use is quite different than for small-molecule drugs. MAb development programs require careful interdisciplinary evaluations to ensure the pharmacology of both the MAb and the target antigen are well-understood. Selection of appropriate preclinical species must be carefully considered and the potential development of anti-drug antibodies (ADA) during these early studies can limit the value and complicate the performance and possible duration of preclinical studies. In human studies, many of the typical pharmacology studies such as renal or hepatic impairment evaluations may not be needed but the pharmacokinetics and pharmacodynamics of these agents is complex, often necessitating more comprehensive evaluation of clinical data and more complex bioanalytical assays than might be used for small molecules. This paper outlines concerns and strategies for development of MAbs from the early in vitro assessments needed through preclinical and clinical development. This review focuses on how to develop, submit, and comply with regulatory requirements for MAb therapeutics. PMID:27342605

  13. HIV抗体纳米免疫磁珠快速检测试剂的研制%Development of a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method

    Institute of Scientific and Technical Information of China (English)

    杨发青; Tony Lee; 王朝南; 孙树叶; 李姗姗; 田辉

    2010-01-01

    目的 研制一种用纳米免疫磁珠层析技术用以检测HIV抗体的快速诊断试剂.方法 运用碳二亚(EDC)将重组的HIV抗原gp41、gp36偶联到200 nm的超顺磁纳米颗粒上,在硝酸纤维素(NC)膜上包被gp41、gp36抗原,制备成免疫层析检测卡,然后对检测卡进行性能分析评估.结果 对HIV抗体国家参考品血清盘(胶体金类)检测,符合要求;对20份HIV抗体阳性和600份抗体阴性临床血清检测,灵敏度为100%,特异性为98.5%.检测卡室温保存12个月性能稳定.结论 研制出一种纳米免疫磁珠HIV抗体检测试剂,具有检测简便、快速、稳定性好和适于现场检测等特点.%Objective To develop a rapid test kit for antibody to HIV by nano immunomagnetic lateral flow method.Methods A rapid test kit was developed by conjugation of the HIV antigen gp41 and gp36 to 200nm super paramagnetic particles by carbodiimide (EDC) and coating of the HIV antigen sp41 and gp36 to nitrocellulose membrane.Then the kit was evaluated with serials of experiments.Results The kit was qualified with examination of national reference panel of anti-HIV antibody for colloidal gold diagnostic kit.The sensitivity was 100% by tested with 20 HIV antibody positive sera,the specificity was 98.5% by tested with 600 HIV antibody negative sera,respectively.The stabilitv of the kit was over 12 month by storage at room temperature.Conclusion A diagnostic kit for antibody to HIV was developed with the advantages of convenience,rapid test,good stability and point of care.

  14. Development of Scoring Functions for Antibody Sequence Assessment and Optimization

    OpenAIRE

    Seeliger, Daniel

    2013-01-01

    Antibody development is still associated with substantial risks and difficulties as single mutations can radically change molecule properties like thermodynamic stability, solubility or viscosity. Since antibody generation methodologies cannot select and optimize for molecule properties which are important for biotechnological applications, careful sequence analysis and optimization is necessary to develop antibodies that fulfil the ambitious requirements of future drugs. While efforts to gra...

  15. Triiodothyronine uptake test using specific antibody as the secondary binder

    International Nuclear Information System (INIS)

    We describe a specific and reliable triiodithyronine uptake (T3-U) method for the estimation of free thyroxine index(FT4I) using precipitated anti-T3-antibody and second-antibody (anti-rabbit-IgG raised in goat) complex. Since the method measures the partitioning of T3-125I between the binding proteins and antibody, separation of the portion of T3-125I taken up by the antibody from that bound to the binding proteins is achieved by the use of pre-incubated primary and second antibody complex in the form of suspension, and separation of the bound complex was carried out by means of centrifugation. The T3-U method developed is clinically evaluated and compared with the reputed commercial kit and a correlation of 0.96 was obtained. (author)

  16. Immunoglobulins, antibody repertoire and B cell development.

    Science.gov (United States)

    Butler, J E; Zhao, Y; Sinkora, M; Wertz, N; Kacskovics, I

    2009-03-01

    Swine share with most placental mammals the same five antibody isotypes and same two light chain types. Loci encoding lambda, kappa and Ig heavy chains appear to be organized as they are in other mammals. Swine differ from rodents and primates, but are similar to rabbits in using a single VH family (VH3) to encode their variable heavy chain domain, but not the family used by cattle, another artiodactyl. Distinct from other hoofed mammals and rodents, Ckappa:Clambda usage resembles the 1:1 ratio seen in primates. Since IgG subclasses diversified after speciation, same name subclass homologs do not exist among swine and other mammals unless very closely related. Swine possess six putative IgG subclasses that appear to have diversified by gene duplication and exon shuffle while retaining motifs that can bind to FcgammaRs, FcRn, C1q, protein A and protein G. The epithelial chorial placenta of swine and the precosial nature of their offspring have made piglets excellent models for studies on fetal antibody repertoire development and on the postnatal role of gut colonization, maternal colostrum and neonatal infection on the development of adaptive immunity during the "critical window" of immunological development. This chapter traces the study of the humoral immune system of this species through its various eras of discovery and compiles the results in tables and figures that should be a useful reference for educators and investigators. PMID:18804488

  17. Lectin immuno tests: quantitation and titration of antigens and antibodies using lectin-antibody conjugates

    International Nuclear Information System (INIS)

    The authors have investigated the possibility of using lectin-antibody conjugates as general reagents in immunological procedures requiring a labeled antigen or antibody. Using these conjugates, labeling is achieved through saccharide binding sites of lectins which operate as acceptors for glycoconjugate marker substances added secondarily. Marker substances used in this work were enzymes, radioactively labeled glycoconjugates and erythrocytes, but other markers can also be used. Using the first two markers, antigens and antibodies were determined with accuracy and sensitivity equal to those of conventional enzyme or radioimmunoassays. Using erythrocytes as a marker, a simple erythro-adsorption procedure, possibly followed by hemolysis, has been developed which allowed the titration of antigens and antibodies to be carried out with a sensitivity at least equal to enzyme or radioimmunoassays. (Auth.)

  18. Development of Monoclonal Antibodies Suitable for Rabies Virus Antibody and Antigen Detection

    OpenAIRE

    Chander, Vishal; Singh, R.P.; Verma, P. C.

    2012-01-01

    The control of an infectious viral disease as rabies is made easier by rapid and accurate diagnosis. Successful rabies prophylaxis is dependent upon the active immunization with vaccine along with passive administration of rabies virus neutralizing antibodies which together clear the virus before widespread infection of central nervous system occurs. The present study aimed at the development of monoclonal antibodies (MAbs) suitable for rabies virus antibody and antigen detection. For the pro...

  19. ANCA / MPO / PR3 Antibodies Test

    Science.gov (United States)

    ... the cells. The sample is put on a slide and treated with a fluorescent stain. The slide is then examined under a microscope and the ... prior to or along with ANCA testing to rule out other causes for the symptoms. ^ Back to ...

  20. Comparison of commercial diagnostic tests for Helicobacter pylori antibodies.

    OpenAIRE

    Schembri, M. A.; Lin, S K; Lambert, J R

    1993-01-01

    A number of serological tests measuring the presence of Helicobacter pylori-specific serum immunoglobulin G (IgG) are now commercially available. The aim of this study was to evaluate the clinical accuracy of five commercial H. pylori antibody tests: GAP-IgG (Biomerica), HELpTEST (AMRAD, Kew, Victoria, Australia), HELICO-G (Porton Cambridge), Pyloriset (Orion Diagnostica), and ROCHE (Roche Diagnostics). A total of 162 subjects presenting for routine upper endoscopy were studied. H. pylori was...

  1. The Problem of Thyroid Antibodies Testing

    OpenAIRE

    Lukinac, Ljerka; Krilić, Dražena; Nöthig-Hus, Dunja; Kusić, Zvonko

    2004-01-01

    Nowadays, different methods for determination of thyroglobulin autoantibodies (TGA), microsomal autoantibodies (TMA) and autoantibodies to enzyme thyroid peroxidase (TPO) have been developed. The specificity and sensitivity of these methods depend on the purity of autoantigen preparation itself, valid standardization and type of the methodology used, e.g., agglutination of gelatine particle carriers sensitized with antigen, radioimmunoassay (RIA), immunometric assay (IRMA), enzyme immunoassay...

  2. Detection of IgE insulin antibody with radioallergosorbent test

    International Nuclear Information System (INIS)

    An in vitro method for detecting IgE insulin antibody using the principle of the radioallergosorbent test (RAST) is described. In six patients with insulin allergy, the RAST values were higher than in normal persons or insulin-treated diabetics without insulin allergy. No differences were observed between normal persons and insulin-treated diabetics without insulin allergy. Moreover, it was observed that in one patient treated with highly purified insulin, there was a gradual decrease of RAST value parallel to the radioinsulin binding activity and clinical allergic symptoms. The RAST value of insulin is slightly inhibited by non-IgE antibodies and is, therefore, a semiquantitative value. However, the RAST is simple to perform and reproducible; it is therefore very useful in the detection of IgE insulin antibodies. (orig.)

  3. Monoclonal Antibody Testing for Cancer Metastasis

    Science.gov (United States)

    1993-01-01

    ) extracted from breast tumor tissues have recently been shown, together with plasminogen activator inhibitor 1 (PAI-1), to be a good prognostic indicator for high risk of recurrence and shorter patient survival times. In this project, we have attempted to develop immunocytochemical methodologies for the clinical assessment of the expression of urokinase plasminogen activator, which has been implicated to be important for initial steps in tumor invasion, and to relate it to cell proliferation and DNA replication at the single-cell level.

  4. 一种基于量子点检测抗CCP抗体的免疫荧光层析法%Development of an Immunochromatographic Test Strip for the Detection of Anti-CCP Antibody Based on Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    郭利宁; 何红秋

    2013-01-01

    抗环瓜氨酸多肽(cyclic citrullinated peptide,CCP)抗体是类风湿关节炎(rheumatoid arthritis,RA)早期诊断的重要生物标志物.为了实现对RA的早期诊断,本研究建立了一种基于CdTe量子点标记技术检测抗CCP抗体的免疫荧光层析法.将CCP多肽与小牛血清白蛋白(bovine serum albumin,BSA)连接,再将CCP-BSA和羊抗鼠IgG分别在硝酸纤维素膜(nitrocellulose membrane,NC膜)上划线,作为检测线(test line,T线)和质控线(control line,C线).制备量子点并在量子点上标记鼠抗人IgG,喷在玻璃纤维上并烘干,最后组装大卡、切割并封装制成检测试纸条.应用该试纸条检测了RA患者及健康人血清临床样本200份,以酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)为对照,计算免疫荧光层析法的检测灵敏度和特异性.结果显示,建立的量子点免疫荧光层析试纸条检测抗CCP抗体的灵敏度为97.5%,特异性为95.8%.该方法操作简单、快速,可实现床旁检测(point-of-care testing,POCT),能应用于RA的早期诊断.%Anti-cyclic citrullinated peptide antibody ( anti-CCP antibody) is an important biomarker for early detection of rheumatoid arthritis ( RA). For clinical diagnosis of RA in the early stage, an immunochromatographic test strip for anti-CCP antibody detection was developed based on quantum dots. First, CCP was linked to bovine serum albumin (BSA) ; and then, the CCP-BSA and goat anti-mouse IgG were dotted on the nitrocellulose membrane as test line ( T line) and control line ( C line) , respectively. Subsequently, quantum dots solution was prepared and conjugated to the mouse anti-human IgG antibody; further more, the marked quantum dots were sprayed to the glass fibers and dried. Last, all components of immunochromatographic test strip were assembled, cut and packaged. The test strip has been applied to the detection of 200 clinic serum samples from RA patients and healthy people, and the sensitivity

  5. Development of antibodies to human embryonic stem cell antigens

    OpenAIRE

    Stanley Marisa; Rao Mahendra S; Olson Judith M; Cai Jingli; Taylor Eva; Ni Hsiao-Tzu

    2005-01-01

    Abstract Background Using antibodies to specific protein antigens is the method of choice to assign and identify cell lineage through simultaneous analysis of surface molecules and intracellular markers. Embryonic stem cell research can be benefited from using antibodies specific to transcriptional factors/markers that contribute to the "stemness" phenotype or critical for cell lineage. Results In this report, we have developed and validated antibodies (either monoclonal or polyclonal) specif...

  6. Development and evaluation of rapid point-of-care tests for detection of Enterovirus 71 and Coxsackievirus A16 specific immunoglublin M antibodies.

    Science.gov (United States)

    Zhang, Jing; Weng, Zuxing; Du, Hailian; Xu, Feihai; He, Shuizhen; He, Delei; Cheng, Tong; Zhang, Jun; Ge, Shengxiang; Xia, Ningshao

    2016-05-01

    Two colloidal gold immunochromatographic assays (CGIAs) for detection of EV71- and CA16- immunoglobulin M (IgM) were developed and evaluated. A total of 1465 sera collected from children with hand, foot, and mouth disease (HFMD), non-HFMD patients and healthy children. The sensitivity of IgM CGIA tests for EV71 and CA16 were 97.6% (330/338) and 91.6% (296/323) respectively, compared to those who were viral RNA positive by PCR. Their performances were comparable to those of commercial ELISA kits, with agreement of 98.1% for EV71-IgM and 97.3% for CA16-IgM. In addition, for EV71- and CA16-IgM CGIAs, the results of whole blood samples were 99.6% (248/249) and 100% (191/191) concordant to those with serum samples, respectively. As rapid point-of-care (POC) tests, the two CGIAs were suitable to be used in community clinic units, especially in resource-poor areas and will facilitate the control of HFMD. PMID:26912234

  7. Red Blood Cell Antibodies in Hematology/Oncology Patients: Interpretation of Immunohematologic Tests and Clinical Significance of Detected Antibodies.

    Science.gov (United States)

    Hendrickson, Jeanne E; Tormey, Christopher A

    2016-06-01

    Red blood cell (RBC) transfusion is a cornerstone of the management of patients with hematology/oncology disorders. However, a potentially deleterious consequence of transfusion is the development of alloantibodies against blood group antigens present on RBCs. Such alloantibodies can be an obstacle in providing compatible units for transfusion. Providers in this arena must fully understand the testing performed by blood banks, as well as the consequences of detected antibodies. This article reviews immunohematologic tests, describes how autoimmune hemolytic anemia is classified by autoantibodies; outlines RBC alloimmunization rates, and presents strategies to prevent/mitigate the impact of RBC alloimmunization. PMID:27113001

  8. 21 CFR 866.5785 - Anti-Saccharomyces cerevisiae (S. cerevisiae) antibody (ASCA) test systems.

    Science.gov (United States)

    2010-04-01

    ...) antibody (ASCA) test systems. 866.5785 Section 866.5785 Food and Drugs FOOD AND DRUG ADMINISTRATION... Immunological Test Systems § 866.5785 Anti-Saccharomyces cerevisiae (S. cerevisiae) antibody (ASCA) test systems. (a) Identification. The Anti-Saccharomyces cerevisiae (S. cerevisiae) antibody (ASCA) test system...

  9. Specificity, sensitivity, and reproducibility among the fluorescent treponemal antibody-absorption test, the microhemagglutination assay for Treponema pallidum antibodies, and the hemagglutination treponemal test for syphilis.

    OpenAIRE

    Larsen, S A; Hambie, E A; Pettit, D E; Perryman, M W; Kraus, S J

    1981-01-01

    Using 920 sera, we compared the specificity and reproducibility of the hemagglutination treponemal test for syphilis with those of the fluorescent treponemal antibody-absorption test and the microhemagglutination assay for Treponema pallidum antibodies; we found all three tests to be comparable. However, the hemagglutination treponemal test for syphilis, like the microhemagglutination assay for T. pallidum antibodies, lacked sensitivity in sera from patients with primary syphilis.

  10. Development of monoclonal antibodies that recognize Treponema pallidum.

    OpenAIRE

    Saunders, J. M.; Folds, J D

    1983-01-01

    We developed a panel of monoclonal antibodies to Treponema pallidum (Nichols) antigens, some of which recognize treponemal antigens on T. pallidum (Nichols), T. pallidum strain 14, and Treponema phagedenis biotype Reiter. The antibodies were detected by either an enzyme-linked immunosorbent assay or a radioimmunoassay.

  11. Developing recombinant antibodies for biomarker detection

    Energy Technology Data Exchange (ETDEWEB)

    Baird, Cheryl L.; Fischer, Christopher J.; Pefaur, Noah B.; Miller, Keith D.; Kagen, Jacob; Srivastava, Sudhir; Rodland, Karin D.

    2010-10-01

    Monoclonal antibodies (mAbs) have an essential role in biomarker validation and diagnostic assays. A barrier to pursuing these applications is the reliance on immunization and hybridomas to produce mAbs, which is time-consuming and may not yield the desired mAb. We recommend a process flow for affinity reagent production that utilizes combinatorial protein display systems (eg, yeast surface display or phage display) rather than hybridomas. These systems link a selectable phenotype-binding conferred by an antibody fragment-with a means for recovering the encoding gene. Recombinant libraries obtained from immunizations can produce high-affinity antibodies (<10 nM) more quickly than other methods. Non-immune libraries provide an alternate route when immunizations are not possible, or when suitable mAbs are not recovered from an immune library. Directed molecular evolution (DME) is an integral part of optimizing mAbs obtained from combinatorial protein display, but can also be used on hybridoma-derived mAbs. Variants can easily be obtained and screened to increase the affinity of the parent mAb (affinity maturation). We discuss examples where DME has been used to tailor affinity reagents to specific applications. Combinatorial protein display also provides an accessible method for identifying antibody pairs, which are necessary for sandwich-type diagnostic assays.

  12. Antibody

    Science.gov (United States)

    An antibody is a protein produced by the body's immune system when it detects harmful substances, called antigens. Examples ... microorganisms (bacteria, fungi, parasites, and viruses) and chemicals. Antibodies may be produced when the immune system mistakenly ...

  13. 21 CFR 866.5120 - Antismooth muscle antibody immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antismooth muscle antibody immunological test... Systems § 866.5120 Antismooth muscle antibody immunological test system. (a) Identification. An antismooth muscle antibody immunological test system is a device that consists of the reagents used to measure...

  14. Human immunodeficiency virus antibody test and seroprevalence in psychiatric patients.

    Science.gov (United States)

    Naber, D; Pajonk, F G; Perro, C; Löhmer, B

    1994-05-01

    Psychiatric inpatients are at risk for human immunodeficiency virus (HIV) infection. Investigations in the United States revealed seroprevalence rates of 5.5-8.9%. Therefore, inclusion of HIV antibody testing in routine laboratory screening is sometimes suggested. To investigate this issue for inpatients in the Department of Psychiatry, University of Munich, the incidence, reason for HIV testing and results were analyzed. Of 12,603 patients, hospitalized from 1985 to 1993, 4.9% (623 patients, 265 in risk groups) underwent the HIV test after informed consent. Thirty patients (4.8% of those tested) were found to be positive, but only in 5 cases (all of risk groups) was infection newly detected. Data indicate that, in psychiatry, HIV testing is reasonable only in patients in risk groups or if clinical variables suggest HIV infection. PMID:8067276

  15. A latex slide agglutination test for rapid detection of antimyeloperoxidase antibody.

    OpenAIRE

    Ko, K.H.; S. S. Lee; Lawton, J W

    1999-01-01

    AIM: To develop and test a new latex slide agglutination test (MPO-LSAT) to detect antimyeloperoxidase (anti-MPO) antibody in serum. METHODS: Latex bead coating was adjusted to give maximum sensitivity by attending to latex size, MPO to latex ratio for coupling, ratio of diluted serum to MPO-latex, reaction time and temperature for coupling, and reaction time for agglutination. Inhibition studies were performed using MPO, proteinase 3, bactericidal/permeability increasing protein, and lactofe...

  16. 21 CFR 864.9175 - Automated blood grouping and antibody test system.

    Science.gov (United States)

    2010-04-01

    ...) Identification. An automated blood grouping and antibody test system is a device used to group erythrocytes (red blood cells) and to detect antibodies to blood group antigens. (b) Classification. Class II (performance... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated blood grouping and antibody test...

  17. Development and characterization of antibody reagents for detecting nanoparticles

    OpenAIRE

    Ravichandran, Supriya; Sullivan, Mark A.; Callahan, Linda M.; Bentley, Karen L.; DeLouise, Lisa A.

    2015-01-01

    The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in sol...

  18. Interim Guidance for Interpretation of Zika Virus Antibody Test Results.

    Science.gov (United States)

    Rabe, Ingrid B; Staples, J Erin; Villanueva, Julie; Hummel, Kimberly B; Johnson, Jeffrey A; Rose, Laura; Hills, Susan; Wasley, Annemarie; Fischer, Marc; Powers, Ann M

    2016-01-01

    Zika virus is a single-stranded RNA virus in the genus Flavivirus and is closely related to dengue, West Nile, Japanese encephalitis, and yellow fever viruses (1,2). Among flaviviruses, Zika and dengue virus share similar symptoms of infection, transmission cycles, and geographic distribution. Diagnostic testing for Zika virus infection can be accomplished using both molecular and serologic methods. For persons with suspected Zika virus disease, a positive real-time reverse transcription-polymerase chain reaction (rRT-PCR) result confirms Zika virus infection, but a negative rRT-PCR result does not exclude infection (3-7). In these cases, immunoglobulin (Ig) M and neutralizing antibody testing can identify additional recent Zika virus infections (6,7). However, Zika virus antibody test results can be difficult to interpret because of cross-reactivity with other flaviviruses, which can preclude identification of the specific infecting virus, especially when the person previously was infected with or vaccinated against a related flavivirus (8). This is important because the results of Zika and dengue virus testing will guide clinical management. Pregnant women with laboratory evidence of Zika virus infection should be evaluated and managed for possible adverse pregnancy outcomes and be reported to the U.S. Zika Pregnancy Registry or the Puerto Rico Zika Active Pregnancy Surveillance System for clinical follow-up (9,10). All patients with clinically suspected dengue should have proper management to reduce the risk for hemorrhage and shock (11). If serologic testing indicates recent flavivirus infection that could be caused by either Zika or dengue virus, patients should be clinically managed for both infections because they might have been infected with either virus. PMID:27254248

  19. Development and maturation of norovirus antibodies in childhood.

    Science.gov (United States)

    Blazevic, Vesna; Malm, Maria; Honkanen, Hanna; Knip, Mikael; Hyöty, Heikki; Vesikari, Timo

    2016-04-01

    The burden of norovirus (NoV) gastroenteritis is substantial in young children. Maternal antibodies are thought to protect a child from NoV infection in early infancy but subsequent development of NoV-specific protective immunity in children is still largely unexplored. We have determined NoV-specific antibody seroconversion to GII.4 virus-like particles as an indicator of NoV infection in two children prospectively followed from birth to eight years of age. Blocking activity and affinity maturation of maternal and serum IgG antibodies were evaluated. Our results show that multiple infections occur in children up to eight years of age. The titer, blocking activity and avidity of maternal antibodies determined susceptibility of an infant to NoV infection. NoV GII.4-specific antibodies with high blocking potential and avidity were developed at two to three years of age and were retained throughout the follow-up. Subsequent NoV infections may have contributed to the duration of protective NoV-specific immune responses that lasted for several years. This study adds to current understanding of the duration of passive protection by maternal antibodies and the duration and quality of acquired immunity following primary and subsequent NoV infections in infants and young children, who are the main target group for NoV vaccine development. PMID:26724451

  20. A population screening test for antibody to measles virus

    International Nuclear Information System (INIS)

    In areas where sporadic cases of measles continue to occur in spite of vaccination programs, the availability of a simple screening test for determination of seropositivity to measles virus is desirable. A sensitive radioimmunoassay (RIA) screening test (ST) for the detection of IgG antibody to measles virus, based on a solid phase RIA, is described. The assays were performed on polyvinyl microtiter plates for which the RIAST requires only 5 μl of serum per subject. Antigen consisted of a sonicated extract of measles virus-infected Vero cells. Rabbit antihuman IgG specific for the Fc-segment of human IgG, labelled with 125I, was used to detect human IgG bound to viral antigen. The basic RIA method was characterized by carrying out full titrations of sera of 53 healthy adults, 10 children, and 13 patients with measles-associated illness. These sera were also tested by the hemagglutination inhibition (HI) technique; most of the measles sera were also tested by complement fixation (CF). RIAST results (expressed as binding ratios) obtained for 52 healthy adults are compared with their RIA serum titers. Of the 200 sera of patients of various ages tested by the RIAST, 63 borderline sera were also tested by HI. The RIAST, which does not require serum treatment other than inactivation, proved to be more sensitive as an indicator of seropositivity than HI. Implications of the results and practical applications of the screening test are discussed. (author)

  1. Voluntary HIV antibody testing amongst youth in Hong Kong.

    Science.gov (United States)

    Lam, Tai Hing; Janghorbani, Mohsen; Fan, Susan; Fielding, Richard

    2003-02-01

    We performed a study to estimate the prevalence of and factors associated with human immunodeficiency virus (HIV) antibody-testing behaviour among youth in Hong Kong. It was a population-based cross-sectional study. Questions on HIV testing were asked as part of a youth sexuality study conducted in July to December 1996 among young adults in Hong Kong. A total of 517 (53.6%) males and 447 (46.4%) females aged 17 to 28 years completed an anonymous structured self-administered questionnaire. Respondents had good knowledge about correct modes of HIV transmission and prevention. 9.4% (95% confidence interval [CI]: 7.0, 12.3) of males and 6.4% (95% CI: 4.3, 9.1) of females had been tested for HIV through blood donation. Excluding blood donation, 3.7% (95% CI: 2.2, 5.7) of males and 3.6% (95% CI: 2.1, 5.9) of females had been tested (voluntary testing). 47.5% (95% CI: 44.4, 50.7) of subjects reported at least one major risk factor for HIV infection. In multivariate analyses, factors independently associated with both voluntary HIV testing and HIV testing by blood donation were age and having had sex with multiple partners. A higher educational level was a predictor of HIV testing by blood donation. Self-assessment of having sufficient sex education was also significantly associated with voluntary HIV testing. HIV testing is not widespread in Hong Kong and those at risk are more likely to have been tested. It is of concern, however, that many of those reporting risk factors have not been tested. PMID:12662393

  2. Developments in the production of mucosal antibodies in plants.

    Science.gov (United States)

    Vasilev, Nikolay; Smales, C Mark; Schillberg, Stefan; Fischer, Rainer; Schiermeyer, Andreas

    2016-01-01

    Recombinant mucosal antibodies represent attractive target molecules for the development of next generation biopharmaceuticals for passive immunization against various infectious diseases and treatment of patients suffering from mucosal antibody deficiencies. As these polymeric antibodies require complex post-translational modifications and correct subunit assembly, they are considered as difficult-to-produce recombinant proteins. Beside the traditional, mammalian-based production platforms, plants are emerging as alternative expression hosts for this type of complex macromolecule. Plant cells are able to produce high-quality mucosal antibodies as shown by the successful expression of the secretory immunoglobulins A (IgA) and M (IgM) in various antibody formats in different plant species including tobacco and its close relative Nicotiana benthamiana, maize, tomato and Arabidopsis thaliana. Importantly for biotherapeutic application, transgenic plants are capable of synthesizing functional IgA and IgM molecules with biological activity and safety profiles comparable with their native mammalian counterparts. This article reviews the structure and function of mucosal IgA and IgM antibodies and summarizes the current knowledge of their production and processing in plant host systems. Specific emphasis is given to consideration of intracellular transport processes as these affect assembly of the mature immunoglobulins, their secretion rates, proteolysis/degradation and glycosylation patterns. Furthermore, this review provides an outline of glycoengineering efforts that have been undertaken so far to produce antibodies with homogenous human-like glycan decoration. We believe that the continued development of our understanding of the plant cellular machinery related to the heterologous expression of immunoglobulins will further improve the production levels, quality and control of post-translational modifications that are 'human-like' from plant systems and enhance the

  3. Development and standardization of multiplexed antibody microarrays for use in quantitative proteomics

    Directory of Open Access Journals (Sweden)

    Sorette M

    2004-12-01

    Full Text Available Abstract Background Quantitative proteomics is an emerging field that encompasses multiplexed measurement of many known proteins in groups of experimental samples in order to identify differences between groups. Antibody arrays are a novel technology that is increasingly being used for quantitative proteomics studies due to highly multiplexed content, scalability, matrix flexibility and economy of sample consumption. Key applications of antibody arrays in quantitative proteomics studies are identification of novel diagnostic assays, biomarker discovery in trials of new drugs, and validation of qualitative proteomics discoveries. These applications require performance benchmarking, standardization and specification. Results Six dual-antibody, sandwich immunoassay arrays that measure 170 serum or plasma proteins were developed and experimental procedures refined in more than thirty quantitative proteomics studies. This report provides detailed information and specification for manufacture, qualification, assay automation, performance, assay validation and data processing for antibody arrays in large scale quantitative proteomics studies. Conclusion The present report describes development of first generation standards for antibody arrays in quantitative proteomics. Specifically, it describes the requirements of a comprehensive validation program to identify and minimize antibody cross reaction under highly multiplexed conditions; provides the rationale for the application of standardized statistical approaches to manage the data output of highly replicated assays; defines design requirements for controls to normalize sample replicate measurements; emphasizes the importance of stringent quality control testing of reagents and antibody microarrays; recommends the use of real-time monitors to evaluate sensitivity, dynamic range and platform precision; and presents survey procedures to reveal the significance of biomarker findings.

  4. Microcapsule agglutination test for Treponema pallidum antibodies. A new serodiagnostic test for syphilis.

    OpenAIRE

    Kobayashi, S; Yamaya, S I; Sugahara, T.; Matuhasi, T.

    1983-01-01

    For the serodiagnosis of syphilis a quantitative passive agglutination (MCA-TP) test for antibodies to Treponema pallidum was performed with chemically stable microcapsules with no antigenic activity instead of with conventional sheep erythrocytes. The microcapsules were easily sensitised with the antigen of sonicated Treponema pallidum by treatment with glutaraldehyde. Compared with the Treponema pallidum haemagglutination test (TPHA) the MCA-TP test was superior for detecting cases of prima...

  5. IgE antibody responses in schistosomiasis measured by a radioallergosorbent test

    International Nuclear Information System (INIS)

    Helminth infections are associated with the production of unusually high concentrations of circulating IgE antibody. Assays for IgE antibodies should provide useful approaches for the study of protective immunity and may also be of use in serodiagnosis of diseases induced by helminths. The radioallergosorbent test is carried out by attaching the antigen or allergen to an insoluble supportive material, allowing the IgE antibodies in the test serum to react with excess bound antigen, and then estimating the IgE antibody bound by its reaction with 125I-labelled goat anti-human IgE antibody

  6. Development of a monoclonal antibody specific to cooked mammalian meats.

    Science.gov (United States)

    Hsieh, Y H; Sheu, S C; Bridgman, R C

    1998-04-01

    Detection of species adulteration in ground meat products is important for consumer protection and food-labeling law enforcement. This study was conducted to develop monoclonal antibodies (MAbs) that can be used in an enzyme-linked immunosorbent assay (ELISA) for rapid detection of any cooked mammalian meats in cooked poultry products. Soluble muscle proteins extracted from cooked pork (heated at 100 degrees C for 15 min) were used as the antigen to immunized mice for developing the MAb. One that was developed, MAb 2F8 (IgG2b class), strongly reacted with cooked meat of five mammalian species (beef cattle, hogs, sheep, horse, and deer) but did not react with any cooked poultry (chicken, turkey, and duck) or raw meats. At least 0.5% by weight of pork, beef, lamb, and horse meats in a chicken-based mixture could not detect using the indirect ELISA with MAb 2F8. The MAb 2F8 is useful in a single initial screening test to detect the presence of five nonpoultry meat adulterants in cooked poultry products. PMID:9709213

  7. Indirect micro-immunofluorescence test for detecting type-specific antibodies to herpes simplex virus.

    OpenAIRE

    Forsey, T; Darougar, S

    1980-01-01

    A rapid indirect micro-immunofluorescence test capable of detecting and differentiating type-specific antibodies to herpes simplex virus is described. The test proved highly sensitive and, in 80 patients with active herpes ocular infection, antibody was detected in 94%. No anti-herpes antibody was detected in a control group of 20 patients with adenovirus infections. Testing of animal sera prepared against herpes simplex virus types 1 and 2 and of human sera from cases of ocular and genital h...

  8. Anticomplement immunofluorescence test that uses isolated fibroblast nuclei for detection of antibodies to human cytomegalovirus.

    OpenAIRE

    Mintz, L; Miner, R. C.; Yeager, A S

    1980-01-01

    Cytomegalovirus antibodies were measured in human sera by a nuclear anticomplement immunofluorescence test that used as antigen the isolated nucleic of virus-infected fibroblasts cells lysed in distilled water. The method exhibited less nonspecific fluorescence than either a conventional whole-cell anticomplement immunofluorescence test or an indirect fluorescent antibody test applied to the same isolated nuclear substrate. The assay detected 97.5% of 40 antibody-positive sera, compared with ...

  9. Micro-indirect hemagglutination test for detection of antibodies to the Ibc protein of group B Streptococcus.

    OpenAIRE

    Thangavelu, C P; Koshi, G

    1980-01-01

    A micro-indirect hemagglutination test was developed for detecting antibody against the Ibc protein of group B Streptococcus. Formalin-preserved, tanned sheep erythrocytes were sensitized with a partially purified preparation of Ibc protein from a type Ic strain of group B streptococci. A total of 76% of 103 sera from pregnant and nonpregnant women had demonstrable antibody against this protein, with titers ranging from 10 to 320. Examination of five pairs of mother and cord sera revealed pas...

  10. Anti Rh Hemolytic Disease due to Anti C Antibody: Is Testing for Anti D Antibodies Enough?

    OpenAIRE

    Negi, Gita; Singh, Gaur Dushyant

    2011-01-01

    Rh blood group system is a complex blood group system. Rh antibodies are produced in Rh negative individuals following exposure to foreign RBCs after transfusion or pregnancy. Anti C is a rare cause of hemolytic disease of newborn and is very scarcely reported in the literature. The aim of the present case report of Hemolytic disease caused by Anti C antibody is to bring out the fact that antibodies other than anti D should be considered in cases that give a suggestive history but no evidence...

  11. Development of an EGFRvIII specific recombinant antibody

    Directory of Open Access Journals (Sweden)

    Li Gordon

    2010-10-01

    Full Text Available Abstract Background EGF receptor variant III (EGFRvIII is the most common variant of the EGF receptor observed in human tumors. It results from the in frame deletion of exons 2-7 and the generation of a novel glycine residue at the junction of exons 1 and 8. This novel juxtaposition of amino acids within the extra-cellular domain of the EGF receptor creates a tumor specific and immunogenic epitope. EGFRvIII expression has been seen in many tumor types including glioblastoma multiforme (GBM, breast adenocarcinoma, non-small cell lung carcinoma, ovarian adenocarcinoma and prostate cancer, but has been rarely observed in normal tissue. Because this variant is tumor specific and highly immunogenic, it can be used for both a diagnostic marker as well as a target for immunotherapy. Unfortunately many of the monoclonal and polyclonal antibodies directed against EGFRvIII have cross reactivity to wild type EGFR or other non-specific proteins. Furthermore, a monoclonal antibody to EGFRvIII is not readily available to the scientific community. Results In this study, we have developed a recombinant antibody that is specific for EGFRvIII, has little cross reactivity for the wild type receptor, and which can be easily produced. We initially designed a recombinant antibody with two anti-EGFRvIII single chain Fv's linked together and a human IgG1 Fc component. To enhance the specificity of this antibody for EGFRvIII, we mutated tyrosine H59 of the CDRH2 domain and tyrosine H105 of the CDRH3 domain to phenylalanine for both the anti-EGFRvIII sequence inserts. This mutated recombinant antibody, called RAbDMvIII, specifically detects EGFRvIII expression in EGFRvIII expressing cell lines as well as in EGFRvIII expressing GBM primary tissue by western blot, immunohistochemistry (IHC and immunofluorescence (IF and FACS analysis. It does not recognize wild type EGFR in any of these assays. The affinity of this antibody for EGFRvIII peptide is 1.7 × 107 M-1 as

  12. Development and monitoring of a novel monoclonal antibody purification strategy

    OpenAIRE

    Capito, Florian

    2014-01-01

    The studies presented in the cumulative part of this thesis illustrate the different steps to develop a polymer-driven antibody purification process. These peer-reviewed reports show in detail fundamental research, additional method development useful in the development of such a purification process as well as implementation of the final process. A strategy for analyzing copolymers, synthesized by a lab in house, was implemented with particular emphasis on copolymer composition analysis. Thi...

  13. Immunological considerations for developing antibody therapeutics for Influenza A.

    Science.gov (United States)

    Chan-Hui, Po-Ying; Swiderek, Kristine M

    2016-02-01

    Influenza infection can give rise to serious illness leading to complications and hospitalization of patients. The efficacy of current standard of care is very limited and provides little relief for patients hospitalized with serious flu. Human monoclonal antibodies (mAb) against influenza are being developed as new treatment options for this patient population. When developing antibody therapeutics, it is important to consider all possible immunologic effects of the antibodies on viral infection and disease progression including those other than the postulated therapeutic mechanisms. An area of concern is the potential of antibody-dependent enhancement (ADE) of illness. ADE of viral infections has been extensively described for Dengue virus (DENV) but not for influenza. Recently, preliminary results from clinical viral challenge studies of anti-HA-stalk mAbs suggested the possibility of enhanced viral shedding, raising concerns for ADE when utilizing mAbs as therapeutic intervention for influenza although viral shedding was not enhanced in the clinical viral challenge of anti-M2 mAb TCN-032. We herein discuss the known mechanisms of ADE and their relevance to developing mAbs such as anti-HA and anti-M2 for influenza disease. PMID:26325257

  14. Development of radioactivity labelling method of new antibody by using the antibody engineering

    International Nuclear Information System (INIS)

    With an aim to develop a method to produce labelled antibodies with low immunogenicity, two recombinant fusion proteins; scFv-His and scFv-MTβ were produced using gene engineering techniques. The former was constructed with scFv-antibody and histidine hexamer, a metal-chelated protein (or peptide). The latter was done with scFv-antibody and β-domain of metallothionein. Then, antigen-binding activity and metal-binding activity of these fusion proteins were determined using gel-filtration chromatography and ELISA. The main antigen-binding activity of scFv-His preparation was detected in a domain of about 25-30 kDa, which agreed with the peak of 29 kDa corresponding to the presumed molecular weight for the protein. Whereas the antigen-binding activity of scFv-MTβ was found in a domain of 30-35 kDa, which agreed with 32 kDa, the presumed molecular weight of scFv-MTβ. Gel-filtration chromatography of scFv-His preparation after the addition of Cu2+ ion revealed an optical absorption at 280 nm and a Cu-peak near at 14 kDa. These results suggested that the metal affinity of the histidine-hexamer was too weak to chelate Cu2+ in a solution. The chromatography of scFv-MTβ preparation added with Cd2+ showed a peak of Cd appeared around a position of about 20 kDa but the peak was not coincident with that of the antigen-binding activity (ca. 30 kDa), suggesting that the present preparation of scFv-MTβ had no Cd-binding activity due to metal-exchange reaction. Based on these results, problems on the production of recombinant scFv-antibody fused with metal-binding domain of cystein-binding type or histidine-binding one were discussed. (M.N.)

  15. Development of Monoclonal Antibodies in China: Overview and Prospects

    OpenAIRE

    2015-01-01

    Monoclonal antibodies (mAbs) have become increasingly important as human therapeutic agents. Yet, current research concentrates on technology itself and pays attention to developed countries. This paper aims to provide a comprehensive review of mAbs development in China through systematic analysis of drug registry, patent applications, clinical trials, academic publication, and ongoing R&D projects. The trends in therapeutic areas and industrialization process are also highlighted. Developmen...

  16. Comparison of an enzyme-linked immunoassay and a quantitative indirect fluorescent-antibody test with the conventional indirect fluorescent-antibody test for detecting antibodies to Toxoplasma gondii.

    OpenAIRE

    Violand, S A; Mitchell, T G; Kleeman, K T

    1982-01-01

    Two new methods for the detection of antibodies to Toxoplasma gondii, an enzyme-linked immunosorbent assay and a quantitative immunofluorescence assay, were evaluated and compared with the conventional indirect fluorescent-antibody slide test. Each of 100 human sera was assayed twice by the three procedures. Both the enzyme-linked immunosorbent assay and the quantitative immunofluorescence assay correlated well with serologically positive (indirect fluorescent-antibody titer greater than or e...

  17. Heterophilic antibodies interfering with radioimmunoassay. A false-positive pregnancy test

    International Nuclear Information System (INIS)

    A young woman with amenorrhea had a consistently positive pregnancy test result (serum radioimmunoassay measurement of #betta#-human chorionic gonadotropin hormone). No fetal or placental tissue was found after uterine curettage and exploratory laparotomy. The false-positive pregnancy test result was due to heterophilic antibovine and antigoat antibodies in the patient's serum. These antibodies interfered with radioimmunoassays using goat antibodies. This case shows that serum heterophilic antibodies can interfere with immunoassays and result in unnecessary diagnostic procedures and/or unnecessary treatment

  18. Development of Antibody-Coated Magnetite Nanoparticles for Biomarker Immobilization

    Directory of Open Access Journals (Sweden)

    Christian Chapa Gonzalez

    2014-01-01

    Full Text Available Magnetic nanoparticles (MNPs have great potential in biomedical applications because of their magnetic response offers the possibility to direct them to specific areas and target biological entities. Magnetic separation of biomolecules is one of the most important applications of MNPs because their versatility in detecting cancer biomarkers. However, the effectiveness of this method depends on many factors, including the type of functionalization onto MNPs. Therefore, in this study, magnetite nanoparticles have been developed in order to separate the 5′-nucleotidase enzyme (5eNT. The 5eNT is used as a bio-indicator for diagnosing diseases such as hepatic ischaemia, liver tumor, and hepatotoxic drugs damage. Magnetic nanoparticles were covered in a core/shell type with silica, aminosilane, and a double shell of silica-aminosilane. A ScFv (fragment antibody and anti-CD73 antibody were attached to the coated nanoparticles in order to separate the enzyme. The magnetic separation of this enzyme with fragment antibody was found to be 28% higher than anti-CD73 antibody and the enzyme adsorption was improved with the double shell due to the increased length of the polymeric chain. Magnetite nanoparticles with a double shell (silica-aminosilane were also found to be more sensitive than magnetite with a single shell in the detection of biomarkers.

  19. Development, Characterization and Application of Monoclonal Antibodies against Brazilian Dengue Virus Isolates

    OpenAIRE

    Camila Zanluca; Giovanny Augusto Camacho Antevere Mazzarotto; Juliano Bordignon; Claudia Nunes Duarte Dos Santos

    2014-01-01

    Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV) detection through the production and characterization of 22 monoclonal antibodies (mAbs) against Brazilian isolat...

  20. Development of a solid-phase radioimmunoassay for antibody to antigens of Babesia bovis infected bovine erythrocytes

    International Nuclear Information System (INIS)

    A quantitative solid-phase radioimmunoassay (RIA) has been developed for the purpose of ranking sera from exposed animals according to their titre of anti-B. bovis antibody. The antigen was a sonicate of lysed infected blood cells, and antibody binding was detected with 125I-labelled anti-bovine IgG. The assay was tested using sera from experimentally infected splenectomized and intact cattle and from animals resident in an endemic area. A high specificity for B. bovis was demonstrated. There was good agreement in identifying exposed cattle when the test was compared with an indirect fluorescent antibody (IFA) test although no correlation was seen between titres obtained in the two tests. Analysis of the effects of challenge infection with B. bovis on IFA and RIA titres in previously exposed animals illustrated that the RIA was a more sensitive test for detecting changes in antibody titre. (author)

  1. Development of a solid-phase radioimmunoassay for antibody to antigens of Babesia bovis infected bovine erythrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Kahl, L.P.; Anders, R.F.; Mitchell, G.F. (Walter and Eliza Hall Inst. of Medical Research, Parkville (Australia)); Callow, L.L.; Rodwell, B.J. (Animal Research Inst. Wacol (Australia). Tick Fever Research Centre)

    1982-06-01

    A quantitative solid-phase radioimmunoassay (RIA) has been developed for the purpose of ranking sera from exposed animals according to their titre of anti-B. bovis antibody. The antigen was a sonicate of lysed infected blood cells, and antibody binding was detected with /sup 125/I-labelled anti-bovine IgG. The assay was tested using sera from experimentally infected splenectomized and intact cattle and from animals resident in an endemic area. A high specificity for B. bovis was demonstrated. There was good agreement in identifying exposed cattle when the test was compared with an indirect fluorescent antibody (IFA) test although no correlation was seen between titres obtained in the two tests. Analysis of the effects of challenge infection with B. bovis on IFA and RIA titres in previously exposed animals illustrated that the RIA was a more sensitive test for detecting changes in antibody titre.

  2. Development and characterization of antibody reagents for detecting nanoparticles

    Science.gov (United States)

    Ravichandran, Supriya; Sullivan, Mark A.; Callahan, Linda M.; Bentley, Karen L.; Delouise, Lisa A.

    2015-11-01

    The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in solution. We present a proof-of-concept for the generation of NProbes and their use for detecting quantum dots and titanium dioxide NPs in vitro and in an ex vivo human skin model. Continued development and refinement of NProbes to detect NPs that vary in composition, shape, size, and surface coating will comprise a powerful tool kit that can be used to advance nanotechnology research particularly in the nanotoxicology and nanotherapeutics fields.The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in solution. We present a proof-of-concept for the generation of NProbes and their use for detecting quantum dots and titanium dioxide NPs in vitro and in an ex vivo human skin model. Continued development and refinement of NProbes to detect NPs that vary in composition, shape, size, and surface coating will comprise a powerful tool kit that can be used to advance nanotechnology research particularly in the nanotoxicology and nanotherapeutics fields. Electronic supplementary information (ESI) available: Figures and detailed methods of various techniques

  3. Longitudinal monitoring of the development of antifilarial antibodies and acquisition of Wuchereria bancrofti in a highly endemic area of Haiti.

    Directory of Open Access Journals (Sweden)

    Katy L Hamlin

    Full Text Available Antifilarial antibody testing has been established as a sensitive and specific method of diagnosing lymphatic filariasis. However, the development of serological responses to specific filarial antigens and their relationship to acquisition of infection is poorly understood. In order to evaluate whether the development of antigen specific antifilarial antibodies precedes microfilaremia and antigenemia, we compared the antibody responses of serum samples collected between 1990 and 1999 from a cohort of 142 Haitian children followed longitudinally. Antigen status was determined using the Og4C3 ELISA and the presence of microfilaremia was detected using microscopy. Antibody responses to Wb123, a Wuchereria bancrofti L3 antigen, were measured using a Luciferase Immunoprecipitation System (LIPS assay. Antibody responses to Bm14 and Bm33, Brugia malayi antigens and to a major surface protein (WSP from Wolbachia were analyzed using a multiplex bead assay. Over follow-up, 80 (56% of the children became antigen-positive and 30 (21% developed microfilaremia. Detectable antibody responses to Bm14, Bm33, Wb123, and WSP developed in 95%, 100%, 92%, and 29% of children, respectively. With the exception of WSP, the development of antibody responses generally preceded detection of filarial antigen. Our results show that antifilarial antibody responses can serve as an important epidemiological indicator in a sentinel population of young children and thus, may be valuable as tool for surveillance in the context of lymphatic filariasis elimination programs.

  4. Development of recombinant antigen array for simultaneous detection of viral antibodies.

    Directory of Open Access Journals (Sweden)

    Yi Liu

    Full Text Available Protein microarrays have been developed to study antibody reactivity against a large number of antigens, demonstrating extensive perspective for clinical application. We developed a viral antigen array by spotting four recombinant antigens and synthetic peptide, including glycoprotein G of herpes simplex virus (HSV type 1 and 2, phosphoprotein 150 of cytomegalovirus (CMV, Rubella virus (RV core plus glycoprotein E1 and E2 as well as a E1 peptide with the optimal concentrations on activated glass slides to simultaneously detect IgG and IgM against HSV1, HSV2, CMV and RV in clinical specimens of sera and cerebrospinal fluids (CSFs. The positive reference sera were initially used to measure the sensitivity and specificity of the array with the optimal conditions. Then clinical specimens of 144 sera and 93 CSFs were tested for IgG and IgM antibodies directed against HSV1, HSV2, CMV and RV by the antigen array. Specificity of the antigen array for viral antibodies detection was satisfying compared to commercial ELISA kits but sensitivity of the array varied relying on quality and antigenic epitopes of the spotting antigens. In short, the recombinant antigen array has potential to simultaneous detect multiple viral antibodies using minute amount (3 µl of samples, which holds the particularly advantage to detect viral antibodies in clinical CSFs being suspicious of neonatal meningitis and encephalitis.

  5. Detection of thrombocytic antibodies with the direct and indirect haemolysis inhibition test and the radioimmuno-Coombs test

    International Nuclear Information System (INIS)

    Methods of application of the direct and indirect haemolysis inhibition test were studied in order to optimise the test parameters: The ultimate aim was to standardize the test method and compare its sensitivity in detecting various platelet antibodies with platelet indirect radioactive Coombs-test and the platelet immunofluorescence test. (orig.)

  6. Evaluation Of Algorithms Of Anti- HIV Antibody Tests

    Directory of Open Access Journals (Sweden)

    Paranjape R.S

    1997-01-01

    Full Text Available Research question: Can alternate algorithms be used in place of conventional algorithm for epidemiological studies of HIV infection with less expenses? Objective: To compare the results of HIV sero- prevalence as determined by test algorithms combining three kits with conventional test algorithm. Study design: Cross â€" sectional. Participants: 282 truck drivers. Statistical analysis: Sensitivity and specificity analysis and predictive values. Results: Three different algorithms that do not include Western Blot (WB were compared with the conventional algorithm, in a truck driver population with 5.6% prevalence of HIV â€"I infection. Algorithms with one EIA (Genetic Systems or Biotest and a rapid test (immunocomb or with two EIAs showed 100% positive predictive value in relation to the conventional algorithm. Using an algorithm with EIA as screening test and a rapid test as a confirmatory test was 50 to 70% less expensive than the conventional algorithm per positive scrum sample. These algorithms obviate the interpretation of indeterminate results and also give differential diagnosis of HIV-2 infection. Alternate algorithms are ideally suited for community based control programme in developing countries. Application of these algorithms in population with low prevalence should also be studied in order to evaluate universal applicability.

  7. The study on the use of fragmented antibody for the development of therapeutic radiopharmaceutical

    International Nuclear Information System (INIS)

    This project was designed to develop the therapeutic radiopharmaceuticals for therapy and diagnosis of cancer using fragmented antibodies. The major activities to be carried out are as follows: - exploration of the key angiogenic factors involved in cancer, - development of radiolabeled-compounds using antibody fragments for minimal toxicity - In vitro/vivo investigation on the targeting ability of RI labeled antibody fragment

  8. Development of a novel monoclonal antibody with reactivity to a wide range of Venezuelan equine encephalitis virus strains

    Directory of Open Access Journals (Sweden)

    Phelps Amanda L

    2009-11-01

    Full Text Available Abstract Background There is currently a requirement for antiviral therapies capable of protecting against infection with Venezuelan equine encephalitis virus (VEEV, as a licensed vaccine is not available for general human use. Monoclonal antibodies are increasingly being developed as therapeutics and are potential treatments for VEEV as they have been shown to be protective in the mouse model of disease. However, to be truly effective, the antibody should recognise multiple strains of VEEV and broadly reactive monoclonal antibodies are rarely and only coincidentally isolated using classical hybridoma technology. Results In this work, methods were developed to reliably derive broadly reactive murine antibodies. A phage library was created that expressed single chain variable fragments (scFv isolated from mice immunised with multiple strains of VEEV. A broadly reactive scFv was identified and incorporated into a murine IgG2a framework. This novel antibody retained the broad reactivity exhibited by the scFv but did not possess virus neutralising activity. However, the antibody was still able to protect mice against VEEV disease induced by strain TrD when administered 24 h prior to challenge. Conclusion A monoclonal antibody possessing reactivity to a wide range of VEEV strains may be of benefit as a generic antiviral therapy. However, humanisation of the murine antibody will be required before it can be tested in humans. Crown Copyright © 2009

  9. Failure of a novel, rapid antigen and antibody combination test to detect antigen-positive HIV infection in African adults with early HIV infection.

    Directory of Open Access Journals (Sweden)

    William Kilembe

    Full Text Available BACKGROUND: Acute HIV infection (prior to antibody seroconversion represents a high-risk window for HIV transmission. Development of a test to detect acute infection at the point-of-care is urgent. METHODS: Volunteers enrolled in a prospective study of HIV incidence in four African cities, Kigali in Rwanda and Ndola, Kitwe and Lusaka in Zambia, were tested regularly for HIV by rapid antibody test and p24 antigen ELISA. Five subgroups of samples were also tested by the Determine Ag/Ab Combo test 1 Antigen positive, antibody negative (acute infection; 2 Antigen positive, antibody positive; 3 Antigen negative, antibody positive; 4 Antigen negative, antibody negative; and 5 Antigen false positive, antibody negative (HIV uninfected. A sixth group included serial dilutions from a p24 antigen-positive control sample. Combo test results were reported as antigen positive, antibody positive, or both. RESULTS: Of 34 group 1 samples with VL between 5x105 and >1.5x107 copies/mL (median 3.5x106, 1 (2.9% was detected by the Combo antigen component, 7 (20.6% others were positive by the Combo antibody component. No group 2 samples were antigen positive by the Combo test (0/18. Sensitivity of the Combo antigen test was therefore 1.9% (1/52, 95% CI 0.0, 9.9. One false positive Combo antibody result (1/30, 3.3% was observed in group 4. No false-positive Combo antigen results were observed. The Combo antigen test was positive in group 6 at concentrations of 80 pg/mL, faintly positive at 40 and 20 pg/mL, and negative thereafter. The p24 ELISA antigen test remained positive at 5 pg/mL. CONCLUSIONS: Although the antibody component of the Combo test detected antibodies to HIV earlier than the comparison antibody tests used, less than 2% of the cases of antigen-positive HIV infection were detected by the Combo antigen component. The development of a rapid point-of-care test to diagnose acute HIV infection remains an urgent goal.

  10. Multiplex method for initial complex testing of antibodies to blood transmitted diseases agents.

    Science.gov (United States)

    Poltavchenko, Alexander G; Nechitaylo, Oleg V; Filatov, Pavel V; Ersh, Anna V; Gureyev, Vadim N

    2016-10-01

    Initial screening of donors and population at high risk of infection with blood transmitted diseases involves a number of analyses using monospesific diagnostic systems, and therefore is expensive labor- and time-consuming process. The goal of this work is to construct a multiplex test enabling to carry out rapid initial complex testing at a low price. The paper describes a kit making it possible to detect simultaneously antibodies to six agents of the most significant blood transmitted diseases: HIV virus, hepatitis B and C viruses, cytomegalovirus, T. pallidum and T. gondii in blood products. The kit comprises multiplex dot-immunoassay based on plane protein arrays (immune chips) using colloidal gold conjugates and silver development. It provides an opportunity to carry out complex analysis within 70min at room temperature, and there is no need of well-qualified personnel. We compared laboratory findings of the kit with monospecific kits for ELISA produced by two Russian commercial companies. Dot-assay results correlate well with data obtained using commercial kits for ELISA. Furthermore, multiplex analysis is quicker and cheaper in comparison with ELISA and can be carried out in non-laboratory conditions. The kit for multiplex dot-immunoassay of antibodies to blood transmitted agents can significantly simplify initial complex testing. PMID:27497868

  11. Comparison of three ELISA methods with CLIF test for detection of Anti-dsDNA antibody

    OpenAIRE

    Feyza Çetin; Alparslan Toyran; Özlem Aytaç; Feride Alaca Coşkun; İpek Mumcuoğlu; Feyza Alp; Altan Aksoy

    2015-01-01

    INTRODUCTION[|]Systemic lupus erythematosus (SLE) is an autoimmune disease caused by antibodies which produced against antigens commonly on cell nuclei. According to the criteria of American College of Rheumatology (ACR), the immunological parameters which are used for diagnosis of SLE, are anti- nuclear antibodies (ANA) and anti- dsDNA autoantibodies. In this study it is aimed to investigate the specifity of three different ELISA tests by comparing with CLIF test, as ...

  12. Comparative evaluation of antibody detection tests to facilitate the diagnosis of multibacillary leprosy.

    Science.gov (United States)

    Duthie, Malcolm S; Orcullo, Florenda M; Abbelana, Junie; Maghanoy, Armi; Balagon, Marivic F

    2016-04-01

    Despite control efforts, leprosy persists as a significant health concern in many regions. Diagnosis is achieved by a combination of clinical, histopathological, and bacteriological examinations, each of which presents a barrier to expeditious diagnosis, particularly by non-experts. Immunological investigations in research laboratories have clearly indicated that antibody detection tests could aid the diagnosis of leprosy. In this study, we detected circulating antibodies with two rapid diagnostic tests (RDT) involving immunochromatographic lateral flow platforms and one rapid ELISA system. Leprosy patients were identified with a high degree of sensitivity in each assay (over 80 % in all; over 90 % among cases with bacterial indices >1+), although critical differences were observed in specificity. While the specificity of CTK OnSite Leprosy Ab Rapid Test and InBios Leprosy Detect™ fast ELISA were high (96.4 and 93.7 % in the general population, respectively), there was a marked reduction in OrangeLife NDO-LID® RDT (only 25.0 %). As anticipated, seropositivity rates were marginally higher in contacts of leprosy patients than in endemic controls. Although we observed a slight drop in test band intensity when blood, rather than serum, was used to develop OnSite Leprosy Ab Rapid Tests, the sensitivity and specificity of these tests was unaffected. When we contrasted test performance with clinical and bacteriological information, we found that RDT and ELISA results positively correlated with the bacteriological index. These data indicate that these assays could be a ready replacement of invasive, insensitive, and time consuming skin slit smear procedures that additionally require expert microscopic examinations. We propose that, due to their speed and point of care applicability, the RDT could be used as an initial entry point to the diagnostic protocols, with confirmation of results attained in a highly quantitative manner following serum transfer to a reference

  13. Development and Optimization of High-Throughput Methods To Measure Plasmodium falciparum-Specific Growth Inhibitory Antibodies

    OpenAIRE

    Persson, Kristina E. M.; Lee, Chee T.; Marsh, Kevin; Beeson, James G.

    2006-01-01

    Antibodies that inhibit replication of Plasmodium falciparum in erythrocytes are thought to be important both in acquired immunity to malaria and as mediators of immunity generated by candidate blood-stage vaccines. However, several constraints have limited the study of these functional antibodies in population studies and vaccine trials. We report the development and optimization of high-throughput growth inhibition assays with improved sensitivity that use minimal volumes of test serum. The...

  14. The development and specificity of antiidiotypic antibodies in renal transplant recipients receiving single-donor blood transfusions.

    Science.gov (United States)

    Phelan, D L; Rodey, G E; Anderson, C B

    1989-07-01

    Multiple pretransplant sera obtained from alloimmunized renal transplant recipients were tested for the presence of antiidiotypic-like antibodies (AB2) that inhibit donor-specific HLA antibodies in the microlymphocytotoxicity assay. Fourteen patients received repetitive single-donor blood transfusions (SDT). In this patient group, sera were collected prior to each blood transfusion and prior to transplantation. Three additional patients were studied in whom prior donor-specific HLA antibodies had been lost over a period of 6 months preceding transplantation. Donor-specific AB2-like antibodies were found in the sera of 13/14 SDT patients who did not develop HLA antibodies, and in the 3 patients who had lost donor-specific HLA antibodies. All patients had received prior random blood transfusions in the year preceding the study. Five (38%) of the SDT patients had detectable donor-specific AB2 prior to the initiation of single-donor blood transfusion, presumably related to previous blood transfusions. In the remaining six SDT patients in whom complete serum sets were available, AB2 always appeared after the first blood transfusion. The specificity of HLA antibodies inhibited by AB2 was studied, and antibodies against HLA-A, -B, -C, -DR, and DQw were all identified. Thus, there was no predilection for patients to develop AB2 against locus-specific HLA gene products. This study also confirms the apparent polymorphism of putative crossreactive idiotypes. Approximately 25% of donor-specific HLA antibodies were not inhibited by relevant AB2. This study confirms and extends previous observations that alloimmunization is associated in many patients with the development of antiidiotypic-like antibodies that are capable of inhibiting the binding and cytotoxicity of HLA alloantibodies. PMID:2473550

  15. Clinical factors associated with a Candida albicans Germ Tube Antibody positive test in Intensive Care Unit patients

    OpenAIRE

    Martín-Mazuelos Estrella; Giménez María J; Ramírez Paula; Camarena Juan J; Cuétara María S; Alkorta Miriam; Quindós Guillermo; Zaragoza Rafael; Pemán Javier; Linares-Sicilia María J; Pontón José

    2011-01-01

    Abstract Background Poor outcomes of invasive candidiasis (IC) are associated with the difficulty in establishing the microbiological diagnosis at an early stage. New scores and laboratory tests have been developed in order to make an early therapeutic intervention in an attempt to reduce the high mortality associated with invasive fungal infections. Candida albicans IFA IgG has been recently commercialized for germ tube antibody detection (CAGTA). This test provides a rapid and simple diagno...

  16. Radiometric cytolysis inhibition assay, a new rapid test for neutralizing antibodies to intact and trypsin-cleaved poliovirus

    International Nuclear Information System (INIS)

    We have developed a new rapid test, the radiometric cytolysis inhibition assay (RACINA), for the determination of neutralizing poliovirus antibodies. HeLa cells prelabeled with 51Cr, [3H]leucine, or, preferentially, with [3H]uridine are used as sensitive quantitative indicators of residual infectious virus. Both suspensions and monolayer cultures of the indicator cells can be used. Neutralization of a fraction of a high-titer virus preparation can be scored after the first replication cycle at 8 to 10 h. By lowering the incubation temperature to 30 degree C, the completion of the cytolysis due to the first replication cycle of poliovirus was delayed beyond 21 h. This makes it possible to use the RACINA, unlike the standard microneutralization assay, for measuring antibodies to trypsin-cleaved polioviruses. The RACINA was found to be as sensitive as and more reproducible than the standard microneutralization assay in the measurement of neutralizing poliovirus antibodies. The RACINA is a rapid and reliable test for neutralizing antibodies and in principle it may be applicable for quantitation of neutralizing antibodies to other cytolytic agents as well

  17. AUTOMATED TESTING IN DEVELOPMENT PHASE

    Directory of Open Access Journals (Sweden)

    SUNIL L. BANGARE

    2012-02-01

    Full Text Available In software development the applications are tested in testing phase of software development process. So testing of application is not possible without complete development of module/application. It takes additional time in completion of software development. So this paper proposed the model for tool which provides the way to developer to test his code/application in development phase itself. The model also helps in java API (application programmable interface testing. With this tool, developer can able to test his code/module automaticallyconsidering all the aspect of testing. In this approach predefined test cases are loaded for testing, and thousands of test cases are run at same time and application is tested by developer. So it helps in regression testing. Hence it helps in reducing software development period. Ultimately it saves the people resources, hardware and software resources.

  18. Commercial serological antibody detection tests for the diagnosis of pulmonary tuberculosis: a systematic review.

    Directory of Open Access Journals (Sweden)

    Karen R Steingart

    2007-06-01

    Full Text Available BACKGROUND: The global tuberculosis epidemic results in nearly 2 million deaths and 9 million new cases of the disease a year. The vast majority of tuberculosis patients live in developing countries, where the diagnosis of tuberculosis relies on the identification of acid-fast bacilli on unprocessed sputum smears using conventional light microscopy. Microscopy has high specificity in tuberculosis-endemic countries, but modest sensitivity which varies among laboratories (range 20% to 80%. Moreover, the sensitivity is poor for paucibacillary disease (e.g., pediatric and HIV-associated tuberculosis. Thus, the development of rapid and accurate new diagnostic tools is imperative. Immune-based tests are potentially suitable for use in low-income countries as some test formats can be performed at the point of care without laboratory equipment. Currently, dozens of distinct commercial antibody detection tests are sold in developing countries. The question is "do they work?" METHODS AND FINDINGS: We conducted a systematic review to assess the accuracy of commercial antibody detection tests for the diagnosis of pulmonary tuberculosis. Studies from all countries using culture and/or microscopy smear for confirmation of pulmonary tuberculosis were eligible. Studies with fewer than 50 participants (25 patients and 25 control participants were excluded. In a comprehensive search, we identified 68 studies. The results demonstrate that (1 overall, commercial tests vary widely in performance; (2 sensitivity is higher in smear-positive than smear-negative samples; (3 in studies of smear-positive patients, Anda-TB IgG by enzyme-linked immunosorbent assay shows limited sensitivity (range 63% to 85% and inconsistent specificity (range 73% to 100%; (4 specificity is higher in healthy volunteers than in patients in whom tuberculosis disease is initially suspected and subsequently ruled out; and (5 there are insufficient data to determine the accuracy of most

  19. A pathologist-in-the-loop IHC antibody test selection using the entropy-based probabilistic method

    Directory of Open Access Journals (Sweden)

    Dmitriy Shin

    2012-01-01

    Full Text Available Background: Immunohistochemistry (IHC is an important tool to identify and quantify expression of certain proteins (antigens to gain insights into the molecular processes in a diseased tissue. However, it is a challenge for pathologists to remember the discriminative characteristics of the growing number of such antigens across multiple diseases. The complexity of their expression patterns, fueled by continuous discoveries in molecular pathology, gives rise to a combinatorial explosion that places an unprecedented burden on a practicing pathologist and therefore increases cost and variability of IHC studies. Materials and Methods: To tackle these issues, we have developed antibody test optimized selection method, a novel informatics tool to help pathologists in improving the IHC antibody selection process. The method uses extensions of Shannon′s information entropies and Bayesian probabilities to dynamically build an efficient diagnostic tree. Results: A comparative analysis of our method with the expert and World Health Organization classification guidelines showed that the proposed method brings threefold reduction in number of antibody tests required to reach a diagnostic conclusion. Conclusion: The developed method can significantly streamline the antibody test selection process, decrease associated costs and reduce inter- and intrapathologist variability in IHC decision-making.

  20. Diagnosis and Clinical Virology of Lassa Fever as Evaluated by Enzyme-Linked Immunosorbent Assay, Indirect Fluorescent-Antibody Test, and Virus Isolation

    OpenAIRE

    Bausch, D. G.; Rollin, P E; Demby, A H; Coulibaly, M.; Kanu, J.; Conteh, A. S.; Wagoner, K. D.; McMullan, L. K.; Bowen, M. D.; Peters, C. J.; Ksiazek, T. G.

    2000-01-01

    The Lassa virus (an arenavirus) is found in West Africa, where it sometimes causes a severe hemorrhagic illness called Lassa fever. Laboratory diagnosis has traditionally been by the indirect fluorescent-antibody (IFA) test. However, enzyme-linked immunosorbent assays (ELISAs) for Lassa virus antigen and immunoglobulin M (IgM) and G (IgG) antibodies have been developed that are thought to be more sensitive and specific. We compared ELISA and IFA testing on sera from 305 suspected cases of Las...

  1. Validation of pre-coated ELISA tests to detect antibodies against T. congolense and T. vivax

    International Nuclear Information System (INIS)

    The anti-trypanosomal antibody detecting enzyme linked immunosorbent assay (ELISA) was first described in 1977 and was further developed for use in large scale surveys in Zimbabwe. More recently, the IAEA initiated a programme to improve the robustness and standardisation of the assay. The IAEA supplied plates pre-coated with either a crude T. congolense or T. vivax antigen and the reagents necessary for analysing samples. Parasitologically positive and negative sera were used to validate and determine the cut-off values of the two tests. The samples were tested and results analysed using a variety of cut-off values. The tests provided similar information although the T. congolense pre-coated plates gave significantly higher optical density values than the plates coated with T. vivax. Sensitivity and specificity values were calculated using the different cut-off points. Results indicate that the test using T. congolense antigen had the highest specificity and sensitivity for a given cut-off value. Although the test could distinguish positive from negative sera, it was quite difficult to provide a suitable cut-off value, but the value should be dictated by the use of the test. (author)

  2. Hepatitis B core antibody testing in Indian blood donors: A double-edged sword!

    Directory of Open Access Journals (Sweden)

    R N Makroo

    2012-01-01

    Full Text Available Background: Until lately, anti-HBc antibodies were considered an effective marker for occult Hepatitis B virus (HBV infection and have served their role in improving blood safety. But, with the development of advanced tests for HBV DNA detection, the role of anti-HBc in this regard stands uncertain. Materials and Methods: Anti-HBc and HBsAg ELISA and ID-NAT tests were run in parallel on donor blood samples between April 1, 2006 and December 31, 2010 at the Department of Transfusion Medicine, Indraprastha Apollo Hospitals, New Delhi. A positive ID-NAT was followed by Discriminatory NAT assay. Results: A total of 94 247 samples were tested with a total core positivity rate of 10.22%. We identified nearly 9.17% of donors who were reactive for anti-HBc and negative for HBsAg and HBV DNA. These are the donors who are potentially non-infectious and may be returned to the donor pool. Conclusion: Although anti HBc testing has a definite role in improving blood safety, centers that have incorporated NAT testing may not derive any additional benefit by performing anti-HBc testing, especially in resource-limited countries like ours.

  3. Heterogeneity of laboratory test results for antiphospholipid antibodies in patients treated with chlorpromazine and other phenothiazines.

    Science.gov (United States)

    Lillicrap, D P; Pinto, M; Benford, K; Ford, P M; Ford, S

    1990-06-01

    Ninety-seven psychiatric patients who have been treated with the antipsychotic drug chlorpromazine or another phenothiazine have been investigated for the presence of antiphospholipid antibodies. A variety of coagulation studies and specific antiphospholipid immunoassays were performed to define the spectrum of antigen specificity of these antibodies. Coagulation studies showed an increasing sensitivity for the lupus anticoagulant with reagents of differing phospholipid content. Prolonged activated partial thromboplastin times (APTTs) were found in five patients with the use of an insensitive APTT reagent and in 14 patients with a lower phospholipid content reagent. In every case, attempted correction of the clotting time with normal plasma was unsuccessful. Twenty-one patients had abnormal kaolin clotting time profiles. In seven of these patients, test results with both APTT reagents had been normal. Antibody reactivity was tested against three negatively charged phospholipids, phosphatidyl-serine, cardiolipin, and phosphatidylinositol. Only five patients demonstrated reactivity against phosphatidylinositol, whereas high antibody titers were observed in 28 patients against one or both of phosphatidylserine and cardiolipin. Twenty-three of these patients were found to have elevated anticardiolipin-specific IgM antibodies. Overall, 41 of the patients had at least one laboratory abnormality suggestive of antiphospholipid antibody activity. Seven of the 26 patients, taking phenothiazines other than chlorpromazine, had positive test results for antiphospholipid antibodies. No clinical thromboembolic events were recorded in any patient. These findings demonstrate the heterogeneity of antiphospholipid antibody specificity induced in patients treated with various phenothiazine drugs and indicate that none of these patterns of reactivity marks a predisposition for thromboembolism in this population. PMID:1971739

  4. Challenges in Antibody Development against Tn and Sialyl-Tn Antigens

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    Liliana R. Loureiro

    2015-08-01

    Full Text Available The carbohydrate antigens Tn and sialyl-Tn (STn are expressed in most carcinomas and usually absent in healthy tissues. These antigens have been correlated with cancer progression and poor prognosis, and associated with immunosuppressive microenvironment. Presently they are used in clinical trials as therapeutic vaccination, but with limited success due to their low immunogenicity. Alternatively, anti-Tn and/or STn antibodies may be used to harness the immune system against tumor cells. Whilst the development of antibodies against these antigens had a boost two decades ago for diagnostic use, so far no such antibody entered into clinical trials. Possible limitations are the low specificity and efficiency of existing antibodies and that novel antibodies are still necessary. The vast array of methodologies available today will allow rapid antibody development and novel formats. Following the advent of hybridoma technology, the immortalization of human B cells became a methodology to obtain human monoclonal antibodies with better specificity. Advances in molecular biology including phage display technology for high throughput screening, transgenic mice and more recently molecularly engineered antibodies enhanced the field of antibody production. The development of novel antibodies against Tn and STn taking advantage of innovative technologies and engineering techniques may result in innovative therapeutic antibodies for cancer treatment.

  5. THE INVESTIGATION OF BRUCELLA ANTIBODY WITH MILK RING TEST AND AGGLUTINATION TEST IN MILK COLLECTED FROM SAMSUN REGION

    Directory of Open Access Journals (Sweden)

    Goknur TERZI

    2006-06-01

    Full Text Available In this study Brucella antibodies were investigated with agglutination test (Whey-AT and Milk Ring Test (MRT in a total of 100 milk samples as 50 of cow milk and 50 of goat milk collected from center and villages of Samsun. According to MRT Brucella antibodies was positive at 10 samples (20 % of cow milk and 6 samples (12 % of goat milk. In cow milk, 4 (8 % positive, 3 (6 % suspicious and 43 (86 % negative samples; in goat milk 3 (6 % positive, 2 (4 % suspicious and 45 (90 % negative samples were determined according to antibodies titre of serum agglutination test (Whey-AT. [TAF Prev Med Bull 2006; 5(3.000: 196-203

  6. Comparative efficacy of antigen and antibody detection tests for human trichinellosis

    International Nuclear Information System (INIS)

    Sera collected from patients with suspected or confirmed exposure to Trichinella spiralis were tested for circulating parasite antigens and antiparasite antibodies. Using an immunoradiometric assay, excretory--secretory antigens from muscle-stage larvae of T. spiralis were detected in the sera of 47% of 62 patients with clinical trichinellosis and 13% of 39 patients without clinical signs but suspected of exposure to infected meat. In comparison, antibodies were detected using an indirect immunofluorescent test in the circulation of 100% of the 62 patients with clinical trichinellosis and 46% of the 39 patients with suspected exposure. The presence of antibodies specific to excretory-secretory products of T. spiralis muscle larvae was confirmed in the majority of the samples tested by a monoclonal antibody-based competitive inhibition assay. These results indicate that antibody detection is a more sensitive diagnostic method for human trichinellosis, but that antigen detection might be a useful confirmatory test because it is a direct demonstration of parasite products in the circulation

  7. Comparative Evaluation of Seven Commercial Tests for Detection of Heterophile Antibody in Infectious Mononucleosis

    OpenAIRE

    Skulnick, Martin; Low, Donald E.; Simor, Andrew E.; Patel, Mohan; George, Pauline; Chua, Robert

    1992-01-01

    Detection of heterophile antibodies in infectious mononucleosis is the most rapid and cost-effective method for confirming the clinical diagnosis of the disease. This study compared seven commercial test kits (the Oxoid Infectious Mononucleosis Kit [Oxoid Ltd], Immunoscan Im-Latex [Baxter Travenol], Mono-Latex [Wampole Laboratories], Monospot and Im Screen Test [Ortho Diagnostics], Immunoscan Im-RBC Test [Baxter Travenol], and Infectious Mononucleosis Test [NCS Diagnostics]) to the Davidsohn ...

  8. Development of Biodegradable Nanocarriers Loaded with a Monoclonal Antibody

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    Andrew Gdowski

    2015-02-01

    Full Text Available Treatments utilizing monoclonal antibody therapeutics against intracellular protein-protein interactions in cancer cells have been hampered by several factors, including poor intracellular uptake and rapid lysosomal degradation. Our current work examines the feasibility of encapsulating monoclonal antibodies within poly(lactic-co-glycolic acid (PLGA nanoparticles using a water/oil/water double emulsion solvent evaporation technique. This method can be used to prepare protective polymeric nanoparticles for transporting functional antibodies to the cytoplasmic compartment of cancer cells. Nanoparticles were formulated and then characterized using a number of physical and biological parameters. The average nanoparticle size ranged from 221 to 252 nm with a low polydispersity index. Encapsulation efficiency of 16%–22% and antibody loading of 0.3%–1.12% were observed. The antibody molecules were released from the nanoparticles in a sustained manner and upon release maintained functionality. Our studies achieved successful formulation of antibody loaded polymeric nanoparticles, thus indicating that a PLGA-based antibody nanoformulation is a promising intracellular delivery vehicle for a large number of new intracellular antibody targets in cancer cells.

  9. Development of Robust and Standardized Cantilever Sensors Based on Biotin/Neutravidin Coupling for Antibody Detection

    Directory of Open Access Journals (Sweden)

    Christoph Gerber

    2013-04-01

    Full Text Available A cantilever-based protein biosensor has been developed providing a customizable multilayer platform for the detection of antibodies. It consists of a biotin-terminated PEG layer pre-functionalized on the gold-coated cantilever surface, onto which NeutrAvidin is adsorbed through biotin/NeutrAvidin specific binding. NeutrAvidin is used as a bridge layer between the biotin-coated surface and the biotinylated biomolecules, such as biotinylated bovine serum albumin (biotinylated BSA, forming a multilayer sensor for direct antibody capture. The cantilever biosensor has been successfully applied to the detection of mouse anti-BSA (m-IgG and sheep anti-BSA(s-IgG antibodies. As expected, the average differential surface stress signals of about 5.7 ± 0.8 ´ 10−3 N/m are very similar for BSA/m-IgG and BSA/s-IgG binding, i.e., they are independent of the origin of the antibody. A statistic evaluation of 112 response curves confirms that the multilayer protein cantilever biosensor shows high reproducibility. As a control test, a biotinylated maltose binding protein was used for detecting specificity of IgG, the result shows a signal of bBSA layer in response to antibody is 5.8 ´ 10−3 N/m compared to bMBP. The pre-functionalized biotin/PEG cantilever surface is found to show a long shelf-life of at least 40 days and retains its responsivity of above 70% of the signal when stored in PBS buffer at 4 °C. The protein cantilever biosensor represents a rapid, label-free, sensitive and reliable detection technique for a real-time protein assay.

  10. Development of Robust and Standardized Cantilever Sensors Based on Biotin/Neutravidin Coupling for Antibody Detection

    Science.gov (United States)

    Zhang, Jiayun; Lang, Hans Peter; Battiston, Felice; Backmann, Natalija; Huber, Francois; Gerber, Christoph

    2013-01-01

    A cantilever-based protein biosensor has been developed providing a customizable multilayer platform for the detection of antibodies. It consists of a biotin-terminated PEG layer pre-functionalized on the gold-coated cantilever surface, onto which NeutrAvidin is adsorbed through biotin/NeutrAvidin specific binding. NeutrAvidin is used as a bridge layer between the biotin-coated surface and the biotinylated biomolecules, such as biotinylated bovine serum albumin (biotinylated BSA), forming a multilayer sensor for direct antibody capture. The cantilever biosensor has been successfully applied to the detection of mouse anti-BSA (m-IgG) and sheep anti-BSA(s-IgG) antibodies. As expected, the average differential surface stress signals of about 5.7 ± 0.8 × 10−3 N/m are very similar for BSA/m-IgG and BSA/s-IgG binding, i.e., they are independent of the origin of the antibody. A statistic evaluation of 112 response curves confirms that the multilayer protein cantilever biosensor shows high reproducibility. As a control test, a biotinylated maltose binding protein was used for detecting specificity of IgG, the result shows a signal of bBSA layer in response to antibody is 5.8 × 10−3 N/m compared to bMBP. The pre-functionalized biotin/PEG cantilever surface is found to show a long shelf-life of at least 40 days and retains its responsivity of above 70% of the signal when stored in PBS buffer at 4 °C. The protein cantilever biosensor represents a rapid, label-free, sensitive and reliable detection technique for a real-time protein assay. PMID:23604028

  11. THE INVESTIGATION OF BRUCELLA ANTIBODY WITH MILK RING TEST AND AGGLUTINATION TEST IN MILK COLLECTED FROM SAMSUN REGION

    OpenAIRE

    Goknur TERZI

    2006-01-01

    In this study Brucella antibodies were investigated with agglutination test (Whey-AT) and Milk Ring Test (MRT) in a total of 100 milk samples as 50 of cow milk and 50 of goat milk collected from center and villages of Samsun. According to MRT Brucella antibodies was positive at 10 samples (20 %) of cow milk and 6 samples (12 %) of goat milk. In cow milk, 4 (8 %) positive, 3 (6 %) suspicious and 43 (86 %) negative samples; in goat milk 3 (6 %) positive, 2 (4 %) suspicious and 45 (90 %) negativ...

  12. Development of a sensitive and specific epitope-blocking ELISA for universal detection of antibodies to human enterovirus 71 strains.

    Directory of Open Access Journals (Sweden)

    Fang He

    Full Text Available BACKGROUND: Human Enterovirus 71 (EV71 is a common cause of hand, foot and mouth disease (HFMD in young children. It is often associated with severe neurological diseases and mortalities in recent outbreaks across the Asia Pacific region. Currently, there is no efficient universal antibody test available to detect EV71 infections. METHODOLOGY/PRINCIPAL FINDING: In the present study, an epitope-blocking ELISA was developed to detect specific antibodies to human EV71 viruses in human or animal sera. The assay relies on a novel monoclonal antibody (Mab 1C6 that specifically binds to capsid proteins in whole EV71 viruses without any cross reaction to any EV71 capsid protein expressed alone. The sensitivity and specificity of the epitope-blocking ELISA for EV71 was evaluated and compared to microneutralization using immunized animal sera to multiple virus genotypes of EV71 and coxsackieviruses. Further, 200 serum sample from human individuals who were potentially infected with EV71 viruses were tested in both the blocking ELISA and microneutralization. Results indicated that antibodies to EV71 were readily detected in immunized animals or human sera by the epitope blocking ELISA whereas specimens with antibodies to other enteroviruses yielded negative results. This assay is not only simpler to perform but also shows higher sensitivity and specificity as compared to microneutralization. CONCLUSION: The epitope-blocking ELISA based on a unique Mab 1C6 provided highly sensitive and 100% specific detection of antibodies to human EV71 viruses in human sera.

  13. Immunogenicity screening assay development for a novel human-mouse chimeric anti-CD147 monoclonal antibody (Metuzumab).

    Science.gov (United States)

    Mi, Li; Li, Wei; Li, Maohua; Chen, Tao; Wang, Muyang; Sun, Le; Chen, Zhinan

    2016-06-01

    The clinical effect of patient immune responses to therapeutic antibodies affect product safety and efficacy, which makes the development of valid, sensitive immune assays a key aspect of antibody drug development. In this paper, we reported the generations of mouse monoclonal and Cynomolgus monkey polyclonal antibodies against the anti-CD147 antibody (Metuzumab) as the internal standards and the positive controls. Seven mouse monoclonal antibodies were shown to recognize both (Fab)2 and full length of Metuzumab, but not the control normal human IgGs, and monoclonal anti-Metuzumab, Clone 2D9 was chosen to be used as the internal standard for anti-Metuzumab study. A Bridging ELISA assay was developed by coating the wells with the antibody drug, and the anti-drug antibody (ADA) in the animal sera were detected by enzyme-labeled antibody. Its limit of detection (LOD) was determined to be 0.39ng/ml of anti-Metuzumab antibody (ADA) with linear range between 0.39-50ng/ml and R(2)=0.994. For normal monkey sera, a minimal dilution was determined to be 1:80. However, very different from peptide or other protein drugs, strong interferences from the residual antibody drugs were observed from most of the testing monkey sera in the preclinical study. It was experimentally determined that the concentration of the residual antibody drug in the assay have to be lower than 1μg/ml, so the assays were carried out at 1:100 dilution of the monkey sera. In the pre-clinical study, 32 monkeys were treated with escalating doses of Metuzumab between 0, 10, 50, 200mg/kg for 13 times over 13weeks of time period. 16 of them were terminated right after the last injection, while the other 16 were rested for additional 4weeks before termination. Afraid to miss any positive response to antibody drug, sera samples were collected at six time points, including 2-, 6- and 10-weeks post 1st dose, prior to last dose, and 2-, 4-weeks into recovery. The highest positive rates were seen with the Medium

  14. A multicenter evaluation of a new antibody test kit for lymphatic filariasis employing recombinant Brugia malayi antigen Bm-14

    OpenAIRE

    Weil, Gary J; Curtis, Kurt C.; Fischer, Peter U.; Kimberly Y Won; Lammie, Patrick J; Joseph, Hayley; Melrose, Wayne D; Brattig, Norbert W.

    2010-01-01

    Antibody tests are useful for mapping the distribution of lymphatic filariasis (LF) in countries and regions and for monitoring progress in elimination programs based on mass drug administration (MDA). Prior antibody tests have suffered from poor sensitivity and/or specificity or from a lack of standardization. We conducted a multicenter evaluation of a new commercial ELISA that detects IgG4 antibodies to the recombinant filarial antigen Bm14. Four laboratories tested a shared panel of coded ...

  15. Pseudo-plaque reduction neutralization test (PPRNT for the measurement of neutralizing antibodies to Crimean-Congo hemorrhagic fever virus

    Directory of Open Access Journals (Sweden)

    Canakoglu Nurettin

    2013-01-01

    Full Text Available Abstract Background Crimean-Congo hemorrhagic fever virus (CCHFV is a tick-borne virus of the genus Nairovirus family Bunyaviridae, which are enveloped viruses containing tripartite, negative polarity, single-stranded RNA. CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Europe. Currently, there are no specific treatments or licensed vaccines available for CCHFV. Recently, two research groups have found adult mice with defective interferon responses allowed to lethal CCHFV infection. These mouse models could provide invaluable information for further studies. Efforts to develop a vaccine against CCHFV are being made. To determine the efficacy of vaccine candidates it is important to conduct serological studies that can accurately measure levels of protective antibodies. In the present study, a pseudo-plaque reduction neutralization test (PPRNT based on enzyme-catalyzed color development of infected cells probed with anti-CCHFV antibodies was used to measure neutralization antibody of CCHFV. Methods Sixty-nine human serum samples (20 acute and 49 convalescent were tested. The presence of CCHFV antibodies was determined and confirmed by a commercial ELISA kit. CCHFV RNA was determined by RT-PCR. All the samples were analyzed by PPRNT and fluorescent focus reduction neutralization test (FFRNT to measure of CCHFV-neutralizing antibodies. Results Pseudo-plaque reduction neutralization test showed a high sensitivity (98%, specificity (100% and agreement (96,6% in qualitative comparison with those of the FFRNT. There was a high correlation between the titers obtained in PPRNT and FFRNT (R2 = 0.92. The inter- and intra-assay variation of PPRNT revealed good reproducibility and positive cut-off of PPRNT was defined as 1:4 by the geometric mean titers for the individual samples distributed. Conclusion The pseudo-plaque reduction neutralization test described in this study is a fast, reproducible and sensitive

  16. Accuracy of diagnostic antibody tests for coeliac disease in children

    DEFF Research Database (Denmark)

    Giersiepen, Klaus; Lelgemann, Monika; Stuhldreher, Nina; Ronfani, Luca; Husby, Steffen; Koletzko, Sibylle; Korponay-Szabó, Ilma R

    2012-01-01

    The aim of this study was to summarise the evidence from 2004 to September 2009 on the performance of laboratory-based serological and point of care (POC) tests for diagnosing coeliac disease (CD) in children using histology as reference standard....

  17. Developments of sensitive immunoassays for detection of antibodies against hepatitis B surface antigen

    International Nuclear Information System (INIS)

    Three micro solid phase immunoassays (a micro-SPRIA and two ELISA techniques) were developed and tested for the detection of anti-HBs antibodies. Two different crosslinkers (glutaraldehyde and N-succinimidyl 3-(2-pyridyldithio) propionate) were used to couple a goat anti-mouse IgG reagent to alkaline phosphatase for use as enzyme-labeled probes in the two ELISA tests. With the latter cross-linker, a defined conjugate with a 1 : 1 antibody-enzyme molar ratio was obtained. The sensitivities of micro-SPRIA and the two types of ELISA were compared to that of the commercial solid phase radioimmunoassay AUSAB test. All three microtests were significantly more sensitive than the AUSAB test. The ELISA using the glutaraldehyde cross-linked conjugate was 3-5 times less sensitive than micro-SPRIA, while the ELISA using the disulfide-linked conjugate was 2.6-4.0 times more sensitive than micro-SPRIA. (Auth.)

  18. Interlaboratory testing of porcine sera for antibodies to porcine circovirus type 2

    DEFF Research Database (Denmark)

    McNair, I.; Marshall, M.; McNeilly, F.; Bøtner, Anette; Ladekjær-Mikkelsen, A.S.; Vincent, I.; Herrmann, Brigitte; Sanchez, R.; Rhodes, C.

    2004-01-01

    A panel of 20 porcine sera was distributed to 5 laboratories across Europe and Canada. Each center was requested to test the sera for the presence of porcine circovirus type 2 antibodies using the routine assays, indirect immunofluorescence assay (IFA) and indirect immunoperoxidase monolayer assay...

  19. Laboratory evaluation of three rapid diagnostic tests for dual detection of HIV and Treponema pallidum antibodies.

    Science.gov (United States)

    Humphries, Romney M; Woo, Jennifer S; Chung, Jun Ho; Sokovic, Anita; Bristow, Claire C; Klausner, Jeffrey D

    2014-12-01

    The performance of three research-use-only, dual HIV and syphilis rapid diagnostic tests (RDTs) was evaluated for 150 patient serum samples and compared to reference HIV and Treponema pallidum antibody detection methods. The RDTs performed comparably, with sensitivities of 93 to 99% and specificities of 97 to 100%. The kappa statistic between the RDTs was 0.95. PMID:25297332

  20. The Factor Structure of The HIV Antibody Testing Attitude Scale in Four African Countries

    OpenAIRE

    K. Peltzer; Mpofu, E

    2013-01-01

    Objective: To determine the facture structure of the HIVAntibody Testing Attitude Scale (HTAS) in an Africanpopulation.Method: 760 first-year African university students fromNigeria, South Africa, Uganda and Zimbabwe weresurveyed using the HIV Antibody Testing Attitude Scale.Factor structure was determined by using the principalcomponent analysis with varimax rotation.Results: Five components accounting for 51% of thetotal variance were identified. The first factor(eigenvalue: 5.11) accounted...

  1. PCR and antibody methods: Research compares two cattle feed tests that detect bovine byproduct contaminants

    OpenAIRE

    Sawyer, Mary M.; Smith, Wayne L.; Rensen, Gabriel J.; Osburn, Bennie I.; Cullor, James S.

    2005-01-01

    Preventing the spread of mad cow disease through contaminated cattle feed is a major concern of beef and dairy producers, regulators and consumers around the world. Routine testing of cattle feeds for the presence of banned substances is a critical control point in assuring animal health and food safety. We compared the results of two test procedures (a real-time polymerase chain reaction [PCR] assay and a commercially available ruminant antibody detection kit) on five cattle rations spiked w...

  2. Development of radiolabelling techniques of anti-CEA monoclonal antibody

    International Nuclear Information System (INIS)

    The purpose of this work was to label monoclonal and polyclonal antibodies with 99Tcm such as the ior-CEA-1 antibody and polyclonal IgG using a direct method, to check the radiochemical and biological behavior of labelled products, to prepare it under sterile and apyrogenic conditions as a lyophilized kit and to employ it in clinical trials. In addition, a photoactivation method was used to label polyclonal IgG with 99Tcm and to compare with the established method using mercaptoethanol (2-ME) as the reducing agent. Finally polyclonal IgG was labelled using an indirect method in which a chelator was covalently attached to the protein and the 99Tcm added as glucoheptonate complex. The properties of 99Tcm when labelled with monoclonal and polyclonal antibodies by different methods were assessed by in vitro and in vivo studies

  3. Seroprevalence of Mycoplasma gallisepticum antibody by ELISA and serum plate agglutination test of laying chicken

    Directory of Open Access Journals (Sweden)

    Md. Zulfekar Ali

    2015-01-01

    Full Text Available Aim: Mycoplasma gallisepticum (MG is important avian pathogen responsible for chronic respiratory disease of chicken and turkeys, which result in large economic loss for the poultry industry. The objectives of this study were determination of seroprevalence of MG antibody of commercial layer chicken at laying period in selected areas of Bangladesh. Materials and Methods: A total of 563 blood samples were collected randomly from selected commercial layer chickens at laying period during the period from July to December, 2013. Indirect enzyme linked immunosorbent assay (iELISA and serum plate agglutination (SPA test were performed to detect the presence of antibodies against MG. Results: Of 563 samples, 64.47% and 56.13% showed an overall prevalence of MG antibodies in iELISA and SPA test respectively. Prevalence of MG was recorded the highest (69.63% at 50-55 weeks of age compared with lowest (53.26% at 56-61 weeks of age (p<0.05. Significant (p<0.05 effect of breed were observed in the seroprevalence of MG infection in layer birds in the present study. The overall, 68.77%, 63.74% and 59.37% prevalence were found respectively in sonali, ISA Brown and White leg horn. The prevalence of MG antibodies was the highest (70.13% in December followed by November (68%, October (65.67%, August (63.46%, September (58.54% and July (51.78% month. The seroprevalence of MG antibodies was higher (69.63% in most of the large flocks and lower (56.82% in small flocks. Conclusion: Therefore, might be suggested that the commercial layer farms should be routinely checked to monitor MG infection and the reactor birds should be culled since MG organism has the potential to transmit vertically. The correlation between MG antibody in month and flock size was not significant (p=0.359 and p=0.868, respectively.

  4. Reliability of urinary tests for antibody to Helicobacter pylori in patients with pulmonary tuberculosis

    Institute of Scientific and Technical Information of China (English)

    Takatsugu Yamamoto; Taro Ishii; Tomotaka Kawakami; Yoko Sase; Chiaki Horikawa; Nozomu Aoki; Masaki Sanaka; Yasushi Kuvama

    2005-01-01

    AIM: Although the quality of currently available urinary tests for detecting antibody to Helicobacter pylori (H pylori)have been proved in some populations, the accuracy has not been studied regarding patients who suffer from pulmonary tuberculosis with multi-drug treatments. The present study was conducted to evaluate the accuracy of these urinary tests for antibody to H pylori in these patients.METHODS: Serum samples from 61 inpatients with pulmonary tuberculosis were tested using enzyme immunoassay, and urine samples were assayed by enzyme-linked immunosorbent assay method (URINELISA) and immunochromatography method (RAPIRAN). Medicines prescribed to the patients were recorded for medical charts, to evaluate the influences on the results of urinary tests.RESULTS: The sensitivity, specificity, and consistency of URINELISA against the serum test were 93.1%, 65.6%, and 78.6% respectively, and those of RAPIRAN were 86.2%,93.7%, and 90.1% respectively, which were almost equal to the data previously reported. Prescribed medicines had little influence on the results.CONCLUSION: The two urinary tests for detecting H pylori antibody have a diagnostic accuracy in patients with pulmonary tuberculosis given multiple anti-tuberculosis drugs.

  5. The diagnostic sensitivity of dengue rapid test assays is significantly enhanced by using a combined antigen and antibody testing approach.

    Directory of Open Access Journals (Sweden)

    Scott R Fry

    2011-06-01

    Full Text Available BACKGROUND: Serological tests for IgM and IgG are routinely used in clinical laboratories for the rapid diagnosis of dengue and can differentiate between primary and secondary infections. Dengue virus non-structural protein 1 (NS1 has been identified as an early marker for acute dengue, and is typically present between days 1-9 post-onset of illness but following seroconversion it can be difficult to detect in serum. AIMS: To evaluate the performance of a newly developed Panbio® Dengue Early Rapid test for NS1 and determine if it can improve diagnostic sensitivity when used in combination with a commercial IgM/IgG rapid test. METHODOLOGY: The clinical performance of the Dengue Early Rapid was evaluated in a retrospective study in Vietnam with 198 acute laboratory-confirmed positive and 100 negative samples. The performance of the Dengue Early Rapid in combination with the IgM/IgG Rapid test was also evaluated in Malaysia with 263 laboratory-confirmed positive and 30 negative samples. KEY RESULTS: In Vietnam the sensitivity and specificity of the test was 69.2% (95% CI: 62.8% to 75.6% and 96% (95% CI: 92.2% to 99.8 respectively. In Malaysia the performance was similar with 68.9% sensitivity (95% CI: 61.8% to 76.1% and 96.7% specificity (95% CI: 82.8% to 99.9% compared to RT-PCR. Importantly, when the Dengue Early Rapid test was used in combination with the IgM/IgG test the sensitivity increased to 93.0%. When the two tests were compared at each day post-onset of illness there was clear differentiation between the antigen and antibody markers. CONCLUSIONS: This study highlights that using dengue NS1 antigen detection in combination with anti-glycoprotein E IgM and IgG serology can significantly increase the sensitivity of acute dengue diagnosis and extends the possible window of detection to include very early acute samples and enhances the clinical utility of rapid immunochromatographic testing for dengue.

  6. [Evaluation of a Computer-Aided Microscope System and Its Anti-Nuclear Antibody Test Kit for Indirect Immunofluorescence Assay].

    Science.gov (United States)

    Hayashi, Nobuhide; Saegusa, Jun; Uto, Kenichi; Oyabu, Chinami; Saito, Toshiharu; Sato, Itsuko; Kawano, Seiji; Kumagai, Shunichi

    2016-02-01

    Antinuclear antibody (ANA) testing is indispensable for diagnosing and understanding clinical conditions of autoimmune diseases. The indirect immunofluorescence assay (IFA) is the gold standard for ANA screening, and it can detect more than 100 different antibodies, such as anti-PCNA as well as anti-cytoplasmic antibodies. However, complicated procedures of conventional IFA and visual interpretation require highly skilled laboratory staff. This study evaluates the capability, characteristics, and applicability of the recently developed ANA detection system (EUROPattern Cosmic IFA System, EPA) using HEp20-10 cells and the automated pattern recognition microscope. Findings using EPA and conventional methods were compared in 282 sera obtained from connective tissue disease patients and 250 sera from healthy individuals. The concordance of the positivity rate, antibody titer (within +/- 1 tube difference), and the accurate recognition rate of ANA patterns between the automated EPA method and the microscopic judgement of the EPA image by eye was 98.9, 97.4, and 55.3%, respectively. The EPA method showed concordance of the positivity rate as high as 93.3% and concordance of the antibody titer as high as 94.0% (within +/- 1 titer) compared with the conventional method. Regarding the four typical patterns of ANA (homogeneous, speckled, nucleolar, and centromere), large differences between the EPA and conventional methods were not observed, and the rate of concordance between the final EPA result and the conventional method was from 94.1 to 100%. The positivity rate of ANA using the EPA and conventional methods showed marked agreement among the six connective tissue diseases (SLE, MCTD, SSc, PM/DM, and SS) and healthy individuals. Although the EPA system is not considered a complete system and laboratory staff should verify the results, it is a useful system for routine ANA analysis because it contributes to ANA standardization and an efficient workflow. PMID:27311277

  7. Evaluation of tests for rabies antibody and analysis of serum responses after administration of three different types of rabies vaccines.

    Science.gov (United States)

    Grandien, M

    1977-01-01

    Humoral antibody response to three types of rabies vaccines were assayed by the neutralization (NT), the mixed hemadsorption (MH), and the indirect immunofluorescence (IF) tests. The NT and MH tests were used to detect antibodies combining with antigens at the surface of virions and infected cells, whereas the indirect IF test measured antibodies mainly to the rabies nucleocapsid antigen. After immunization with a human diploid cell vaccine, antibodies were detected by both the NT and the MH test in the 14th- and 30th-day serum samples from each of eight vaccinated persons. There was a good correlation between titers obtained with the two tests in this group of vaccinees. Antibodies elicited by duck embryo and nervous tissue vaccines occurred less frequently and in lower titers. In these groups of vaccinees, 5 of 14 and 5 of 10, respectively, had antibodies detectable by the NT test in the 14th- and 30th-day sera but were negative by the MH test. It is suggested that this was due to the high levels of immunoglobulin M antibodies, which are known to be elicited by daily injections of vaccine. Since antibodies of the immunoglobulin M class are considered to be less important for protection against rabies, the MH test is recommended for immunity determinations. Compared with the NT test, this test also offers the advantage of being technically more convenient because of its capacity for testing numerous sera in a single run. Antibody titers obtained by the indirect IF test in the human diploid cell vaccine group were relatively low. Titers in the duck embryo and nervous tissue vaccine groups were higher but did not correlate with the results of the NT test. PMID:323275

  8. A new rapid diagnostic test for detection of anti-Schistosoma mansoni and anti-Schistosoma haematobium antibodies

    Directory of Open Access Journals (Sweden)

    Coulibaly Jean T

    2013-01-01

    Full Text Available Abstract Background Parasitological methods are widely used for the diagnosis of schistosomiasis. However, they are insensitive, particularly in areas of low endemicity, and labour-intensive. Immunoassays based on detection of anti-schistosome antibodies have the merit of high sensitivity and recently a rapid diagnostic test (RDT, incorporating Schistosoma mansoni cercarial transformation fluid (SmCTF for detection of anti-schistosome antibodies in blood has been developed. Here, we assessed the diagnostic performance of the SmCTF-RDT for S. mansoni and S. haematobium infections by comparing it with microscopy for egg detection. Methods A cross-sectional survey was carried out in Azaguié, south Côte d’Ivoire. 118 pre-school-aged children submitted two stool and two urine samples, which were subjected to the Kato-Katz and urine filtration methods for the detection of S. mansoni and S. haematobium eggs, respectively. Urine was also subjected to a commercially available cassette test for S. mansoni, which detects circulating cathodic antigen. A finger-prick blood sample was used for the SmCTF-RDT for detection of anti-S. mansoni and anti-S. haematobium antibodies. Results The prevalence of both anti-S. mansoni and anti-S. haematobium antibodies was more than three times higher than the prevalence of infection estimated by egg detection under a microscope. Using quadruplicate Kato-Katz as the reference standard for the diagnosis of S. mansoni infection, the sensitivity, negative predictive value (NPV, and positive predictive value (PPV of the SmCTF-RDT was 75.0%, 84.2% and 22.5%, respectively. When two urine filtrations were considered as the reference standard for the diagnosis of S. haematobium infection, the sensitivity, NPV and PPV of SmCTF-RDT was 66.7%, 94.9% and 5.1%, respectively. The specificity of SmCTF-RDT, when using egg-detection as the reference standard, was estimated to be 34.4%. This low specificity may be a reflection of the

  9. Guidelines for laboratory testing and result reporting of antibody to hepatitis C virus. Centers for Disease Control and Prevention.

    Science.gov (United States)

    Alter, Miriam J; Kuhnert, Wendi L; Finelli, Lyn

    2003-02-01

    Testing for the presence of antibody to hepatitis C virus (anti-HCV) is recommended for initially identifying persons with hepatitis C virus (HCV) infection (CDC. Recommendations for prevention and control of hepatitis C virus [HCV] infection and HCV-related chronic disease. MMWR 1998;47[No. RR-19] :1-33). Testing for anti-HCV should include use of an antibody screening assay, and for screening test-positive results, a more specific supplemental assay. Verifying the presence of anti-HCV minimizes unnecessary medical visits and psychological harm for persons who test falsely positive by screening assays and ensures that counseling, medical referral, and evaluation are targeted for patients serologically confirmed as having been infected with HCV. However, substantial variation in reflex supplemental testing practices exists among laboratories, and an anti-HCV-positive laboratory report does not uniformly represent a confirmed positive result. These guidelines expand recommendations for anti-HCV testing to include an option for reflex supplemental testing based on screening-test-positive signal-to-cut-off (s/co) ratios. Use of s/co ratios minimizes the amount of supplemental testing that needs to be performed while improving the reliability of reported test results. These guidelines were developed on the basis of available knowledge of CDC staff in consultation with representatives from the Food and Drug Administration and public health, hospital, and independent laboratories. Adoption of these guidelines by all public and private laboratories that perform in vitro diagnostic anti-HCV testing will improve the accuracy and utility of reported anti-HCV test results for counseling and medical evaluation of patients by health-care professionals and for surveillance by public health departments. PMID:12585742

  10. Comparison of saliva and serum for human immunodeficiency virus type 1 antibody testing in Uganda using a rapid recombinant assay.

    OpenAIRE

    Grant, R.M.; Piwowar, E M; Katongole-Mbidde, E; Muzawalu, W; Rugera, S; Abima, J; Stramer, S L; Kataaha, P; Jackson, B.

    1996-01-01

    The accuracy and acceptability of saliva human immunodeficiency virus type 1 (HIV-1) antibody testing were compared with serum testing in a study of paired specimens from HIV-1-seropositive and HIV-1-seronegative Ugandan adults attending a clinic for sexually transmitted diseases. Saliva collection was performed with the Omni-sal device (Saliva Diagnostic Systems, Vancouver, Wash.), and antibody testing was performed by a rapid filter paper assay (Test-Pack; Abbott Laboratories, Abbott Park, ...

  11. Whole-Blood Counting Immunoassay as a Short-Turnaround Test for Detection of Hepatitis B Surface Antigen, Anti-Hepatitis C Virus Antibodies, and Anti-Treponema pallidum Antibodies

    OpenAIRE

    Kudo, Toyoichiro; Kido, Aiko; Nishiyama, Yukiko; Koganeya, Hiroshi; Okuda, Takako; Nabeshima, Motoshige; Iinuma, Yoshitsugu; Ichiyama, Satoshi

    2004-01-01

    Whole-blood samples were used for a counting immunoassay (CIA) with the aim of developing a short- turnaround test. After optimization of the CIA, hepatitis B surface antigen (HBsAg), anti-hepatitis C virus antibodies (anti-HCV), and anti-Treponema pallidum antibodies (anti-TP) were detected as efficiently as by an enzyme immunoassay (EIA) with serum samples. The correlations between whole-blood CIA and serum EIA were 99.8, 97.1, and 99.4% for HBsAg, anti-HCV, and anti-TP, respectively. Whole...

  12. Veterans health administration hepatitis B testing and treatment with anti-CD20 antibody administration

    Science.gov (United States)

    Hunt, Christine M; Beste, Lauren A; Lowy, Elliott; Suzuki, Ayako; Moylan, Cynthia A; Tillmann, Hans L; Ioannou, George N; Lim, Joseph K; Kelley, Michael J; Provenzale, Dawn

    2016-01-01

    AIM: To evaluate pretreatment hepatitis B virus (HBV) testing, vaccination, and antiviral treatment rates in Veterans Affairs patients receiving anti-CD20 Ab for quality improvement. METHODS: We performed a retrospective cohort study using a national repository of Veterans Health Administration (VHA) electronic health record data. We identified all patients receiving anti-CD20 Ab treatment (2002-2014). We ascertained patient demographics, laboratory results, HBV vaccination status (from vaccination records), pharmacy data, and vital status. The high risk period for HBV reactivation is during anti-CD20 Ab treatment and 12 mo follow up. Therefore, we analyzed those who were followed to death or for at least 12 mo after completing anti-CD20 Ab. Pretreatment serologic tests were used to categorize chronic HBV (hepatitis B surface antigen positive or HBsAg+), past HBV (HBsAg-, hepatitis B core antibody positive or HBcAb+), resolved HBV (HBsAg-, HBcAb+, hepatitis B surface antibody positive or HBsAb+), likely prior vaccination (isolated HBsAb+), HBV negative (HBsAg-, HBcAb-), or unknown. Acute hepatitis B was defined by the appearance of HBsAg+ in the high risk period in patients who were pretreatment HBV negative. We assessed HBV antiviral treatment and the incidence of hepatitis, liver failure, and death during the high risk period. Cumulative hepatitis, liver failure, and death after anti-CD20 Ab initiation were compared by HBV disease categories and differences compared using the χ2 test. Mean time to hepatitis peak alanine aminotransferase, liver failure, and death relative to anti-CD20 Ab administration and follow-up were also compared by HBV disease group. RESULTS: Among 19304 VHA patients who received anti-CD20 Ab, 10224 (53%) had pretreatment HBsAg testing during the study period, with 49% and 43% tested for HBsAg and HBcAb, respectively within 6 mo pretreatment in 2014. Of those tested, 2% (167/10224) had chronic HBV, 4% (326/7903) past HBV, 5% (427

  13. Detection of Babesia divergens in southern Norway by using an immunofluorescence antibody test in cow sera

    Directory of Open Access Journals (Sweden)

    Røed Knut H

    2010-10-01

    Full Text Available Abstract Background The incidence of bovine babesiosis, caused by Babesia divergens (Apicomplexa: Piroplasmida has decreased markedly since the 1930 s, but may re-emerge as a consequence of climate change and changes in legislation and pasturing practices. This is a potentially serious disease, with both economical and animal welfare consequences. Therefore, there is a need to survey the distribution of B. divergens. Methods We tested sera from 306 healthy pastured cows from 24 farms along the southern Norwegian coast by using an indirect immunofluorescence IgG antibody test (IFAT. Fractions of seropositive cows were compared by calculating 95% CI. Results The results of this test showed that 27% of the sera were positive for B. divergens antibodies. The fraction of antibody-positive sera that we detected showed a two-humped distribution, with a high fraction of positives being found in municipalities in the western and eastern parts of the study area, while the municipalities between these areas had few or no positive serum samples. Conclusions Neither the farmers' observations nor the Norwegian Dairy Herd Recording System give an adequate picture of the distribution of bovine babesiosis. Serological testing of cows by using IFAT is a convenient way of screening for the presence of B. divergens in an area.

  14. [An evaluation of the China-made HIV antibody test reagents].

    Science.gov (United States)

    Zheng, X W; Zhu, D

    1990-06-01

    This paper reports the results of the evaluation of the China-made HIV antibody screening test reagents, including the IF and IE reagents prepared by the Institute of Virology, CAPM, the ELISA reagent prepared by the Shanghai Institute of Biological Products. Based on the results, the sensitivities of the IF and IE are from 91.2% to 96.9%; the specificities, from 94.6% to 97.3%. Due to the low HIV prevalence in China, the predictive values of negative of these reagents are up to 100%; but the predictive values of positive are very low. It is suggested that these reagents can be used for HIV antibody screen testing in China. The package of some reagents should be improved, the price of some reagents should be decreased. PMID:2390778

  15. Perl Testing A Developer's Notebook

    CERN Document Server

    Langworth, Ian

    2005-01-01

    Is there any sexier topic in software development than software testing? That is, besides game programming, 3D graphics, audio, high-performance clustering, cool websites, et cetera? Okay, so software testing is low on the list. And that's unfortunate, because good software testing can increase your productivity, improve your designs, raise your quality, ease your maintenance burdens, and help to satisfy your customers, coworkers, and managers. Perl has a strong history of automated tests. A very early release of Perl 1.0 included a comprehensive test suite, and it's only improved from th

  16. Serological survey on canine coronavirus antibodies in giant pandas by virus neutralization test

    Institute of Scientific and Technical Information of China (English)

    QIAOJun; XIAXian-zhu; YANGSong-tao; LIDe-sheng; HUGui-xue; GAOYu-wei; SUNHe-ting; ZHAOZhong-pen; XlEZhi-jing; YANFang; HEWen-qi; HUANGGen

    2004-01-01

    In order to survey the infectious situation of canine coronavirus (CCV) in giant panda population, a virus neutralization test detecting specific antibodies against CCV in giant panda's sera was established by using two-fold dilutions of serum and 100 TCID50 of the virus. The 62 sera samples of giant pandas, which were gathered from zoos and reserve region of Sichuan Province, China were detected. The neutralization antibody titer of 1:4 was recognized as the positive criterion, 8 sera samples were detected to be positive, and the positive rate was 12.9%. The titers of neutralizing antibody ranged from 1:8 to 1:32. It was the first comprehensive investigation on neutralization antibodies against CCV in giant panda population in China. The results of study showed that the infection of CCV in giant panda population was universal, which has posed a threat to the health of giant panda. Therefore, it is incumbent on us to study safe and effective vaccines to protect giant panda against CCV infection.

  17. A new Combi test for simultaneous detection of antibodies to viral capsid, early and EBNA antigens of Epstein-Barr virus.

    Science.gov (United States)

    Dobec, M

    1993-06-01

    In order to facilitate the differentiation between a recent (acute) and a past Epstein-Barr virus (EBV) infection, the Combi test was developed. This test is an anticomplement immunofluorescence test (ACIF) requiring only a single serum dilution to be tested on a single cellular spot. The cell line used expresses viral capsid antigen (VCA) and early antigen (EA) in about 5 to 10 percent of the cells as well as EBV nuclear antigens (EBNA) in more than 90 percent of cells. A satisfactory agreement between the Combi test and other tests for antibodies to EBV was obtained (IgG and IgM antibodies to VCA by IFA and EIA and antibodies to EBNA by ACIF including tests for heterophile and complement-fixing antibodies). When the standard serological tests gave negative results, the Combi test was also negative (absence of any fluorescence in the cells). Serologically confirmed recent (acute) infections lead to specific fluorescence in only 5 to 10 percent of the cells, while past infections result in fluorescence in 90 percent or more of the cells. For the diagnosis of a reactivated EBV infection or of EBV-associated malignancies, other tests should be employed. The test is based on the measurement of the activation and specific distribution of the C3 component of complement; the antibody class differentiation is therefore not necessary. The presence of rheumatoid factor (RF) and the IgG competition phenomenon do not influence the results of the Combi test. An introduction of the Combi test will enable a simplified, less expensive and more reliable serodiagnosis of EBV infections. PMID:8394757

  18. Capillary isoelectric focusing method development and validation for investigation of recombinant therapeutic monoclonal antibody.

    Science.gov (United States)

    Suba, Dávid; Urbányi, Zoltán; Salgó, András

    2015-10-10

    Capillary isoelectric focusing (cIEF) is a basic and highly accurate routine analytical tool to prove identity of protein drugs in quality control (QC) and release tests in biopharmaceutical industries. However there are some "out-of-the-box" applications commercially available which provide easy and rapid isoelectric focusing solutions for investigating monoclonal antibody drug proteins. However use of these kits in routine testings requires high costs. A capillary isoelectric focusing method was developed and validated for identification testing of monoclonal antibody drug products with isoelectric point between 7.0 and 9.0. A method was developed providing good pH gradient for internal calibration (R(2)>0.99) and good resolution between all of the isoform peaks (R=2), minimizing the time and complexity of sample preparation (no urea or salt used). The method is highly reproducible and it is suitable for validation and method transfer to any QC laboratories. Another advantage of the method is that it operates with commercially available chemicals which can be purchased from any suppliers. The interaction with capillary walls (avoid precipitation and adsorption as far as possible) was minimized and synthetic isoelectric small molecular markers were used instead of peptide or protein based markers. The developed method was validated according to the recent ICH guideline (Q2(R1)). Relative standard deviation results were below 0.2% for isoelectric points and below 4% according to the normalized migration times. The method is robust to buffer components with different lot numbers and neutral capillaries with different type of inner coatings. The fluoro-carbon coated column was chosen because of costs-effectivity aspects. PMID:26025812

  19. Clinical relevance of multiple antibody specificity testing in anti-phospholipid syndrome and recurrent pregnancy loss.

    Science.gov (United States)

    Tebo, A E; Jaskowski, T D; Hill, H R; Branch, D W

    2008-12-01

    We wanted to evaluate whether testing for anti-phosholipid antibodies other than anti-cardiolipin (aCL) and anti-beta-2 glycoprotein I (abeta2GPI) immunoglobulin (Ig)G and IgM identifies patients with recurrent pregnancy loss (RPL) who may be positive for anti-phospholipid syndrome (APS). In a cross-sectional study comprising 62 patients with APS, 66 women with RPL, 50 healthy blood donors and 24 women with a history of successful pregnancies, we tested IgM and IgG antibodies to phosphatidic acid, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl inositol and phosphatidyl serine with and without beta-2 glycoprotein I (beta2GPI) from a single manufacturer as well as aCL and abeta2GPI antibodies. Diagnostic accuracies of individual and combined anti-phospholipid (aPL) assays were assessed by computing sensitivities, specificities, positive predictive values and negative predictive values together with their 95% confidence intervals. There was a general trend for increased sensitivities in the presence of beta2GPI co-factor with significant effect for certain specificities. The overall combined sensitivity of the non-recommended aPL assays was not significantly higher than that of the aCL and aB2GPI tests. Multiple aPL specificities in RPL group is not significantly different from controls and therefore of no clinical significance. PMID:18826497

  20. Development of a Coxsackievirus A16 neutralization assay based on pseudoviruses for measurement of neutralizing antibody titer in human serum.

    Science.gov (United States)

    Jin, Jun; Ma, Hongxia; Xu, Lin; An, Dong; Sun, Shiyang; Huang, Xueyong; Kong, Wei; Jiang, Chunlai

    2013-02-01

    Serum neutralizing antibody titers are indicative of protective immunity against Coxsackievirus A16 (CV-A16) and Enterovirus 71 (EV71), the two main etiological agents of hand, foot and mouth disease (HFMD), and provide the basis for evaluating vaccine efficacy. The current CV-A16 neutralization assay based on inhibition of cytopathic effects requires manual microscopic examination, which is time-consuming and labor-intensive. In this study, a high-throughput neutralization assay was developed by employing CV-A16 pseudoviruses expressing luciferase for detecting infectivity in rhabdomyosarcoma (RD) cells and measuring serum viral neutralizing antibodies. Without the need to use infectious CV-A16 strains, the neutralizing antibody titer against CV-A16 could be determined within 15h by measuring luciferase signals by this assay. The pseudovirus CV-A16 neutralization assay (pCNA) was validated by comparison with a conventional CV-A16 neutralization assay (cCNA) in testing 174 human serum samples collected from children (age <5 years). The neutralizing antibody titers determined by these two assays were well correlated (R(2)=0.7689). These results suggest that the pCNA can serve as a rapid and objective procedure for the measurement of neutralizing antibodies against CV-A16. PMID:23178532

  1. Preparation of Monoclonal Antibody and Development of Enzyme-linked Immunosorbent Assay Specific for Escherichia coli O157 in Foods

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Objective To prepare monoclonal antibodies (Mab) and antisera specific for Escherichia coli (E.coli) O157, and to develop a sandwich enzyme-linked immunosorbent assay (ELISA) to detect E.coli O157 in foods. Methods Spleen cells from BALB/c mice immunized with the somatic antigen of E.coli O157:H7 were fused with murine Sp2/0 myeloma cells. The hybridoma cell line specific for E.coli O157 was established after having been subcloned. Antisera specific for E.coli O157 was prepared by intravenous injection into New Zealand rabbits with a stain of E.coli O157:H7. The sandwich ELISA was developed with the polyclonal antibody as the capture antibody and the Mab 3A5 as the detection antibody. The inoculated ground poultry meat and pasteurized milk were tested to confirm efficiency of the method. Results Mab 3A5 specific for E.coli O157 and O113:H21 belonged to subtype IgM. The ascetic titers of the antibody was 1:1×106. No cross-reactivity of the Mab was observed with strains of Salmonella spp, Yersinia enterocolitica, Shigella dysenteriae, etc. The purified polyclonal antibody had a titer of 1:1×105 with E.coli O157. The detection limit of this sandwich ELISA was 103-104 cfu E.coli O157/mL in pure culture with a high specificity, which was characterized by every non-O157 strain with negative response. With 10h enrichment procedure, E.coli O157:H7 recovered well from inoculated ground poultry meat and pasteurized milk at levels of 0.1 cfu/g and 0.1 cfu/mL. Conclusion Mab 3A5 specific for E.coli O157 and O113:H21 can be produced by immunizing BALB/c mice with a strain of E.coli O157:H7. Then a sandwich ELISA can be developed with the polyclonal antibody as the capture antibody and the Mab 3A5 as the detection antibody. The method is proved to be a sensitive and specific technique to detect low number of E.coli O157 in food.

  2. Antibodies and Selection of Monoclonal Antibodies.

    Science.gov (United States)

    Hanack, Katja; Messerschmidt, Katrin; Listek, Martin

    2016-01-01

    Monoclonal antibodies are universal binding molecules with a high specificity for their target and are indispensable tools in research, diagnostics and therapy. The biotechnological generation of monoclonal antibodies was enabled by the hybridoma technology published in 1975 by Köhler and Milstein. Today monoclonal antibodies are used in a variety of applications as flow cytometry, magnetic cell sorting, immunoassays or therapeutic approaches. First step of the generation process is the immunization of the organism with appropriate antigen. After a positive immune response the spleen cells are isolated and fused with myeloma cells in order to generate stable, long-living antibody-producing cell lines - hybridoma cells. In the subsequent identification step the culture supernatants of all hybridoma cells are screened weekly for the production of the antibody of interest. Hybridoma cells producing the antibody of interest are cloned by limited dilution till a monoclonal hybridoma is found. This is a very time-consuming and laborious process and therefore different selection strategies were developed since 1975 in order to facilitate the generation of monoclonal antibodies. Apart from common automation of pipetting processes and ELISA testing there are some promising approaches to select the right monoclonal antibody very early in the process to reduce time and effort of the generation. In this chapter different selection strategies for antibody-producing hybridoma cells are presented and analysed regarding to their benefits compared to conventional limited dilution technology. PMID:27236550

  3. EFFICIENCY OF DIFFERENT TECHNIQUES FOR ANTIBODY DETERMINATION AND TESTING OF ANTIGEN-BINDING LYMPHOCYTES IN DIAGNOSTICS OF BRUCELLOSIS

    Directory of Open Access Journals (Sweden)

    B. V. Karalnik

    2014-07-01

    Full Text Available Abstract. The aim of this study was to compare the efficiency of different methods applied for the diagnostics of brucellosis. Huddlson and Wright reactions, RoseBengal test, ELISA-based test system (IgG determination, and a test for antigen-binding lymphocytes (ABL were used. There was shown that the specificity of tests was identical for either reactions based on detection of antibodies with commercial immune reagents and ELISA IgG. The sensitivity of ABL test is, however, superior to all the reactions based on antibody determination. The ABL test has advantages when compared with antibody detection techniques. This method permits early monitoring of brucellosis treatment/its efficiency since early terms, as well as differentiation between positive and false positive results of antibody determination in pregnant womеn.

  4. Development of recombinant antibody technology for application in plant pathogen diagnosis

    OpenAIRE

    Griep, R.A.

    1999-01-01

    This thesis describes the applicability of the novel phage display technique to select plant-pathogen-specific monoclonal antibodies (MAbs) from combinatorial antibody libraries. The retrieved MAbs are so specific that they can be used as diagnostic tools in sensitive immunoassays for the detection and identification of plant pathogens. Testing results, obtained from laboratories that have applied these recombinant MAbs, are discussed in this conclusive chapter.BackgroundIn the last decades, ...

  5. Developing test materials for dyscalculia

    DEFF Research Database (Denmark)

    Lindenskov, Lena; Bent, Lindhardt,

    Aims, requirements and context for the development of test materials for dyscalculia are analyzed. The test materials are to be used for Grade 4 pupils in Danish primary schools. Preliminary results are presented from focus group interview with adolescents and adults, who see themselves as being in...

  6. Developing test materials for dyscalculia

    OpenAIRE

    Lindenskov, Lena; Bent, Lindhardt,

    2015-01-01

    Aims, requirements and context for the development of test materials for dyscalculia are analyzed. The test materials are to be used for Grade 4 pupils in Danish primary schools. Preliminary results are presented from focus group interview with adolescents and adults, who see themselves as being in severe mathematical difficulties.

  7. Validation of serological tests for the detection of antibodies against Treponema pallidum in nonhuman primates.

    Directory of Open Access Journals (Sweden)

    Sascha Knauf

    2015-03-01

    Full Text Available There is evidence to suggest that the yaws bacterium (Treponema pallidum ssp. pertenue may exist in non-human primate populations residing in regions where yaws is endemic in humans. Especially in light of the fact that the World Health Organizaiton (WHO recently launched its second yaws eradication campaign, there is a considerable need for reliable tools to identify treponemal infection in our closest relatives, African monkeys and great apes. It was hypothesized that commercially available serological tests detect simian anti-T. pallidum antibody in serum samples of baboons, with comparable sensitivity and specificity to their results on human sera. Test performances of five different treponemal tests (TTs and two non-treponemal tests (NTTs were evaluated using serum samples of 57 naturally T. pallidum-infected olive baboons (Papio anubis from Lake Manyara National Park in Tanzania. The T. pallidum particle agglutination assay (TP-PA was used as a gold standard for comparison. In addition, the overall infection status of the animals was used to further validate test performances. For most accurate results, only samples that originated from baboons of known infection status, as verified in a previous study by clinical inspection, PCR and immunohistochemistry, were included. All tests, TTs and NTTs, used in this study were able to reliably detect antibodies against T. pallidum in serum samples of infected baboons. The sensitivity of TTs ranged from 97.7-100%, while specificity was between 88.0-100.0%. The two NTTs detected anti-lipoidal antibodies in serum samples of infected baboons with a sensitivity of 83.3% whereas specificity was 100%. For screening purposes, the TT Espline TP provided the highest sensitivity and specificity and at the same time provided the most suitable format for use in the field. The enzyme immune assay Mastblot TP (IgG, however, could be considered as a confirmatory test.

  8. Validation of serological tests for the detection of antibodies against Treponema pallidum in nonhuman primates.

    Science.gov (United States)

    Knauf, Sascha; Dahlmann, Franziska; Batamuzi, Emmanuel K; Frischmann, Sieghard; Liu, Hsi

    2015-03-01

    There is evidence to suggest that the yaws bacterium (Treponema pallidum ssp. pertenue) may exist in non-human primate populations residing in regions where yaws is endemic in humans. Especially in light of the fact that the World Health Organizaiton (WHO) recently launched its second yaws eradication campaign, there is a considerable need for reliable tools to identify treponemal infection in our closest relatives, African monkeys and great apes. It was hypothesized that commercially available serological tests detect simian anti-T. pallidum antibody in serum samples of baboons, with comparable sensitivity and specificity to their results on human sera. Test performances of five different treponemal tests (TTs) and two non-treponemal tests (NTTs) were evaluated using serum samples of 57 naturally T. pallidum-infected olive baboons (Papio anubis) from Lake Manyara National Park in Tanzania. The T. pallidum particle agglutination assay (TP-PA) was used as a gold standard for comparison. In addition, the overall infection status of the animals was used to further validate test performances. For most accurate results, only samples that originated from baboons of known infection status, as verified in a previous study by clinical inspection, PCR and immunohistochemistry, were included. All tests, TTs and NTTs, used in this study were able to reliably detect antibodies against T. pallidum in serum samples of infected baboons. The sensitivity of TTs ranged from 97.7-100%, while specificity was between 88.0-100.0%. The two NTTs detected anti-lipoidal antibodies in serum samples of infected baboons with a sensitivity of 83.3% whereas specificity was 100%. For screening purposes, the TT Espline TP provided the highest sensitivity and specificity and at the same time provided the most suitable format for use in the field. The enzyme immune assay Mastblot TP (IgG), however, could be considered as a confirmatory test. PMID:25803295

  9. An ultra-sensitive monoclonal antibody-based fluorescent microsphere immunochromatographic test strip assay for detecting aflatoxin M1 in milk

    Science.gov (United States)

    A rapid lateral flow fluorescent microspheres immunochromatography test strip (FMs-ICTS) has been developed for the detection of aflatoxin M1 (AFM1) residues in milk. For this purpose, an ultra-sensitive anti-AFM1 monoclonal antibody (MAb) 1D3 was prepared and identified. The IC50 value of the MA...

  10. [Unexpected Diseases in Two Patients with False-Positive Dengue Immunoglobulin M Antibody Test Results].

    Science.gov (United States)

    Matono, Takashi; Kutsuna, Satoshi; Kato, Yasuyuki; Takeshita, Nozomi; Hayakawa, Kayoko; Kanagawa, Shuzo; Ohmagari, Norio

    2016-03-01

    In 2014, an outbreak of 162 domestic dengue fever infections occurred in Tokyo, Japan; the first outbreak of its kind in 70 years. Nineteen of these cases were confirmed in our center. Advancements in diagnostic methods have enabled an earlier diagnosis of dengue fever; however, unfamiliarity with the clinical course and characteristics of diagnostic tests for dengue fever can lead to misdiagnosis. We herein describe 2 cases of Japanese patients with false-positive dengue immunoglobulin M antibody test results, who were finally diagnosed as having dermatomyositis and acute hepatitis A infection, respectively. PMID:27197439

  11. Detection of anti neutrophil antibodies by radio-iodinated protein A binding test

    International Nuclear Information System (INIS)

    The granulocyte associated IgG in normal and neutropenic subjects has been determined by a direct quantitative assay using radiolabeled staphylococcal protein A. This assay allows to postulate an immunological mechanism to explain the neutropenia in 19 cases of neutropenias associated with malfunctions of the immune system and in 4 cases of idiopathic neutropenias. Discussed in this report is the possible interaction of immune complexes bound in vivo to the granulocytes. By an immunofluorescence test, it has been possible to detect IgG or IgM antibodies in only 52% of patients with a positive direct assay. The determination of granulocyte-associated IgG is therefore a better indicator for defining an auto-immune neutropenia than the detection of free antibodies in the sera

  12. An imaging diagnosis of cerebral paragonimiasis: CT and MR findings and correlation with ELISA antibody test

    International Nuclear Information System (INIS)

    To evaluate the CT and MR findings of cerebral paragonimiasis(PW) and to assess the diagnostic value of the specific antibody test by enzyme-linked immunosorbent assay(ELISA) for PW, 55 CT scans and 13 MR images of 57 patients with cerebral PW were reviewed retrospectively, and correlated with the serum/ CSF antibody levels. We divided the patients into three groups, early active (n=21), chronic(n=32), and combined stage(n=4), on the basis of CT/MR findings. In the groups of early active stage the most common and characteristic finding was multiple, conglomerated, ring-like enhancing lesion in the unilateral cerebral hemisphere, which was seen in 52% on CT and 44% on MR. Other non-specific findings included a solitary ring-like or irregular enhancing lesions, ill-defined low density lesions without enhancement, localized hemorrhage with or without enhancing lesions. In the group of chronic stage, there were multiple calcifications of various shapes, most commonly 1-2 cm sized round shape, and associated encephalomalacia. MR was superior to CT in detecting hemorrhage and in characterizing the central contents of ring-shaped calcifications, while it was inferior to CT in identifying small calcifications. Antibody levels of serum and CSF were positive in 86% and 82% in early active group, and in 48% and 31% in chronic stage,retrospectively.The positive rate was significantly different between the two groups (P=0.001). CT/MR findings were characteristic in only approximately half the cases in early active cerebral PW which can be cured by Praziquantel therapy. Therefore, antibody test by ELISA is recommended as a complementary tool, particularly in patients with non-specific imaging findings

  13. An imaging diagnosis of cerebral paragonimiasis: CT and MR findings and correlation with ELISA antibody test

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Kee Hyun; Cha, Sang Hoon; Han, Moon Hee; Kim, Hong Dae; Cho, Seung Yull; Kong, Yoon; Kang, Hyung Keun; Kim, Myung Soon [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1993-05-15

    To evaluate the CT and MR findings of cerebral paragonimiasis(PW) and to assess the diagnostic value of the specific antibody test by enzyme-linked immunosorbent assay(ELISA) for PW, 55 CT scans and 13 MR images of 57 patients with cerebral PW were reviewed retrospectively, and correlated with the serum/ CSF antibody levels. We divided the patients into three groups, early active (n=21), chronic(n=32), and combined stage(n=4), on the basis of CT/MR findings. In the groups of early active stage the most common and characteristic finding was multiple, conglomerated, ring-like enhancing lesion in the unilateral cerebral hemisphere, which was seen in 52% on CT and 44% on MR. Other non-specific findings included a solitary ring-like or irregular enhancing lesions, ill-defined low density lesions without enhancement, localized hemorrhage with or without enhancing lesions. In the group of chronic stage, there were multiple calcifications of various shapes, most commonly 1-2 cm sized round shape, and associated encephalomalacia. MR was superior to CT in detecting hemorrhage and in characterizing the central contents of ring-shaped calcifications, while it was inferior to CT in identifying small calcifications. Antibody levels of serum and CSF were positive in 86% and 82% in early active group, and in 48% and 31% in chronic stage,retrospectively.The positive rate was significantly different between the two groups (P=0.001). CT/MR findings were characteristic in only approximately half the cases in early active cerebral PW which can be cured by Praziquantel therapy. Therefore, antibody test by ELISA is recommended as a complementary tool, particularly in patients with non-specific imaging findings.

  14. Risk to household contacts of tuberculous patientss based on mantoux test and antibody titre

    International Nuclear Information System (INIS)

    Tuberculosis, being an infectious disease, carries a risk of infection to contacts attending tuberculous patients. This study was conducted to evaluate the risk for household contacts of tuberculous patients as compared to non-contacts. The study was conducted at PGMI, Gulab Devi Hospital and Defence Housing Authority Lahore. The study included 120 household contacts and 80 non-contacts. A Cross sectional study for evaluation of antituberculous antibodies levels by ELISA method in two groups; Mantoux positive household contacts 49, Mantoux negative household contacts 71 and normal healthy persons non contacts 80. Routine Haematological investigations like HB, TLC and ESR were done by conventional methods and all the sera of 200 subjects included in the study were tested for IgM, IgG and IgA anti tuberculous antibodies by enzyme linked immunosorbant assay (ELISA). Purified protein derivative 0.1 ml containing 5 TU was injected intradermally. The test was read after 72 hours by measuring the induration around injection site of forearm. There was no difference in the average age of the household contacts and non-contacts. The complaints of pyrexia, night sweats and weight loss were more in house hold contacts as compared to non-contacts. The awareness about BCG vaccination was equal in both. There were 49 contacts with positive Mantoux test while negative Mantoux test was found in 71 contacts. There were only three Mantoux positive among eighty non-contacts. There was no significant difference in the presence of IgM among household contacts as compared to non-contacts. However both IgG and IgA were present in significantly higher number of household contacts compared to non-contacts, household contacts of patients suffering from active pulmonary tuberculosis have more chances of being infected with Mycobacterium tuberculosis as compared to the healthy non-contact, as shown by the higher levels of antituberculous antibodies and positivity of Mantoux test. (author)

  15. Molecular basis of high viscosity in concentrated antibody solutions: Strategies for high concentration drug product development.

    Science.gov (United States)

    Tomar, Dheeraj S; Kumar, Sandeep; Singh, Satish K; Goswami, Sumit; Li, Li

    2016-01-01

    Effective translation of breakthrough discoveries into innovative products in the clinic requires proactive mitigation or elimination of several drug development challenges. These challenges can vary depending upon the type of drug molecule. In the case of therapeutic antibody candidates, a commonly encountered challenge is high viscosity of the concentrated antibody solutions. Concentration-dependent viscosity behaviors of mAbs and other biologic entities may depend on pairwise and higher-order intermolecular interactions, non-native aggregation, and concentration-dependent fluctuations of various antibody regions. This article reviews our current understanding of molecular origins of viscosity behaviors of antibody solutions. We discuss general strategies and guidelines to select low viscosity candidates or optimize lead candidates for lower viscosity at early drug discovery stages. Moreover, strategies for formulation optimization and excipient design are also presented for candidates already in advanced product development stages. Potential future directions for research in this field are also explored. PMID:26736022

  16. Paradoxical role of antibodies in dengue virus infections: considerations for prophylactic vaccine development.

    Science.gov (United States)

    Acosta, Eliana G; Bartenschlager, Ralf

    2016-04-01

    Highly effective prophylactic vaccines for flaviviruses including yellow fever virus, tick-borne encephalitis virus and Japanese encephalitis virus are currently in use. However, the development of a dengue virus (DENV) vaccine has been hampered by the requirement of simultaneous protection against four distinct serotypes and the threat that DENV-specific antibodies might either mediate neutralization or, on the contrary, exacerbate disease through the phenomenon of antibody-dependent enhancement (ADE) of infection. Therefore, understanding the cellular, biochemical and molecular basis of antibody-mediated neutralization and ADE are fundamental for the development of a safe DENV vaccine. Here we summarize current structural and mechanistic knowledge underlying these phenomena. We also review recent results demonstrating that the humoral immune response triggered during natural DENV infection is able to generate neutralizing antibodies binding complex quaternary epitopes only present on the surface of intact virions. PMID:26577689

  17. Development of a recombinant antibody towards PAPP-A for immunohistochemical use in multiple animal species

    DEFF Research Database (Denmark)

    Mikkelsen, Jakob H; Steffensen, Lasse B; Oxvig, Claus

    2014-01-01

    formalin-fixed and paraffin-embedded tissue. For increased sensitivity, affinity maturation to sub-nanomolar affinity was then carried out. The resulting recombinant antibody, PAC1-D8-mIgG2a, detects PAPP-A specifically and sensitively in human tissue. In addition, this antibody allows detection of PAPP...... normal physiology and under different pathological conditions. However, antibodies for the detection of PAPP-A in non-human tissues have been lacking, although needed for use with several animal models which are currently being developed. To develop a monoclonal antibody suitable for the......-A in non-human species. We demonstrate its usefulness for the visualization of PAPP-A in murine and porcine tissues....

  18. Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates

    DEFF Research Database (Denmark)

    Robardet, E.; Andrieu, S.; Rasmussen, Thomas Bruun;

    2013-01-01

    Twelve National Reference Laboratories (NRLs) for rabies have undertaken a comparative assay to assess the comparison of fluorescent antibody test (FAT) results using five coded commercial anti-rabies conjugates (Biorad, Bioveta, Fujirebio, Millipore, and SIFIN conjugates). Homogenized positive...

  19. Application of a human osteogenic sarcoma cell culture for detection of human cytomegalovirus antibody by immunofluorescence tests.

    OpenAIRE

    Furukawa, T; Herold, R; Zolnick, P; Plotkin, S A

    1981-01-01

    A human osteogenic sarcoma cell culture is useful in the immunofluorescence serological test for detecting human cytomegalovirus antibody. These sarcoma cells are chronically infected with human cytomegalovirus and provide a constant number of immunofluorescence-positive cells.

  20. Development of two potential diagnostic monoclonal antibodies against human cytomegalovirus glycoprotein B

    Directory of Open Access Journals (Sweden)

    Fan Jun

    2012-07-01

    Full Text Available Human cytomegalovirus glycoprotein B (gB represents a target for diagnosis and treatment in view of the role it plays in virus entry and spread. Nevertheless, to our knowledge, rare detection of a gB antigen has been reported in transplant patients and limited information is available about diagnostic gB monoclonal antibodies (mAbs. Our aim was to develop gB mAbs with diagnostic potential. Hydrophilic gB peptides (ST: amino acids 27-40, SH: amino acids 81-94 of favorable immunogenicity were synthesized and used to immunize BALB/c mice. Two mAbs, named ZJU-FH6 and ZJU-FE6, were generated by the hybridoma technique and limited serial dilution and then characterized by indirect ELISA, Western blotting, immunoprecipitation, and immunohistochemical staining. The mAbs displayed high titers of specific binding affinities for the ST and SH synthetic peptides at an mAb dilution of 1:60,000 and 1:240,000, respectively. Western blotting and immunoprecipitation indicated that these mAbs recognized both denatured and native gB of the Towne and AD169 strains. The mAbs, when used as the primary antibody, showed positive staining in cells infected with both Towne and AD169 strains. The mAbs were then tested on patients submitted to allogeneic hematopoietic stem cell transplantation. The gB antigen positivity rates of the patients tested using ZJU-FH6 and ZJU-FE6 were 62.0 and 63.0%, respectively. The gB antigen showed a significant correlation with the level of pp65 antigen in peripheral blood leukocytes. In conclusion, two potential diagnostic gB mAbs were developed and were shown to be capable of recognizing gB in peripheral blood leukocytes in a reliable manner.

  1. Development of an antigen microarray for high throughput monoclonal antibody selection

    OpenAIRE

    Staudt, Nicole; Müller-Sienerth, Nicole; Wright, Gavin J.

    2014-01-01

    Monoclonal antibodies are valuable laboratory reagents and are increasingly being exploited as therapeutics to treat a range of diseases. Selecting new monoclonal antibodies that are validated to work in particular applications, despite the availability of several different techniques, can be resource intensive with uncertain outcomes. To address this, we have developed an approach that enables early screening of hybridoma supernatants generated from an animal immunised with up to five differ...

  2. Cask development, testing, and licensing

    International Nuclear Information System (INIS)

    The NuPac 125-B Rail Cask was developed to provide a safe means of transporting the damaged core of Three Mile Island Unit 2 from the TMI site at Middletown, PA, to the Idaho National Engineering laboratory (INEL) at Idaho Falls, ID. The development of the NuPac 125-B Rail Cask posed two engineering and technical management challenges; Licensing Strategy - The NuPac 125-B Rail Cask represented the first irradiated fuel rail cask developed within the United States in the past decade, a decade characterized by changing nuclear regulations, and Accelerated Schedule - The TMI-2 defueling schedule demanded a cask development schedule one-third as long as normally required. These challenges governed the overall development and licensing process for the cask. First, a high degree of conservation was incorporated into the design to allow quick, simplified demonstrations of adequacy to regulatory staff. Second, redundant design techniques were employed in all areas of uncertainty. The testing program eliminated performance uncertainties and validated predictions and predictive models. Drop tests of a quarter-scale model of the cask were conducted, and results were correlated with analytic predictions to verify structural and mechanical performance of the cask. Full-scale tests of the canisters were conducted to verify structural behavior of canister internals which provide criticality control. This paper describes the testing program for the NuPac 125-B Rail Cask, presents results therefrom, and correlates findings with Regulation 10 CFR 71 of the U.S. Nuclear Regulatory Commission

  3. Development of monoclonal antibodies against parathyroid hormone: Genetic control of the immune response to human PTH

    International Nuclear Information System (INIS)

    The authors embarked upon a program to develop monoclonal antibodies to the biologically active amino terminal region of PTH. Using the BALB/c mouse for immunization, fully biologically active synthetic human PTH-(1-34) and bovine PTH-(1-84) as immunogens, monoclonal antibody methods and a solid-phase screening assay in which PTH-(1-34) was adhered to polyvinylchloride plates in a manner that preserved immunoreactivity. They generated 17 monoclonal antibodies against the amino-terminal portion of parathyroid hormone. Isotypic analysis of these monoclonal antibodies was performed using affinity purified goat anti-mouse immunoglobins specific for IgG heavy chains, γ/sub 1/, γ/sub 2a/, γ/sub 2b/, γ/sub 3/; α(IgA); and μ(Igm). All antibodies were IgM as evidenced by 40 times greater than background radioactivity when 25,000 cpm of /sup 125/I-labeled goat anti-mouse IgM was used as second antibody in a solid-phase radioimmunoassay. All incubations with iodinated second antibodies to other heavy chain classes of immunoglobins demonstrated background radioactivity. Extensive synthetic work in the laboratory for multiple biologic studies of structure-activity relationships of PTH, as well as analog design, has led to the synthesis of many peptide analogues and fragments from 7 to 34 amino acids in length. Study of the antibody recognition site (region specificity) by two of these monoclonal antibodies, 10A/sub 7/, and 6B/sub 1/, was undertaken with synthetic peptides

  4. Biosimilar monoclonal antibodies: preclinical and clinical development aspects.

    Science.gov (United States)

    Gonçalves, João; Araújo, Filipe; Cutolo, Maurizio; Fonseca, João Eurico

    2016-01-01

    Biological drugs and their originated biosimilars are large, highly complex molecules derived from living cells or organisms. Traditional medicines, by contrast, are usually simple molecules of low molecular weight, synthesised by chemical means. The distinct complexities and methods of manufacture create an important difference between biosimilars and conventional generic drugs: while chemical generics can be fully characterised as identical to the originator product, biosimilars cannot. In addition, biological therapies are inherently variable, creating unavoidable differences between even subsequent batches of the same product. An expiring patent does not necessarily mean that the manufacturing process of the originator product becomes available to the biosimilar developers (for instance, the relevant cell line clone and growth medium). Therefore, it cannot be guaranteed that biosimilar products are identical to their reference product on a molecular level. This difference has important implications for the regulation and licensing of biosimilars. While conventional generic drugs require only a limited comparison and demonstration of identical chemical structure to the reference product, biosimilars require far more rigorous testing. In general, there must be a thorough comparison of structural and functional characteristics between biosimilar and originator drug. Stepwise nonclinical in vitro and in vivo approaches are recommended to evaluate the similarity of both drugs and any identified micro-heterogeneities must then be assessed for their impact on safety and clinical performance. Subsequently, clinical pharmacokinetic (PK) studies need to be performed in order to demonstrate a similar PK profile, prior to conducting clinical efficacy trials. PMID:27383278

  5. Evaluation of a direct immunofluorescent antibody (difma test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis

    Directory of Open Access Journals (Sweden)

    Martha E. Chico

    1995-06-01

    Full Text Available A direct immunofluorescent antibody (DIFMA test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

  6. Interlaboratory testing of porcine sera for antibodies to porcine circovirus type 2

    DEFF Research Database (Denmark)

    McNair, I.; Marshall, M.; McNeilly, F.; Bøtner, Anette; Ladekjær-Mikkelsen, A.S.; Vincent, I.; Herrmann, Brigitte; Sanchez, R.; Rhodes, C.

    2004-01-01

    A panel of 20 porcine sera was distributed to 5 laboratories across Europe and Canada. Each center was requested to test the sera for the presence of porcine circovirus type 2 antibodies using the routine assays, indirect immunofluorescence assay (IFA) and indirect immunoperoxidase monolayer assay...... (IPMA), and to determine the titer of each serum. Results from all centers were then compiled and correlated. They demonstrate a wide variation in the titers obtained between laboratories. These differences were dependent on the assay used and the choice of fixative. In general, IPMA gave higher titers...

  7. Evaluation of five commercial tests for detection of immunoglobulin M antibodies to human parvovirus B19.

    OpenAIRE

    Bruu, A L; Nordbø, S A

    1995-01-01

    The following commercial tests for detection of immunoglobulin M antibodies to human parvovirus B19 were evaluated: Ideia Parvovirus B19-IgM, MRL Diagnostics Human Parvovirus B19 IgM ELISA, Parvoscan-B19, and Biotrin Parvo B19 IgM EIA and IF. A total of 203 serum specimens from patients who probably have current B19 infections or have other viral infections and sera with rheumatoid factor were investigated. Between 75 and 79 of 102 serum samples from patients thought to have current B19 infec...

  8. Enhanced performance of an innovative dengue IgG/IgM rapid diagnostic test using an anti-dengue EDI monoclonal antibody and dengue virus antigen

    OpenAIRE

    Jihoo Lee; Young-Eun Kim; Hak-Yong Kim; Mangalam Sinniah; Chom-Kyu Chong; Hyun-Ok Song

    2015-01-01

    High levels of anti-dengue IgM or IgG can be detected using numerous rapid diagnostic tests (RDTs). However, the sensitivity and specificity of these tests are reduced by changes in envelope glycoprotein antigenicity that inevitably occur in limited expression systems. A novel RDT was designed to enhance diagnostic sensitivity. Dengue viruses cultured in animal cells were used as antigens to retain the native viral coat protein. Monoclonal antibodies (mAbs) were then developed, for the first ...

  9. An Enhanced Pre- and Postnatal Development Study in Cynomolgus Monkeys with Tabalumab: A Human IgG4 Monoclonal Antibody.

    Science.gov (United States)

    Breslin, William J; Hilbish, Kim G; Martin, Jennifer A; Halstead, Carolyn A; Newcomb, Deanna L; Chellman, Gary J

    2015-06-01

    Tabalumab, a human IgG4 monoclonal antibody (mAb) with neutralizing activity against both soluble and membrane B-cell activating factor (BAFF), has been under development for the treatment of autoimmune diseases. The purpose of this study was to determine the potential adverse effects of maternal tabalumab exposure on pregnancy, parturition, and lactation of the mothers and on the growth, viability, and development of the offspring through postnatal day (PND) 204. Tabalumab was administered by subcutaneous injection to presumed pregnant cynomolgus monkeys (16-19 per group) every 2 weeks from gestation day (GD) 20 to 22 until parturition at doses of 0, 0.3, or 30 mg/kg. Evaluations in mothers and infants included clinical signs, body weight, toxicokinetics, blood lymphocyte phenotyping, T-cell-dependent antibody response (infants only), antitherapeutic antibody (ATA), organ weights (infants only), and gross and microscopic histopathology. Infants were also examined for external and visceral morphologic and neurobehavioral development. There were no adverse tabalumab-related effects on maternal or infant endpoints. An expected pharmacological decrease in peripheral blood B-lymphocytes occurred in adults and infants; however, B-cell recovery was evident by PND154 in adults and infants at 0.3 mg/kg and by PND204 in infants at 30 mg/kg. At 30 mg/kg, a reduced IgM antibody response to T-cell-dependent antigen keyhole limpet hemocyanin (KLH) was observed following primary immunization. Following secondary KLH immunization, all infants in both dose groups mounted anti-KLH IgM and IgG antibody responses similar to control. Placental and mammary transfer of tabalumab was demonstrated. In conclusion, the no-observed-adverse-effect level for maternal and developmental toxicity was 30 mg/kg, the highest dose tested. Exposures at 30 mg/kg provide a margin of safety of 16× the anticipated clinical exposure. PMID:26195230

  10. Development of a Recombinant Antibody with Specificity for Chelated Uranyl Ions

    International Nuclear Information System (INIS)

    The goal of our project is to continue the development of new techniques for rapid, automated identification of radionuclides, metals, and chelators that may contaminant sur face and groundwater at DOE sites. One of the four specific aims of the present project is to develop new technologies in antibody engineering that will enhance our immunosensor program. Recombinant antibodies have potential advantages over monoclonal antibodies produced by standard hybridoma technology. The cloned genes represent a stable, recoverable source for antibody production. In addition, the recombinant format offers opportunities for protein engineering that enhances antibody performance and for studies that relate antibody sequence to binding activity. In this study, a hybridoma that synthesized an antibody (12F6) that recognized a 1:1 complex between 2,9-dicarboxyl-1,10- phenanthroline (DCP) and UO22+ was used as a source of RNA for the development of a recombinant (Fab)2 fragment. RNA was isolated from the 12F6 hybridoma and the cDNA encoding the entire κ light chain and the linked VH and C1 portions of the heavy chain were amplified from total RNA. cDNA sequences were verified by comparison with the N-terminal amino acid sequences of the light and heavy chains of the native 12F6 monoclonal antibody. A leader sequence and appropriate restriction sites were added to each chain, and the fragments were ligated into a commercial dicistronic vector (pBudCE4.1, Invitrogen, Inc.). COS-1 cells were transfected with this vector and the culture supernatant was assayed for activity and the (Fab)2 protein. Cells transfected with vector containing 12F6 cDNA synthesized and secreted recombinant (Fab)2 fragments that bound to the UO22+-DCP complex with an affinity indistinguishable from that of a (Fab)2 fragment prepared from the native antibody. Molecular models of the heavy and light chain variable domains were constructed according to the canonical structures method detailed by Morea et al

  11. Development of a Recombinant Antibody with Specificity for Chelated Uranyl Ions

    Energy Technology Data Exchange (ETDEWEB)

    X. Li; A.M. Kriegel; T.C. Bishop; R.C. Blake; E. Figueiredo; H. Yu; D.A. Blake

    2005-04-18

    The goal of our project is to continue the development of new techniques for rapid, automated identification of radionuclides, metals, and chelators that may contaminant sur face and groundwater at DOE sites. One of the four specific aims of the present project is to develop new technologies in antibody engineering that will enhance our immunosensor program. Recombinant antibodies have potential advantages over monoclonal antibodies produced by standard hybridoma technology. The cloned genes represent a stable, recoverable source for antibody production. In addition, the recombinant format offers opportunities for protein engineering that enhances antibody performance and for studies that relate antibody sequence to binding activity. In this study, a hybridoma that synthesized an antibody (12F6) that recognized a 1:1 complex between 2,9-dicarboxyl-1,10- phenanthroline (DCP) and UO{sub 2}{sup 2+} was used as a source of RNA for the development of a recombinant (Fab){sub 2} fragment. RNA was isolated from the 12F6 hybridoma and the cDNA encoding the entire {kappa} light chain and the linked VH and C1 portions of the heavy chain were amplified from total RNA. cDNA sequences were verified by comparison with the N-terminal amino acid sequences of the light and heavy chains of the native 12F6 monoclonal antibody. A leader sequence and appropriate restriction sites were added to each chain, and the fragments were ligated into a commercial dicistronic vector (pBudCE4.1, Invitrogen, Inc.). COS-1 cells were transfected with this vector and the culture supernatant was assayed for activity and the (Fab){sub 2} protein. Cells transfected with vector containing 12F6 cDNA synthesized and secreted recombinant (Fab){sub 2} fragments that bound to the UO{sub 2}{sup 2+}-DCP complex with an affinity indistinguishable from that of a (Fab){sub 2} fragment prepared from the native antibody. Molecular models of the heavy and light chain variable domains were constructed according to the

  12. Development and characterization of a monoclonal antibody to human embryonal carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Khazaeli, M.B.; Beierwaltes, W.H.; Pitt, G.S.; Kabza, G.A.; Rogers, K.J.; LoBuglio, A.F.

    1987-06-01

    A monoclonal anti-testicular carcinoma antibody was obtained via the somatic cell fusion technique by immunization of BALB/c mice with freshly prepared single cell suspension from a patient with testicular embryonal carcinoma with choriocarcinoma components. The hybridoma supernates were screened against the testicular carcinoma cells used in the immunization as well as normal mononuclear white blood cells isolated from the same patient. An antibody (5F9) was selected which bound to fresh tumor cells from two patients with embryonal testicular carcinoma and failed to bind to fresh tumor cells from 24 patients (2 seminoma, 2 melanoma, 3 neck, 2 esophageal, 1 ovarian, 3 colon, 1 prostate, 2 breast, 1 liposarcoma, 3 endometrial, 1 kidney, 1 adrenal, 1 larynx and 1 bladder tumors) or cell suspensions prepared from normal liver, lung, spleen, ovary, testes, kidney, red blood cells or white blood cells. The antibody was tested for its binding to several well established cancer cell lines, and was found to bind to the BeWo human choriocarcinoma and two human embryonal carcinoma cell lines. The antibody did not react with 22 other cell lines or with hCG. The antibody was labeled with /sup 131/I and injected into nude mice bearing BeWo tumors and evaluated for tumor localization by performing whole body scans with a gamma camera 5 days later. Six mice injected with the antibody showed positive tumor localization without the need for background subtraction while six mice injected with MOPC-21, a murine myeloma immunoglobulin, demonstrated much less tumor localization. Tissue distribution studies performed after scanning showed specific tumor localization (8:1 tumor: muscle) for the monoclonal antibody and no specific localization for MOPC-21.

  13. Serum auto-antibody testing for early diagnosis of breast cancer

    International Nuclear Information System (INIS)

    The aim of this thesis is generate prototype-tests suitable for randomized prospective validation of auto-antibody based diagnostic testing using serum samples. Tumours can stimulate the production of auto-antibodies against autologous cellular proteins known as TAAs (tumour associated antigens). This discovery has lead to a possibility of using the auto-antibodies as serological tools for the early diagnosis and management of breast cancer. The recombinant proteins expressed by the SEREX clones, identified from screenings of brain and lung tumour, were used for the production of the protein microarrays and macroarrays. The protein microarrays showed better correlation between the replicates of the serum samples used. The optimized protocols were used for the subsequent experiments. A sizable panel of 642 clone-proteins was selected by marker-screening on protein macroarrays with 38000 clones. These 642 clone-proteins were used to generate protein microarrays that differentiated serum samples from breast cancer patients and controls. Antigenic peptide motifs were identified by in-silico analysis of 642 clone-proteins and peptide arrays were generated using synthetically generated peptides. Comparative studies between protein microarrays and peptide microarrays were done using breast cancer and healthy control samples. Simultaneously, SEREX strategy was used for the identification of the immunogenic TAAs. I identified 192 cDNA expression clones derived from breast cancer tissue samples and the selection was done using breast cancer sera. The genes corresponding to these clones were found over-represented for the pathways that are known to be associated with cancers. These genes showed typical features of TAAs, like over-expression, mutations and fusion genes. (author)

  14. Choosing wisely: Review and commentary on anti-nuclear antibody (ANA) testing.

    Science.gov (United States)

    Fritzler, Marvin J

    2016-03-01

    Choosing Wisely®: Next Steps in Improving Healthcare Value is an initiative of the American Board of Internal Medicine (ABIM) Foundation. The driving forces for the Choosing Wisely (CW) campaign include rising and unstainable health care expenditures and evidence that there is lack of fiscal stewardship of health care resources. The American College of Rheumatology and the Canadian Rheumatology Association published their top five Choosing Wisely recommendations, the first of which pertained to antinuclear antibodies (ANA) and ANA subserology testing. Concerns about the wasteful use of these tests prompted an analysis of the expenditures attributable to ANA testing as a proportion of total health care expenditures and based on a financial model was in the range of 0.00125%. It is suggested that if the sole use of ANA testing is to add evidence to support a diagnosis when the pre-test probability is high, then the ANA test has limited clinical value. Accordingly, the goal of ANA testing needs to be reconsidered and expanded beyond an approach to simply confirming a diagnosis with 'intention to treat' to a goal of case finding of 'pre- or early disease' with an 'intent to prevent' disease. This an area where more significant inroads can be made in preventing end organ disease and thereby reducing health care expenditures HCE. One CW recommendation that bears emphasizing is that, with a few possible exceptions, repeat ANA or ANA subserology testing has little clinical value in monitoring disease activity or predicting a flare. PMID:26687321

  15. Development of QCM Biosensor with Specific Cow Milk Protein Antibody for Candidate Milk Adulteration Detection

    Directory of Open Access Journals (Sweden)

    Setyawan P. Sakti

    2016-01-01

    Full Text Available Adulteration of goat milk is usually done using cow’s milk product. Cow milk is used as it is widely available and its price is cheaper compared to goat milk. This paper shows a development of candidate tools for milk adulteration using cow milk. A quartz crystal microbalance immunosensor was developed using commercial crystal resonator and polyclonal antibody specific to cow milk protein. A specific protein at 208 KDa is found only in cow milk and does not exist in goat milk. The existence of this protein can be used as an indicator of cow milk content in a target solution. To detect the PSS 208 kDa protein, antibody specific to the PSS 208 was developed. The purified antibody was immobilized on top of the sensor surface on a polystyrene layer. The fraction of the immobilized antibody on the sensor was found at 1.5% of the given antibody. Using a static reaction cell, the developed immunosensor could detect the specific cow milk protein in buffer solution. The detection limit is 1 ppm. A linear relationship between frequency change and specific protein of cow milk concentration is found from a concentration of 1 ppm to 120 ppm.

  16. Selection of matched pair of monoclonal antibodies for development of immunoradiometric assay (IRMA) : our experience with IRMA of TSH

    International Nuclear Information System (INIS)

    Full text: In immunoradiometricassay (IRMA) two antibodies raised against two different epitopes of the same antigen are used, one bound to a solid phase (capture antibody) and the other labelled with 125I (detector antibody). The development of any IRMA thus involves proper selection of the capture and detector antibody, preparation of solid phase, labelling of the antibody and assay optimization. Extensive studies have been carried out on these aspects in our laboratory with greater emphasis on the behavior of different pairs of antibodies as sandwich partners : monoclonal-monoclonal and monoclonal-polyclonal antibodies. The parameters studied include the ease of radio-iodination of different monoclonal antibodies, the effect of interchange of capture and detector antibody etc. Keeping TSH antibody as a model, two different monoclonal antibodies, a polyclonal antibody and a tracer from a commercial TSH IRMA kit were used in this study. Based on our studies an assay procedure for in-house IRMA of TSH has been developed with a sensitivity of 0.1 μIU/ml and validated

  17. Enhancement of Antibody Titre and Development of Additional Red Cell Alloantibodies Following Intrauterine Transfusion.

    Science.gov (United States)

    Dubey, Anju; Sonker, Atul; Chaudhary, Rajendra

    2016-03-01

    Intrauterine blood transfusion is the mainstay of managing foetuses with severe anemia. It may however result in fetomaternal hemorrhage, which in cases of Rh isoimmunisation may increase the severity of the disease by enhancing the maternal immunological response to fetal antigens. This study was conducted to determine the frequency, specificity and origin of additional red cell antibodies which developed after IUT. The change in the titre of allo anti-D following IUT was also determined. Antibody detection and titration was done on the blood samples of all the patients before and after intrauterine blood transfusion to check for the development of additional antibody and change in the titre of existing anti-D. Severe anemia was found in 17 (58.6 %) fetuses who received a total of 42 ultrasound-guided IUTs. Development of antibodies additional to anti-D in maternal serum was seen in 5 (29.4 %) cases. The specificity of additional alloantibodies was anti-C in four cases whereas it was anti-E in one case. Four fold or greater increase in existing allo-anti D titre was seen in 6 (35.3 %) cases after IUT. Enhancement of maternal sensitisation leading to an increase in maternal antibody titre is particularly seen after the first IUT. Matching of the donor RBCs particularly for Rh antigens might prevent the induction of additional alloantibodies against these antigens. IUT as a treatment modality should be given judiciously and only when the need is inevitable. PMID:26855513

  18. Development and immunochemical evaluation of a novel chicken IgY antibody specific for KLK6

    Directory of Open Access Journals (Sweden)

    Sotiropoulou Georgia

    2012-12-01

    Full Text Available Abstract Background Human kallikrein-related peptidase 6 (KLK6 has been implicated in various types of cancer and in neurodegenerative and demyelinating diseases including multiple sclerosis. Further, anti-KLK6 antibodies attenuated disease manifestations in the mouse model of multiple sclerosis. Availability of specific antibodies against KLK6 is fundamental to the development of improved diagnostic and/or immunotherapeutic applications. Here, we exploited the enhanced immunogenicity of mammalian proteins in avian species to generate a polyclonal antibody against KLK6. Results Chicken were immunized with recombinant KLK6 and antibodies Y (IgYs were purified from egg yolk with a simple procedure and evaluated for KLK6 detection by ELISA and Western blot using recombinant proteins and human cell lysates and supernatants. The anti-KLK6 Y polyclonal exhibited high affinity for KLK6 with a detection limit of 30 fmol. On the other hand, the widely used rabbit polyclonal antibody that was raised against the same recombinant KLK6 had a detection limit of 300 fmol. Moreover, the IgYs did not display any crossreactivity with recombinant KLKs or endogenous KLKs and other cellular proteins. Conclusions Based on its high specificity and sensitivity the developed anti-KLK6 IgY is expected to aid the development of improved diagnostic tools for the detection of KLK6 in biological and clinical samples.

  19. Antibodies to Lactobacilli and Bifidobacteria in Young Children with Different Propensity to Develop Islet Autoimmunity

    Directory of Open Access Journals (Sweden)

    Ija Talja

    2014-01-01

    Full Text Available The intestinal microbiota is essential to the maturation and homeostasis of the immune system. Immunoblot assays were used to establish the prevalence of serum IgG, IgM, and IgA antibodies specific for Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus GG proteins in young children presenting with or without type 1 diabetes (T1D. We demonstrated that children between the ages of 6 and 12 months had a substantial increase in the frequency of IgG antibodies specific for L. rhamnosus GG proteins. We measured IgG, IgM, and IgA class antibody reactivity against B. adolescentis DSM 20083, B. adolescentis DSM 20086, and B. longum DSM 20088 proteins demonstrating significantly higher IgA responses against B. adolescentis DSM 20083 strain proteins in children who developed islet autoimmunity and T1D later in life. B. adolescentis strains showed more IgM type antibodies in children who developed T1D later in life, but the difference was not statistically significant. B. longum proteins were recognized by IgG and IgA antibodies to a higher extent compared to other bacteria studied. These results confirm that differences in immune reactivity against some commensal strains in young children may represent a different risk factor for developing T1D.

  20. Antibodies to Lactobacilli and Bifidobacteria in young children with different propensity to develop islet autoimmunity.

    Science.gov (United States)

    Talja, Ija; Kubo, Anna-Liisa; Veijola, Riitta; Knip, Mikael; Simell, Olli; Ilonen, Jorma; Vähä-Mäkilä, Mari; Sepp, Epp; Mikelsaar, Marika; Utt, Meeme; Uibo, Raivo

    2014-01-01

    The intestinal microbiota is essential to the maturation and homeostasis of the immune system. Immunoblot assays were used to establish the prevalence of serum IgG, IgM, and IgA antibodies specific for Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus GG proteins in young children presenting with or without type 1 diabetes (T1D). We demonstrated that children between the ages of 6 and 12 months had a substantial increase in the frequency of IgG antibodies specific for L. rhamnosus GG proteins. We measured IgG, IgM, and IgA class antibody reactivity against B. adolescentis DSM 20083, B. adolescentis DSM 20086, and B. longum DSM 20088 proteins demonstrating significantly higher IgA responses against B. adolescentis DSM 20083 strain proteins in children who developed islet autoimmunity and T1D later in life. B. adolescentis strains showed more IgM type antibodies in children who developed T1D later in life, but the difference was not statistically significant. B. longum proteins were recognized by IgG and IgA antibodies to a higher extent compared to other bacteria studied. These results confirm that differences in immune reactivity against some commensal strains in young children may represent a different risk factor for developing T1D. PMID:24741589

  1. High-Throughput Testing of Antibody-Dependent Binding Inhibition of Placental Malaria Parasites

    DEFF Research Database (Denmark)

    Nielsen, Morten A; Salanti, Ali

    2015-01-01

    different human receptors through Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) on the surface of the infected cell. As the var genes encoding the large PfEMP1 antigens are extensively polymorphic, vaccine development strategies are focused on targeting the functional binding epitopes. This...... involves identification of recombinant fragments of PfEMP1s that induce antibodies, which hinder the adhesion of the IE to a given receptor or tissue. Different assays to measure the blocking of adhesion have been described in the literature, each with different advantages. This chapter describes a high...

  2. Correlation of serum antithyroid microsomal antibody and autologous serum skin test in patients with chronic idiopathic urticaria

    OpenAIRE

    Snehal Balvant Lunge; Milind Borkar; Sushil Pande

    2015-01-01

    Background: About 25–45% of patients of chronic urticaria (CU) have been stated to have histamine releasing autoantibodies in their blood. The term autoimmune urticaria is increasingly being accepted for this subgroup of patients. Review of the literature suggests high autologous serum skin test (ASST) positivity and presence of antithyroid microsomal antibodies in patients with autoimmune urticaria. Aims: To study prevalence of ASST positivity and antithyroid microsomal antibodies in chronic...

  3. Plaque reduction neutralization antibody test does not accurately predict protection against dengue infection in Ratchaburi cohort, Thailand

    OpenAIRE

    Sirivichayakul, Chukiat; Sabchareon, Arunee; Limkittikul, Kriengsak; Yoksan, Sutee

    2014-01-01

    Background The plaque reduction neutralization test (PRNT) is currently the best and most widely accepted approach to measuring virus-neutralizing and protective antibodies to dengue virus, and in assessing the immunogenicity of a dengue vaccine. However, the correlation between presence of dengue-neutralizing antibody and protection from infection is not absolute. Findings In a cohort study in Ratchaburi Province, Thailand, 48 subjects with serologically confirmed symptomatic dengue infectio...

  4. A large-scale radiometric micro-quantitative complement fixation test for serum antibody titration

    International Nuclear Information System (INIS)

    A micro-quantitative complement fixation (CF) procedure based on 51Cr release is described. The method employs 50% hemolysis as end point and the alternation equation to calculate the amount of complement involved in the hemolytic reaction. Compared to the conventional CF tests, the radiometric procedure described here is very precise and consistently reproducible. Also, since only 3 4-fold dilutions of sera are used for the titration of antibodies over a wide range of concentrations, the test is very concise and is economical to perform. Its format is amenable to automation and computerization. This radioimetric CF procedure is thus most useful for large-scale immunological research and epidemiological surveilance studies. (Auth.)

  5. Demystifying the Positive Antinuclear Antibody Test in Children: A Clinical Review.

    Science.gov (United States)

    Rodriguez, Martha; Tesher, Melissa S; Wagner-Weiner, Linda

    2015-06-01

    A 15-year-old girl presented with knee pain, associated with a positive antinuclear antibody (ANA). She denied joint swelling or morning stiffness and remained physically active despite the pain. A physical examination was unremarkable except for articular hypermobility. Laboratory results were also unremarkable. Therefore, the positive ANA was determined to be nonspecific, and not concerning. In the evaluation of children with musculoskeletal complaints, unusual rash, or fatigue, an ANA assessment is frequently considered. When is this test most likely to be useful? What is the appropriate follow up for a positive result? Which results are concerning for an autoimmune process? This article reviews the literature to address these practical concerns. Understanding the indications for ordering an ANA, and the correct interpretation of a positive ANA, may reduce unnecessary referrals and costly tests. Moreover, the misperception that a positive ANA indicates a rheumatologic disease can cause significant patient and parental anxiety. PMID:26114367

  6. Isolation of additional monoclonal antibodies directed against cell surface antigens of Myxococcus xanthus cells undergoing submerged development.

    OpenAIRE

    Gill, J.S.; Dworkin, M

    1988-01-01

    Thirteen additional monoclonal antibodies directed against cell surface antigens of Myxococcus xanthus cells undergoing submerged development were isolated and partially characterized. As measured by quantitative enzyme-linked immunosorbent assay, 10 of these antibodies recognized antigens common to both vegetatively growing cells and cells undergoing submerged development; 3 antibodies recognized antigens specific to developing cells. Five antigens were revealed as single bands on Western bl...

  7. In silico design and performance of peptide microarrays for breast cancer tumour-auto-antibody testing

    Directory of Open Access Journals (Sweden)

    Andreas Weinhäusel

    2012-06-01

    Full Text Available The simplicity and potential of minimally invasive testing using sera from patients makes auto-antibody based biomarkers a very promising tool for use in cancer diagnostics. Protein microarrays have been used for the identification of such auto-antibody signatures. Because high throughput protein expression and purification is laborious, synthetic peptides might be a good alternative for microarray generation and multiplexed analyses. In this study, we designed 1185 antigenic peptides, deduced from proteins expressed by 642 cDNA expression clones found to be sero-reactive in both breast tumour patients and controls. The sero-reactive proteins and the corresponding peptides were used for the production of protein and peptide microarrays. Serum samples from females with benign and malignant breast tumours and healthy control sera (n=16 per group were then analysed. Correct classification of the serum samples on peptide microarrays were 78% for discrimination of ‘malignant versus healthy controls’, 72% for ‘benign versus malignant’ and 94% for ‘benign versus controls’. On protein arrays, correct classification for these contrasts was 69%, 59% and 59%, respectively. The over-representation analysis of the classifiers derived from class prediction showed enrichment of genes associated with ribosomes, spliceosomes, endocytosis and the pentose phosphate pathway. Sequence analyses of the peptides with the highest sero-reactivity demonstrated enrichment of the zinc-finger domain. Peptides’ sero-reactivities were found negatively correlated with hydrophobicity and positively correlated with positive charge, high inter-residue protein contact energies and a secondary structure propensity bias. This study hints at the possibility of using in silico designed antigenic peptide microarrays as an alternative to protein microarrays for the improvement of tumour auto-antibody based diagnostics.

  8. Development of Multiple ELISAs for the Detection of Antibodies against Classical Swine Fever Virus in Pig Sera

    Institute of Scientific and Technical Information of China (English)

    Zhen-hua Yang; Ling Li; Zi-shu Pan

    2012-01-01

    The major immunogenic proteins (Ems,E2 and NS3) of classical swine fever virus (CSFV) (Shimen strain) were expressed in E.coli and purified by affinity chromatography.The recombinant antigens were applied to develop multiple enzyme-linked immunosorbent assays (ELISAs) for the detection of specific antibodies in pig sera.Optimum cut-off values were determined by receiver operating characteristic (ROC) analysis after testing 201 sera of vaccinated pigs and 64 negative sera of unvaccinated piglets.The multiple ELISAs were validated with 265 pig sera yielding high sensitivity and specificity in comparison with the virus neutralization results.The results demonstrated that multiple ELISAs can be a valuable tool for the detection of CSFV infection and serological surveys in CSFV-free countries or for the evaluation of the antibody responses in pigs induced by a live attenuated C-strain vaccination.

  9. Production of antibody labeled gold nanoparticles for influenza virus H5N1 diagnosis kit development

    International Nuclear Information System (INIS)

    Preparation of colloidal gold conjugated antibodies specific for influenza A/H5N1 and its use in developing a virus A/H5N1 rapid diagnostic kit is presented. Colloidal gold nanoparticles (AuNPs) were prepared through citrate reduction. Single chain antibodies specific to H5N1 (scFv7 and scFv24) were produced using pTI2 + vector and E. coli strain HB2151. These antibodies were purified by affinity chromatography technique employing HiTrap Chelating HP columns pre-charged with Ni2 + . The method for preparation of antibody–colloidal gold conjugate was based on electrostatic force binding antibody with colloidal gold. The effect of factors such as pH and concentration of antibody has been quantitatively analyzed using spectroscopic methods after adding 1 wt% NaCl which induced AuNP aggregation. The morphological study by scanning electron microscopy (SEM) showed that the average size of the spherical AuNPs was 23 nm with uniform sizes. The spectroscopic properties of colloidal AuNPs showed the typical surface plasmon resonance band at 523 nm in UV-visible spectrum. The optimal pH of conjugated colloidal gold was found between 8.0 and 10.0. The activity of synthesized antibody labeled AuNPs for detection of H5N1 flu virus was checked by dot blot immunological method. The results confirmed the ability in detection of the A/H5N1 virus of the prepared antibody labeled gold particles and opened up the possibility of using them in manufacturing rapid detection kit for this virus. (paper)

  10. Detection of leptospiral antibodies by microscopic agglutination test in north-east of Iran

    Institute of Scientific and Technical Information of China (English)

    Ehsanollah Sakhaee; Gholam Reza Abdollah pour

    2011-01-01

    Objective: To detect leptospiral antibodies by microscopic agglutination test (MAT) in north-east of Iran. Methods: This study was conducted to evaluate prevalence of human leptospiral infections by MAT, using six current reference strains of Leptospira interrogans in north-east of Iran. A total of 285 serum samples were collected from three north-east provinces of Iran, from December, 2009 to June, 2010. Results: Antibodies were detected at least against one serovar of Leptospira interrogans in 45 sera (15.79 %) among 285 samples at a dilution 1:100 or greater. Positive titers against more than one serovar were detected in 24 sera of the positive samples. Therefore, there were 75 positive reactions against different serovar of Leptospira interrogans. Positive titers were recorded against serovar icterohaemorrhagiae (31 samples), hardjo (26 samples), grippotyphosa (7 samples), pomona (5 samples), canicola (4 samples) and ballum (2 sample).Conclusions:In present study the most prevalent (Leptospira icterohaemorrhagiae) and the least prevalent (Leptospira ballum) serovar are different from previous studies. Maybe, species and prevalence of serovars change during the time in one area and between regions.

  11. The Anti-Acetylcholine Receptor Antibody Test in Suspected Ocular Myasthenia Gravis

    Directory of Open Access Journals (Sweden)

    Jung Jin Lee

    2014-01-01

    Full Text Available Aim. To estimate the clinical significance of anti-acetylcholine receptor antibody (anti-AChR-Ab levels in suspected ocular myasthenia gravis. Methods. In total, 144 patients complaining of fluctuating diplopia and ptosis were evaluated for serum levels of anti-acetylcholine receptor antibody and their medical charts were retrospectively reviewed. Subjects were classified into three groups: variable diplopia only, ptosis only, and both variable diplopia and ptosis. We investigated serum anti-AChR-Ab titer levels and performed thyroid autoantibody tests. Results. Patients’ chief complaints were diplopia (N=103, ptosis (N=12, and their concurrence (N=29. Abnormal anti-AChR-Ab was observed in 21 of 144 patients (14.1%. Between the three groups, mean age, number of seropositive patients, and mean anti-AChR-Ab level were not significantly different (P=0.224, 0.073, and 0.062, resp.. Overall, 27.5% of patients had abnormal thyroid autoantibodies. Conclusion. The sensitivity of anti-AChR-Ab was 14.1% in suspected ocular myasthenia gravis and seropositivity in myasthenia gravis patients showed a high correlation with the presence of thyroid autoantibodies.

  12. Detector development and test facility

    International Nuclear Information System (INIS)

    Following the ideas presented in the proposal to the DoE, we have begun to acquire the equipment needed to design, develop construct and test the electronic and mechanical features of detectors used in High Energy Physics Experiments. A guiding principle for the effort is to achieve integrated electronic and mechanical designs which meet the demanding specifications of the modern hadron collider environment yet minimize costs. This requires state of the art simulation of signal processing as well as detailed calculations of heat transfer and finite element analysis of structural integrity

  13. Development of an enzyme-linked immunosorbent assay for detection of IgM antibodies to Babesia bigemina in cattle

    Directory of Open Access Journals (Sweden)

    Ruiz Patrícia M Gonçalves

    2001-01-01

    Full Text Available A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94% and sensitivity of the Elisa 87.5%. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina, or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days. The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina.

  14. Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples

    Institute of Scientific and Technical Information of China (English)

    WANG,Wen-Jun; LING,Yun; XU,Ting; GAO,Hong-Bin; SHENG,Wei; LI,Ji

    2007-01-01

    An indirect competitive enzyme-linked immunosorbent assay (icELISA) based on polyclonal antibody for the estrogen diethylstilbestrol (DES) was developed. With this aim, two different haptens mono-O-3-carboxypropyldiethylstilbestrol (DES-CP) and mono-O-carboxymethyldiethylstilbestrol (DES-CM) with carboxylic group that preserve the molecular structure character of diethylstilbestrol were synthesized. The haptens were conjugated with the carrier proteins bovine serum albumin (BSA) by mixed-anhydride method for immunogen and conjugated with ovalbumin (OVA) by active ester method for coating antigen. Polyclonal antibodies for diethylstilbestrol were raised by immunizing mice with immune antigen DES-CP-BSA. Under optimized system, the lowest limit of detection (LLD) of diethylstilbestrol was 0.01 ng/mL, and IC50= 1.02 ng/mL. Its analogs were tested and no obvious cross-reactivity was found to anti-diethylstilbestrol antibody. DES-fortified water samples were determined by simple dilution to diminish the matrix effect. The comparison between the amount of DES estimated by ELISA and the amount added indicates good agreement for all water samples tested, with mean recovery values ranging from 86% to 120.2%.

  15. Chlamydia antibody testing and diagnosing tubal pathology in subfertile women : an individual patient data meta-analysis

    NARCIS (Netherlands)

    Broeze, K. A.; Opmeer, B. C.; Coppus, S. F. P. J.; Van Geloven, N.; Alves, M. F. C.; Anestad, G.; Bhattacharya, S.; Allan, J.; Guerra-Infante, M. F.; Den Hartog, J. E.; Land, J. A.; Idahl, A.; Van der Linden, P. J. Q.; Mouton, J. W.; Ng, E. H. Y.; Van der Steeg, J. W.; Steures, P.; Svenstrup, H. F.; Tiitinen, A.; Toye, B.; Van der Veen, F.; Mol, B. W.

    2011-01-01

    BACKGROUND: The Chlamydia IgG antibody test (CAT) shows considerable variations in reported estimates of test accuracy, partly because of the use of different assays and cut-off values. The aim of this study was to reassess the accuracy of CAT in diagnosing tubal pathology by individual patient data

  16. Chlamydia antibody testing and diagnosing tubal pathology in subfertile women: an individual patient data meta-analysis

    NARCIS (Netherlands)

    Broeze, K.A.; Opmeer, B.C.; Coppus, S.F.; Geloven, N. van; Alves, M.F.; Anestad, G.; Bhattacharya, S.; Allan, J.; Guerra-Infante, M.F.; Hartog, J.E. Den; Land, J.A.; Idahl, A.; Linden, P.J. van der; Mouton, J.W.; Ng, E.H.; Steeg, J.W. van der; Steures, P.; Svenstrup, H.F.; Tiitinen, A.; Toye, B.; Veen, F. van der; Mol, B.W.

    2011-01-01

    BACKGROUND: The Chlamydia IgG antibody test (CAT) shows considerable variations in reported estimates of test accuracy, partly because of the use of different assays and cut-off values. The aim of this study was to reassess the accuracy of CAT in diagnosing tubal pathology by individual patient data

  17. Development of a serotype colloidal gold strip using monoclonal antibody for rapid detection type Asia1 foot-and-mouth disease

    OpenAIRE

    Gao Shan-dian; Cong Guo-zheng; Du Jun-zheng; Shao Jun-jun; Lin Tong; Chang Huiyun

    2011-01-01

    Abstract Background In this study, we developed a rapid, one step colloid gold strip (CGS) capable of specifically detecting type Asia1 foot-and-mouth disease virus (FMDV). We have produced two monoclonal antibodies (mAb) to type Asia1 FMD (named 1B8 and 5E2). On the test strip, the purified 1B8 labelled with the colloidal gold was used as the detector, and the purified 5E2 and goat anti-mouse antibodies were wrapped onto nitrocellulose (NC) membranes as the test and the control line, respect...

  18. Clinical development methodology for infusion-related reactions with monoclonal antibodies.

    Science.gov (United States)

    Doessegger, Lucette; Banholzer, Maria Longauer

    2015-07-01

    Infusion-related reactions (IRRs) are common with monoclonal antibodies (mAbs) and timely related to drug administration and have been reported as anaphylaxis, anaphylactoid reactions and cytokine release syndrome, among other terms used. We address risk management measures for individual patients and for the study and propose a consistent reporting approach in an attempt to allow cross-molecule comparisons. Once the symptoms of IRR have resolved, the mAb may be restarted. Rechallenge should not be done for suspected IgE-mediated anaphylaxis and Grade 4 IRRs. Management of IRRs for subsequent patients includes administration of premedication, which, however, does not prevent IgE-mediated anaphylaxis. Reporting approach: (1) Report as IRRs, reactions occurring during or within 24 h after an infusion. Negative skin Prick test and absent or undetectable allergen-specific IgE levels have high negative predictive value for an IgE-mediated allergic reaction. If IgE-mediated anaphylaxis is suspected based on medical history and/or laboratory test results, the reaction should be reported as suspected (IgE mediated) anaphylaxis. (2) Collect signs and symptoms with grades to allow characterization of IRRs. IRRs pathogenesis is of scientific interest and has impact on drug development. Animal toxicology studies are neither predictive of severe IRRs nor of anaphylaxis in human. Preclinical tests should be further developed to identify patients at risk for severe IRRs, for complement activation-related pseudoallergy and for IgE-mediated anaphylaxis. The proposed approach should help standardizing data collection and analysis of IRRs in an attempt to enable comparisons across molecules. PMID:26246897

  19. Alternative Water Processor Test Development

    Science.gov (United States)

    Pickering, Karen D.; Mitchell, Julie; Vega, Leticia; Adam, Niklas; Flynn, Michael; Wjee (er. Rau); Lunn, Griffin; Jackson, Andrew

    2012-01-01

    The Next Generation Life Support Project is developing an Alternative Water Processor (AWP) as a candidate water recovery system for long duration exploration missions. The AWP consists of biological water processor (BWP) integrated with a forward osmosis secondary treatment system (FOST). The basis of the BWP is a membrane aerated biological reactor (MABR), developed in concert with Texas Tech University. Bacteria located within the MABR metabolize organic material in wastewater, converting approximately 90% of the total organic carbon to carbon dioxide. In addition, bacteria convert a portion of the ammonia-nitrogen present in the wastewater to nitrogen gas, through a combination of nitrogen and denitrification. The effluent from the BWP system is low in organic contaminants, but high in total dissolved solids. The FOST system, integrated downstream of the BWP, removes dissolved solids through a combination of concentration-driven forward osmosis and pressure driven reverse osmosis. The integrated system is expected to produce water with a total organic carbon less than 50 mg/l and dissolved solids that meet potable water requirements for spaceflight. This paper describes the test definition, the design of the BWP and FOST subsystems, and plans for integrated testing.

  20. Alternative Water Processor Test Development

    Science.gov (United States)

    Pickering, Karen D.; Mitchell, Julie L.; Adam, Niklas M.; Barta, Daniel; Meyer, Caitlin E.; Pensinger, Stuart; Vega, Leticia M.; Callahan, Michael R.; Flynn, Michael; Wheeler, Ray; Birmele, Michele; Lunn, Griffin; Jackson, Andrew

    2013-01-01

    The Next Generation Life Support Project is developing an Alternative Water Processor (AWP) as a candidate water recovery system for long duration exploration missions. The AWP consists of biological water processor (BWP) integrated with a forward osmosis secondary treatment system (FOST). The basis of the BWP is a membrane aerated biological reactor (MABR), developed in concert with Texas Tech University. Bacteria located within the MABR metabolize organic material in wastewater, converting approximately 90% of the total organic carbon to carbon dioxide. In addition, bacteria convert a portion of the ammonia-nitrogen present in the wastewater to nitrogen gas, through a combination of nitrification and denitrification. The effluent from the BWP system is low in organic contaminants, but high in total dissolved solids. The FOST system, integrated downstream of the BWP, removes dissolved solids through a combination of concentration-driven forward osmosis and pressure driven reverse osmosis. The integrated system is expected to produce water with a total organic carbon less than 50 mg/l and dissolved solids that meet potable water requirements for spaceflight. This paper describes the test definition, the design of the BWP and FOST subsystems, and plans for integrated testing.

  1. Development of novel monoclonal antibodies that define differentiation stages of human stromal (mesenchymal) stem cells

    DEFF Research Database (Denmark)

    Andersen, Ditte Caroline; Kortesidis, Angela; Zannettino, Andrew C W;

    2011-01-01

    Human mesenchymal stem cells (hMSC) are currently being introduced for cell therapy, yet, antibodies specific for native and differentiated MSCs are required for their identification prior to clinical use. Herein, high quality antibodies against MSC surface proteins were developed by immunizing...... differentiation. Interestingly, undifferentiated cells revealed a sole cytoplasmic distribution pattern of Collagen VI, which however changed to an extracellular matrix appearance upon osteogenic- and adipogenic differentiation. In relation to this, we found that STRO-1(+/-)/Collagen VI(-) sorted hMSC contained...... fewer differentiated alkaline phosphatase(+) cells compared to STRO-1(+/-)/Collagen VI(+) hMSC, suggesting that Collagen VI on the cell membrane exclusively defines differentiated MSCs. In conclusion, we have generated a panel of high quality antibodies to be used for characterization of MSCs, and in...

  2. Development and Characterization of Mouse Monoclonal Antibodies Reactive with Chicken CD83

    Science.gov (United States)

    This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein con...

  3. Development of a monoclonal antibody against human heart ferritin and its application in an immunoradiometric assay

    Energy Technology Data Exchange (ETDEWEB)

    Cavanna, F.; Chieregatti, G.; Murador, E. (Farmitalia Carlo Erba, Milano (Italy). Ricerche e Sviluppo Diagnostici); Ruggeri, G.; Iacobello, C.; Albertini, A. (Facolta di Medicina, Brescia (Italy). Cattedra di Chimica); Arosio, P. (Milan Univ. (Italy). Dip. di Scienze e Tecnologie Biomediche)

    1983-11-15

    In the present report the authors describe the development of a monoclonal antibody which appears to be strictly monospecific for acidic human isoferritins. Its characteristics of specificity and affinity have been studied, and it is shown that it can be used in an immunoradiometric assay to evaluate the acidic ferritin level in serum.

  4. 78 FR 7438 - Prospective Grant of Exclusive License: Development of Human Monoclonal Antibodies Against DR4

    Science.gov (United States)

    2013-02-01

    ... HUMAN SERVICES National Institutes of Health Prospective Grant of Exclusive License: Development of... America. The prospective start-up exclusive commercial license territory may be worldwide and the field of... antibodies could be used as a research tool for the study of DR4. The prospective start-up...

  5. Development and Use of Fluorescent Antibody and qPCR Protocols for the Electrostatic Spore Trap

    Science.gov (United States)

    Fluorescent antibody (FA) and qPCR protocols were evaluated for the newly developed aerobiological sampler (Ionic Spore Trap), which depends upon electrostatic deposition of particulates onto a 25 mm aluminum disk (stub). This device was originally designed for assessment of captured particulates by...

  6. Suitability of a Rapid Immunochromatographic Test for Detection of Antibodies to Human Immunodeficiency Virus in Ghana, West Africa

    OpenAIRE

    Aidoo, S.; Ampofo, W. K.; Brandful, J. A. M.; Nuvor, S. V.; Ansah, J. K.; Nii-Trebi, N.; Barnor, J. S.; Apeagyei, F; Sata, T.; Ofori-Adjei, D.; Ishikawa, K.

    2001-01-01

    In West African countries such as Ghana, efficient human immunodeficiency virus (HIV) testing is a priority in the fight against AIDS. A new immunochromatographic rapid test, Determine HIV-1/2 (Abbott Diagnostics, North Chicago, Ill.), that detects antibodies against HIV type 1 (HIV-1) and/or HIV-2 was evaluated using Ghanaian blood samples. Two hundred four serum and/or plasma specimens were tested. HIV screening was done by a particle agglutination test and confirmed by a Western blot (WB) ...

  7. Development of a double-monoclonal antibody sandwich ELISA: Tool for chicken interferon-γ detection ex vivo.

    Science.gov (United States)

    Dai, Hua; Xu, Zheng-Zhong; Wang, Meiling; Chen, Jun-Hua; Chen, Xiang; Pan, Zhi-Ming; Jiao, Xin-An

    2016-04-01

    The aim of the present work was to develop reagents to set up a chicken interferon-γ (ChIFN-γ) assay. Four monoclonal antibodies (mAbs) specific for ChIFN-γ were generated to establish sandwich ELISA based on 2 different mAbs. To improve the detection sensitivity of ChIFN-γ, a double-monoclonal antibody sandwich ELISA was developed using mAb 3E5 as capture antibody and biotinylated mAb 3E3 as a detection reagent. The results revealed that this ELISA has high sensitivity, allowing for the detection of 125 to 500 pg/mL of recombinant ChIFN-γ, and also has an excellent capacity for detecting native ChIFN-γ. This ELISA was then used to detect ChIFN-γ level in chickens immunized with a Newcastle disease virus (NDV) vaccine, the immunized chicken splenocytes were stimulated by NDV F protein as recall antigen. From our results, it appears that the sensitivity range of this sandwich ELISA test is adequate to measure the ex vivo release of ChIFN-γ. PMID:27127340

  8. Cross-reactive neutralizing antibody responses to enterovirus 71 infections in young children: implications for vaccine development.

    Directory of Open Access Journals (Sweden)

    Mei-Liang Huang

    Full Text Available BACKGROUND: Recently, enterovirus 71 (EV71 has caused life-threatening outbreaks involving neurological and cardiopulmonary complications in Asian children with unknown mechanism. EV71 has one single serotype but can be phylogenetically classified into 3 main genogroups (A, B and C and 11 genotypes (A, B1∼B5 and C1∼C5. In Taiwan, nationwide EV71 epidemics with different predominant genotypes occurred in 1998 (C2, 2000-2001 (B4, 2004-2005 (C4, and 2008 (B5. In this study, sera were collected to measure cross-reactive neutralizing antibody titers against different genotypes. METHODS: We collected historical sera from children who developed an EV71 infection in 1998, 2000, 2005, 2008, or 2010 and measured cross-reactive neutralizing antibody titers against all 11 EV71 genotypes. In addition, we aligned and compared the amino acid sequences of P1 proteins of the tested viruses. RESULTS: Serology data showed that children infected with genogroups B and C consistently have lower neutralizing antibody titers against genogroup A (>4-fold difference. The sequence comparisons revealed that five amino acid signatures (N143D in VP2; K18R, H116Y, D167E, and S275A in VP1 are specific for genogroup A and may be related to the observed antigenic variations. CONCLUSIONS: This study documented antigenic variations among different EV71 genogroups and identified potential immunodominant amino acid positions. Enterovirus surveillance and vaccine development should monitor these positions.

  9. Development of a robust reporter-based assay for the bioactivity determination of anti-VEGF therapeutic antibodies.

    Science.gov (United States)

    Wang, Lan; Xu, Gang-Ling; Gao, Kai; Wilkinson, Jennifer; Zhang, Feng; Yu, Lei; Liu, Chun-Yu; Yu, Chuan-Fei; Wang, Wen-Bo; Li, Meng; Chen, Wei; Fan, Frank; Cong, Mei; Wang, Jun-Zhi

    2016-06-01

    Development of anti-VEGF based biologic agents has been a focus in cancer treatment for the past decades, and several anti-VEGF pharmaceuticals have been already approved for treatment of various medical indications especially in cancer. The first anti-angiogenic agent approved by FDA was bevacizumab (BVZ, trade name Avastin, Genentech/Roche), a humanized anti-VEGF monoclonal antibody. Accurate determination of bioactivity is crucial for the safety and efficacy of therapeutic antibodies. The current method widely used in the lot release and stability test for clinical trial batches of BVZ is anti-proliferation assay using primary human umbilical vein endothelial cells (HUVEC), which is tedious with high assay variations. We describe here the development and preliminary validation of a reporter gene assay (RGA) that is based on an HEK293 cell line stably expressing vascular endothelial growth factor receptor 2 (VEGFR-2), and a luciferase reporter under the control of nuclear factor activated T cell (NFAT) response elements. Our study shows this assay not only to be superior on precision, sensitivity and assay simplicity compared with HUVEC assay, but also applicable to other VEGF-targeted biotherapeutics. These results show for the first time that this new reporter assay, based on the VEGF-VEGFR-NFAT pathway, can be a viable supplement to the HUVEC assay and employed in potency determination of BVZ and other kinds of anti-VEGF antibody-based biotherapeutics. PMID:27042807

  10. Seroprevalence of bovine leptospiral antibodies by microscopic agglutination test in Southeast of Iran

    Institute of Scientific and Technical Information of China (English)

    Mohammad Khalili; Ehsanollah Sakhaee; Mohammad Reza Aflatoonian; Gholamreza Abdollahpour; Saeed Sattari Tabrizi; Elham Mohammadi Damaneh; Sajad Hossini-nasab

    2014-01-01

    Objective:To evaluate serological findings of bovine leptospirosis which is a zoonotic disease with worldwide distribution caused by Leptospira interrogans. Methods: One hundred and sixty seven sera were collected from 9 commercial dairy herds in jiroft suburbs, from July to October 2011. Microscopic agglutination test (MAT) was used to evaluates serological findings of bovine leptospirosis in Jiroft suburb dairy farms, Kerman province, Iran. Results:Antibodies were found by MAT at least against one serovar of Leptospira interrogans in 29 samples (17.36%) among 167 sera at a dilution 1:100 or higher, and Leptospira pomona was the most prevalent serovar. Positive titers against more than one serovar were detected in 6 sera of the positive samples. Conclusion:This study is the first report of leptospirosis in Southeast Iran and showed that Leptospira pomona was the most and Leptospira icterohaemorrhagiae the least prevalent serovars in Southeast Iran.

  11. Standardization of serological tests for detecting anti-Trypanosoma cruzi antibodies in dogs

    Directory of Open Access Journals (Sweden)

    M. A. Lauricella

    1993-09-01

    Full Text Available This paper reports on the standardization of four serological reactions currently used in human serodiagnosis for the detection of anti-Trypanosoma cruzi antibodies in naturally and experimentally infected dogs. Indirect immunofluorescence test (IFAT and hemagglutination test (IHAT were standardized, and complement fixation test (CFT and direct agglutination test (DAT were used for diagnostic confirmation. Four hundred and eighty one mongrel dogs that were studied by xenodiagnosis were used: (1 parasitemic dogs of two localities of endemic area (EA of Santiago del Estero province in Argentina (n = 134; (2 non-parasitemic dogs of the same area (n = 285; (3 dogs experimentally infected with T. cruzi in the patent period (n = 6; (4 non-infected dogs (n = 56 which were born in the city of Buenos Aires (BA, one non-EA for Chagas' disease. For IFAT, parasitemic dogs EA showed 95% of reactive sera. Non parasitemic dogs EA showed 77% of non reactive sera. None sera from BA were reactive for dilutions higher than four. For IHAT, 84% of sera of parasitemic dogs EA showed serological reactivity and among non parasitemic dogs BA, 61% were non reactive, while the remainder showed at most titres of 1/16. The cut-off titres for IFAT and IHAT were 1/16 and 1/32 respectively, and for CFT and DAT 1/1 and 1/128 respectively. Sensitivity for IFAT, IHAT, CF and DAT were 95%, 84%, 97% and 95% respectively.

  12. Detection of Rocky Mountain spotted fever antibodies by a latex agglutination test.

    OpenAIRE

    Hechemy, K E; Anacker, R L; Philip, R N; Kleeman, K T; MacCormack, J. N.; Sasowski, S J; Michaelson, E E

    1980-01-01

    A latex test for immunodiagnosis of Rocky Mountain spotted fever, using erythrocyte-sensitizing substance from Rickettsia rickettsii adsorbed to latex particles, has been developed. The test was evaluated with a total of 123 single and 118 paired human sera submitted for Rocky Mount spotted fever testing. This test is simple, sensitive, and specific. Its efficiency, relative to the reference microimmunofluorescence test, was 95.1% for single sera and approached 100% for paired sera.

  13. Effect of using heat-inactivated serum with the Abbott human T-cell lymphotropic virus type III antibody test.

    OpenAIRE

    Jungkind, D. L.; DiRenzo, S A; Young, S J

    1986-01-01

    The Abbott enzyme immunoassay (Abbott Laboratories, North Chicago, Ill.) for human T-cell lymphotropic virus type III (HTLV-III) antibody was evaluated to determine the effect of using heat-inactivated (56 degrees C for 30 min) serum as the sample. Each of 58 nonreactive serum samples gave a higher A492 value when tested after heat inactivation. Ten of the samples became reactive after heating. Heat-inactivated serum should not be used in the current Abbott HTLV-III antibody test, because thi...

  14. Attitudes towards HIV-antibody testing and people with aids among university students in India, South Africa and United States

    Directory of Open Access Journals (Sweden)

    Peltzer Karl

    2004-03-01

    Full Text Available CONTEXT: Stigmatizing attitudes toward persons with AIDS (PWAs may reduce people′s willingness to have themselves tested for Human Immunodeficiency Virus (HIV - thereby increasing the risk of transmission. AIMS: To examine attitudes towards (HIV testing and determinants of attitudes towards PWAs. SETTINGS AND DESIGN: A cross-sectional. MATERIAL AND METHODS: 600 first-year university students from South India, South African and America filled in a self-administered questionnaire. Main outcome measures included an Attitudes towards HIV-Antibody Testing Scale and Readiness to engage in personal forms of contact with People With AIDS. RESULTS indicate that the majority of American and South African students and only 10 percent of the Indian students had been sexually active in the past 12 months. Almost one fifth of the American and South African participants but only 10% of the Indian students admitted to having had an HIV test. American students had a much more positive attitudes toward HIV testing than South African and Indian students. Regression analysis for the Indian student sample identified blaming, irritation and negative attitudes toward homosexuals as independent predictors of readiness to engage in personal contact with PWAs, while the regression analyses for both South African and American students identified pity and irritation as independent predictors of contact readiness with PWAs. Positive HIV testing attitudes were positively associated with contact readiness with PWAs. CONCLUSION: The findings are important for the role of HIV testing and counselling in campus AIDS programmes. The findings reveal important factors related to HIV testing and suggest strategies for developing effective HIV/AIDS counselling programmes in universities.

  15. Harnessing T cells to fight cancer with BiTE(®) antibody constructs - past developments and future directions.

    Science.gov (United States)

    Klinger, Matthias; Benjamin, Jonathan; Kischel, Roman; Stienen, Sabine; Zugmaier, Gerhard

    2016-03-01

    Bispecific T-cell engager (BiTE(®) ) antibody constructs represent a novel immunotherapy that bridges cytotoxic T cells to tumor cells, thereby inducing target cell-dependent polyclonal T-cell activation and proliferation, and leading to apoptosis of bound tumor cells. Anti-CD19 BiTE(®) blinatumomab has demonstrated clinical activity in Philadelphia chromosome (Ph)-negative relapsed or refractory (r/r) acute lymphoblastic leukemia (ALL) eventually resulting in conditional approval by the U.S. Food and Drug Administration in 2014. This drug is currently further developed in pediatric and Ph(+) r/r, as well as in minimal residual disease-positive ALL, and might also offer clinical benefit for patients with non-Hodgkin's lymphoma, especially for those with aggressive forms like diffuse large B-cell lymphoma. Another BiTE(®) antibody construct in hemato-oncology designated AMG 330 targets CD33 on acute myeloid leukemia blast cells. After showing promising ex vivo activity, this drug candidate has recently entered phase 1 clinical development, and has further indicated potential for combination with checkpoint inhibitors. In solid tumor indications, three BiTE(®) antibody constructs have been tested in phase 1 studies so far: anti-EpCAM BiTE(®) AMG 110, anti-CEA BiTE(®) MEDI-565/AMG 211, and anti-PSMA BiTE(®) BAY2010112/AMG 212. Pertinent questions comprise how to maximize BiTE(®) penetration and T-cell infiltration of the tumor while simultaneously minimizing any adverse events, which is currently explored by a continuous intravenous infusion approach. Thus, BiTE(®) antibody constructs will hopefully provide new treatment options for patients in several indications with high unmet medical need. PMID:26864113

  16. Antibody recognition of the dengue virus proteome and implications for development of vaccines.

    Science.gov (United States)

    Fernandez, Stefan; Cisney, Emily D; Tikhonov, Alexander P; Schweitzer, Barry; Putnak, Robert J; Simmons, Monika; Ulrich, Robert G

    2011-04-01

    Dengue is a mosquito-borne infection caused by four distinct serotypes of dengue virus, each appearing cyclically in the tropics and subtropics along the equator. Although vaccines are currently under development, none are available to the general population. One of the main impediments to the successful advancement of these vaccines is the lack of well-defined immune correlates of protection. Here, we describe a protein microarray approach for measuring antibody responses to the complete viral proteome comprised of the structural (capsid, membrane, and envelope) and nonstructural (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) components of all four dengue virus serotypes (1 to 4). We examined rhesus macaques vaccinated with tetravalent vaccines consisting of live-attenuated virus (LAV) or purified inactivated virus (PIV), followed by boosting with LAV and challenging with wild-type dengue virus. We detected temporal increases in antibodies against envelope proteins in response to either vaccine, while only the PIV/LAV vaccination strategy resulted in anticapsid antibodies. In contrast to results from vaccination, naïve macaques challenged with wild-type viruses of each serotype demonstrated a balanced response to nonstructural and structural components, including responses against the membrane protein. Our results demonstrate discriminating details concerning the nature of antibody responses to dengue virus at the proteomic level and suggest the usefulness of this information for vaccine development. PMID:21270280

  17. Cellular impedance measurement as a new tool for poxvirus titration, antibody neutralization testing and evaluation of antiviral substances

    International Nuclear Information System (INIS)

    Research highlights: → Real-time data acquisition by RT-CES requires low operative effort. → Time to result is reduced by using RT-CES instead of conventional methods. → RT-CES enables quantification of virus titers in unknown samples. → RT-CES is a useful tool for high-throughput characterization of antiviral agents. → An RT-CES-based virus neutralization test was established. -- Abstract: Impedance-based biosensing known as real-time cell electronic sensing (RT-CES) belongs to an emerging technology for analyzing the status of cells in vitro. In the present study protocols were developed for an RT-CES-based system (xCELLigenceTM, Roche Applied Science, ACEA Biosciences Inc.) to supplement conventional techniques in pox virology. First, proliferation of cells susceptible to orthopoxviruses was monitored. For virus titration cells were infected with vaccinia virus and cell status, represented by the dimensionless impedance-based cell index (CI), was monitored. A virus-dose dependent decrease in electrical impedance could be shown. Calculation of calibration curves at a suitable CI covering a dynamic range of 4 log enabled the quantification of virus titers in unknown samples. Similarly, antiviral effects could be determined as shown for anti-poxviral agents ST-246 and Cidofovir. Published values for the in vitro concentration that inhibited virus replication by 50% (IC50) could be confirmed while cytotoxicity in effective concentrations was excluded in long-term incubation experiments. Finally, an RT-CES-based virus neutralization test was established. Various poxvirus-specific antibodies were examined for their neutralizing activity and a calculation mode for the neutralizing antibody titer was introduced. In summary, the presented RT-CES-based methods outmatch end-point assays by observing the cell population throughout the entire experiment while workload and time to result are reduced.

  18. Cellular impedance measurement as a new tool for poxvirus titration, antibody neutralization testing and evaluation of antiviral substances

    Energy Technology Data Exchange (ETDEWEB)

    Witkowski, Peter T. [Robert Koch-Institut, Zentrum fuer Biologische Sicherheit 1, Nordufer 20, 13353 Berlin (Germany); Charite Universitaetsmedizin, CCM, Institut fuer Virologie, Helmut Ruska Haus, Chariteplatz 1, 10117 Berlin (Germany); Schuenadel, Livia, E-mail: SchuenadelL@rki.de [FU-Berlin, Fachbereich Biologie, Chemie, Pharmazie, Takustrasse 3, 14195 Berlin (Germany); Robert Koch-Institut, Zentrum fuer Biologische Sicherheit 1, Nordufer 20, 13353 Berlin (Germany); Wiethaus, Julia; Bourquain, Daniel R.; Kurth, Andreas; Nitsche, Andreas [Robert Koch-Institut, Zentrum fuer Biologische Sicherheit 1, Nordufer 20, 13353 Berlin (Germany)

    2010-10-08

    Research highlights: {yields} Real-time data acquisition by RT-CES requires low operative effort. {yields} Time to result is reduced by using RT-CES instead of conventional methods. {yields} RT-CES enables quantification of virus titers in unknown samples. {yields} RT-CES is a useful tool for high-throughput characterization of antiviral agents. {yields} An RT-CES-based virus neutralization test was established. -- Abstract: Impedance-based biosensing known as real-time cell electronic sensing (RT-CES) belongs to an emerging technology for analyzing the status of cells in vitro. In the present study protocols were developed for an RT-CES-based system (xCELLigence{sup TM}, Roche Applied Science, ACEA Biosciences Inc.) to supplement conventional techniques in pox virology. First, proliferation of cells susceptible to orthopoxviruses was monitored. For virus titration cells were infected with vaccinia virus and cell status, represented by the dimensionless impedance-based cell index (CI), was monitored. A virus-dose dependent decrease in electrical impedance could be shown. Calculation of calibration curves at a suitable CI covering a dynamic range of 4 log enabled the quantification of virus titers in unknown samples. Similarly, antiviral effects could be determined as shown for anti-poxviral agents ST-246 and Cidofovir. Published values for the in vitro concentration that inhibited virus replication by 50% (IC{sub 50}) could be confirmed while cytotoxicity in effective concentrations was excluded in long-term incubation experiments. Finally, an RT-CES-based virus neutralization test was established. Various poxvirus-specific antibodies were examined for their neutralizing activity and a calculation mode for the neutralizing antibody titer was introduced. In summary, the presented RT-CES-based methods outmatch end-point assays by observing the cell population throughout the entire experiment while workload and time to result are reduced.

  19. Evaluation of an In-House-Developed Radioassay Kit for Antibody Detection in Cases of Pulmonary Tuberculosis and Tuberculous Meningitis

    OpenAIRE

    Kameswaran, M.; Shetty, K.; Ray, M. K.; Jaleel, M. A.; Kadival, G. V.

    2002-01-01

    A radioassay for the detection of antitubercular antibody has been developed. The technique involves the addition of 125I-labeled Mycobacterium tuberculosis antigen as a tracer, diluted clinical sample (serum or cerebrospinal fluid [CSF]), and heat-inactivated Staphylococcus aureus to capture the antibody, incubation for 4 h, and quantitation of the amount of antibody present in the sample. A total of 330 serum samples from patients with pulmonary tuberculosis and 138 control serum samples fr...

  20. Radionuclide antibody-conjugates: developments and applications to obtain a targeted cancer therapy

    OpenAIRE

    Gjorgieva Ackova, Darinka; Smilkov, Katarina; Makreski, Petre; Stafilov, Trajče; Duatti, Adriano; Janevik-Ivanovska, Emilija

    2015-01-01

    Understanding the behaviour and function of biomolecules at the molecular level is key to the discovery and development of new drugs, as well as diagnostic techniques. The characterization of therapeutic monoclonal antibodies (mAbs) poses many challenges compared to those of low-molecular mass drugs because of their inherent complexity due to their protein nature. Achievements in this field of science have changed the way that drugs are being designed and developed nowadays. Vibrational spect...

  1. Clinical relevance of testing for antineutrophil cytoplasm antibodies (ANCA) with a standard indirect immunofluorescence ANCA test in patients with upper or lower respiratory tract symptoms.

    OpenAIRE

    Davenport, A; Lock, R J; Wallington, T B

    1994-01-01

    BACKGROUND--Reports from specialist nephrological centres have suggested that the antineutrophil cytoplasm antibody (ANCA) test is highly specific and sensitive for patients with Wegener's granulomatosis. To determine the usefulness of the ANCA test in everyday respiratory practice the results of the test were audited in all patients in the south west of England with respiratory symptoms who underwent the test. METHODS--The results of all 335 patients who had presented with upper or lower res...

  2. Test driven software development with Java EE

    OpenAIRE

    Balantič, Matija

    2013-01-01

    Test driven development (TDD) is a technique with the main idea of writing a failing test first, which is then made to pass by implementing a particular snippet of code. Development is done in short iterations which consist of three basic steps, namely test-code-refactor. The thesis shows the development of Java EE web applications FerApp using test driven development and continuous integration. The application development was driven with unit tests and complemented with integration and f...

  3. Lowering the cut off value of an automated chlamydia enzyme immunoassay and confirmation by PCR and direct immunofluorescent antibody test.

    OpenAIRE

    Tong, C Y; Donnelly, C; Hood, N

    1997-01-01

    AIMS: To increase the sensitivity of an automated chlamydia enzyme immunoassay by significantly lowering its cut off value, and to maintain specificity by confirmation with polymerase chain reaction (PCR) and direct immunofluorescent antibody test (DFA). METHODS: Over five months, the cut off value of the enzyme immunoassay used to screen urogenital samples for chlamydia antigen was reduced from 80 to 10. Samples with a test value of 10 or above were further tested with a commercial PCR assay...

  4. Monoclonal antibodies

    International Nuclear Information System (INIS)

    Monoclonal antibodies (MAbs) are antibodies having single specificity for a given antigen site (epitope). The development of hybridoma technology and the relative ease by which MAbs can be prepared has revolutionized many aspects of serological applications in diagnosis and differentiation of disease producing agents. The property of monospecificity offers advantages in diagnostic applications over polyclonal sera in that tests can be defined exactly with regard to the antigen detected and the affinity of reaction between the given antigenic site and the monoclonal reagent. In addition, MAbs offer better possibilities for test standardization, because the same reagent can be used in different laboratories. Such an MAb can be supplied by a central laboratory or 'grown' from hybridoma cells, ensuring that the resultant product is identical from laboratory to laboratory and that the part of the test involving the MAb reaction is the same. The methodologies for inoculation regimes, mice, cloning methods, selection of fusion partners, etc., have been validated extensively in developed country laboratories. The decision to establish a MAb production facility must be examined on a strict cost-benefit basis, since it is still expensive to produce a product. There are many MAbs available that should be sought to allow exploitation in developing tests. If a production facility is envisaged, it should produce reagents for national needs, i.e. there should be a clear problem oriented approach whereby exact needs are defined. In the field of veterinary applications, MAbs are the central reagent in many immunoassays based on the enzyme linked immunosorbent assay (ELISA). The development of specific tests for diagnosing diseases is dominated by MAbs and has been fuelled by a strong research base, mainly in developed countries allied to developing countries through the study of related diseases. Thus, there are very many assays dependent on MAbs, some of which form the basis of

  5. Chikungunya virus RNA and antibody testing at a National Reference Laboratory since the emergence of Chikungunya virus in the Americas.

    Science.gov (United States)

    Prince, Harry E; Seaton, Brent L; Matud, Jose L; Batterman, Hollis J

    2015-03-01

    Since first reported in the Americas in December 2013, chikungunya virus (CHIKV) infections have been documented in travelers returning from the Caribbean, with many cases identified by CHIKV antibody and/or RNA testing at our laboratory. We used our large data set to characterize the relationship between antibody titers and RNA detection and to estimate IgM persistence. CHIKV RNA was measured by nucleic acid amplification and CHIKV IgG/IgM by indirect immunofluorescence. Of the 1,306 samples submitted for RNA testing in January through September 2014, 393 (30%) were positive; for 166 RNA-positive samples, CHIKV antibody testing was also ordered, and 84% were antibody negative. Of the 6,971 sera submitted for antibody testing in January through September 2014, 1,811 (26%) were IgM positive; 1,461 IgM positives (81%) were also IgG positive. The relationship between the CHIKV antibody titers and RNA detection was evaluated using 376 IgM-positive samples (138 with RNA testing ordered and 238 deidentified and tested for RNA). RNA detection showed no significant association with the IgM titer but was inversely related to the IgG titer; 63% of the IgG negative sera were RNA positive, compared to 36% of sera with low IgG titers (1:10 to 1:80) and 16% with IgG titers of ≥1:160. Using second-sample results from 62 seroconverters, we estimated that CHIKV IgM persists for 110 days (95% confidence interval, 78 to 150 days) after the initial antibody-negative sample. These findings indicate that (i) RNA detection is more sensitive than antibody detection early in CHIKV infection, (ii) in the absence of RNA results, the IgG titer of the IgM-positive samples may be a useful surrogate for viremia, and (iii) CHIKV IgM persists for approximately 4 months after symptom onset. PMID:25540275

  6. Mollusc reproductive toxicity tests - Development and validation of test guidelines

    DEFF Research Database (Denmark)

    Ducrot, Virginie; Holbech, Henrik; Kinnberg, Karin Lund

    ). Cumulated fecundity per individual over the test period was SETAC 6th World Congress/SETAC Europe 22nd Annual Meeting 223 the main test endpoint. Tested animals came from a single laboratory culture (i.e. the University of Frankfurt for P. antipodarum and INRA for L. stagnalis). Snails were acclimated in...... experimental conditions and test procedures is required before the SOPs are ready to be proposed as OECD test guidelines.......The Organisation for Economic Cooperation and Development is promoting the development and validation of mollusc toxicity tests within its test guidelines programme, eventually aiming for the standardization of mollusc apical toxicity tests. Through collaborative work between academia, industry and...

  7. Comparison of the 2-mercaptoethanol and dithiothreitol tests for determining Brucella immunoglobulin G agglutinating antibody in bovine serum.

    OpenAIRE

    McMahon, K. J.

    1983-01-01

    The dithiothreitol test was evaluated as a substitute for the 2-mercaptoethanol test for determining Brucella immunoglobulin G agglutinating antibody in bovine serum. The tests were compared on 207 card-positive sera that showed a standard tube-agglutination titer of incomplete 1:50 or higher. The tests agreed within one dilution with 182 of the 207 sera tested for an 87.9% rate of agreement. When titers were not the same, those obtained with the dithiothreitol test were more frequently lower...

  8. Development of a protein biochip to identify 6 monoclonal antibodies against subtypes of recombinant human interferons.

    Science.gov (United States)

    Xu, Zhenshan; Du, Weidong; Zhang, Ping; Wang, Xuan; Ma, Xueling; Shi, Liqin; Song, Lihua

    2010-04-01

    Recombinant human interferons (rhIFNs) are broadly used as effective therapeutic agents with antiviral, antitumor, and immune-modulating properties. Advances in protein biochip technology have benefited the medical community greatly, making true parallelism, miniaturization, and high throughput possible. In this study, 5 rhIFN proteins (IFN-alpha1b, IFN-alpha2a, IFN-alpha2b, IFN-beta, and IFN-gamma) were immobilized onto an N-hydroxysuccinimide (NHS)-modified gold-based biochip. The protein biochip was incubated with 6 specific mouse IgG antibodies (AK1, AK2, AK3, AK4, BK1, and CK1) against the human IFNs and then with Cy3-conjugated goat anti-mouse IgG antibody. The results showed that monoclonal antibody AK1 presented a unique binding characteristic to IFN-alpha1b. AK2 reacted in immunoassays equally with IFN-alpha2a and IFN-alpha2b. AK3 detected IFN-alpha1b, IFN-alpha2a, and IFN-alpha2b. AK4 had positive immunological responses directed to both IFN-alpha1b and IFN-alpha2b. Monoclonal antibodies BK1 and CK1 recognized epitope of IFN-beta and IFN-gamma, specifically. The assay specificity of the biochip was further confirmed by enzyme-linked immunosorbent assay (ELISA) and western blotting. Finally, 88 serum samples from patients treated with rhIFN-alpha2b were simultaneously tested on a single biochip. The result demonstrated that 6.8% (6 of 88 cases) presented positive reactions to anti-IFN-alpha2b antibodies, indicating that the patients under rhIFN-alpha2b therapy produced neutralized antibody against the IFN. The biochip format would offer a competitive alternative tool not only for facilitating characterization of IFN subtypes but also potentially for enabling clinical serum detection of corresponding antibodies directed against IFNs. PMID:20230300

  9. Screening and Monitoring Coeliac Disease: Multicentre Trial of a New Serum Antibody Test Kit

    Directory of Open Access Journals (Sweden)

    Peter L. Devine

    1994-01-01

    average interassay CV was 6.4% for IgA and 4.3% for IgG (n=3. By defining a positive te st as both IgA and IgG elevated, a sensitivity of 93% in untreated coeliacs (n=75 was observed. The corresponding specificities in healthy adults (n=130 and healthy children (n=77 were >99% and 100% respectively, while in patients with other gastrointestinal disorders (disease controls the specificity was 94% (n=129. The test was also useful in monitoring patients, with anti-gliadin IgA and IgG falling for up to a year after commencing a gluten-free diet (GFD (12 adults. In some patients however, antibody levels did not reach the normal cutpoint after many months on a GFD, which may reflect the patients ' poor adherence to their gluten free diet. The test was superior to the Pharmacia anti-gliadin ELISA, and should be useful as an aid to the diagnosis of coeliac disease, as well as in the follow-up of treated patients.

  10. Engineered Bovine Antibodies in the Development of Novel Therapeutics, Immunomodulators and Vaccines

    Directory of Open Access Journals (Sweden)

    Madhuri Koti

    2014-05-01

    Full Text Available Some bovine antibodies across all classes are unique, such as the CDR3 of the variable heavy-domain (VH CDR3, which is exceptionally long (up to 66 amino acids, unlike most conventional antibodies where the VH CDR3 loops range from 10 to 25 amino acids. The exceptionally long VH CDR3 is encoded by unusually long germline IGHD genes together with insertion of novel “a” nucleotide rich conserved short nucleotide sequence (CSNS specifically at the IGH V-D junction. Such an exceptionally long VH CDR3 confers unique “knob and stalk” structural architecture where the knob, formed by intra-VH CDR3 disulfide bridges, is separated by 20 Å solvent exposed stalk composed of anti-parallel beta strands. The substitution of the knob with cytokines, such as, erythropoietin and granulocyte colony stimulating factor 3 (granulocyte colony stimulating factor, results in expression of functional fusion proteins with enhanced pharmacokinetics. The beta stranded stalk can be substituted with other rigid structures, for example, repeat alpha helices to form coiled-coil that mimics the beta-stranded stalk and, thus, opens opportunities for insertion of this structure in the CDRs of antibodies across species. Given the versatility of such a structural platform in bovine antibody VH CDR3, it provides the opportunity for the development of new generation of diagnostics, therapeutics, vaccines and immunomodulating drugs.

  11. Development of antibody-based c-Met inhibitors for targeted cancer therapy

    Directory of Open Access Journals (Sweden)

    Lee D

    2015-02-01

    Full Text Available Dongheon Lee, Eun-Sil Sung, Jin-Hyung Ahn, Sungwon An, Jiwon Huh, Weon-Kyoo You Hanwha Chemical R&D Center, Biologics Business Unit, Daejeon, Republic of Korea Abstract: Signaling pathways mediated by receptor tyrosine kinases (RTKs and their ligands play important roles in the development and progression of human cancers, which makes RTK-mediated signaling pathways promising therapeutic targets in the treatment of cancer. Compared with small-molecule compounds, antibody-based therapeutics can more specifically recognize and bind to ligands and RTKs. Several antibody inhibitors of RTK-mediated signaling pathways, such as human epidermal growth factor receptor 2, vascular endothelial growth factor, epidermal growth factor receptor or vascular endothelial growth factor receptor 2, have been developed and are widely used to treat cancer patients. However, since the therapeutic options are still limited in terms of therapeutic efficacy and types of cancers that can be treated, efforts are being made to identify and evaluate novel RTK-mediated signaling pathways as targets for more efficacious cancer treatment. The hepatocyte growth factor/c-Met signaling pathway has come into the spotlight as a promising target for development of potent cancer therapeutic agents. Multiple antibody-based therapeutics targeting hepatocyte growth factor or c-Met are currently in preclinical or clinical development. This review focuses on the development of inhibitors of the hepatocyte growth factor/c-Met signaling pathway for cancer treatment, including critical issues in clinical development and future perspectives for antibody-based therapeutics. Keywords: hepatocyte growth factor, ligands, receptor tyrosine kinase, signaling pathway, therapeutic agent

  12. Clinical factors associated with a Candida albicans Germ Tube Antibody positive test in Intensive Care Unit patients

    Directory of Open Access Journals (Sweden)

    Martín-Mazuelos Estrella

    2011-03-01

    Full Text Available Abstract Background Poor outcomes of invasive candidiasis (IC are associated with the difficulty in establishing the microbiological diagnosis at an early stage. New scores and laboratory tests have been developed in order to make an early therapeutic intervention in an attempt to reduce the high mortality associated with invasive fungal infections. Candida albicans IFA IgG has been recently commercialized for germ tube antibody detection (CAGTA. This test provides a rapid and simple diagnosis of IC (84.4% sensitivity and 94.7% specificity. The aim of this study is to identify the patients who could be benefited by the use of CAGTA test in critical care setting. Methods A prospective, cohort, observational multicentre study was carried out in six medical/surgical Intensive care units (ICU of tertiary-care Spanish hospitals. Candida albicans Germ Tube Antibody test was performed twice a week if predetermined risk factors were present, and serologically demonstrated candidiasis was considered if the testing serum dilution was ≥ 1:160 in at least one sample and no other microbiological evidence of invasive candidiasis was found. Results Fifty-three critically ill non-neutropenic patients (37.7% post surgery were included. Twenty-two patients (41.5% had CAGTA-positive results, none of them with positive blood culture for Candida. Neither corrected colonization index nor antifungal treatment had influence on CAGTA results. This finding could corroborate that the CAGTA may be an important biomarker to distinguish between colonization and infection in these patients. The presence of acute renal failure at the beginning of the study was more frequent in CAGTA-negative patients. Previous surgery was statistically more frequent in CAGTA-positive patients. Conclusions This study identified previous surgery as the principal clinical factor associated with CAGTA-positive results and emphasises the utility of this promising technique, which was not

  13. Antibody responses measured by various serologic tests in pigs orally inoculated with low numbers of Toxoplasma gondii oocysts

    DEFF Research Database (Denmark)

    Dubey, J. P.; Andrews, C.D.; Lind, Peter; Kwok, O.C.H.; Thulliez, P.; Lunney, J.K.

    1996-01-01

    Objective-To follow antibody responses measured by various serologic tests in pigs orally inoculated with low (less than or equal to 10 oocysts) numbers of Toxoplasma gondii oocysts. Animals-24, 2- to 3-month-old pigs. Procedure-Pigs (n = 42) were inoculated orally with 10 (14 pigs) or 1 (28 pigs...

  14. Development of an ELISA kit using monoclonal antibody to Clostridium difficile toxin A

    Institute of Scientific and Technical Information of China (English)

    Si-Wu Fu; Ya-Li Zhang; Dian-Yuan Zhou

    2004-01-01

    AIM: To establish an ELISA kit using monoclonal antibodiesagainst Clostridium difficile ( C. difficile) toxin A.METHODS: An indirect sandwich ElISA was described using the purified rabbit monospecific antiserum as capturing antibody. After the polystyrene microtitre plates with 96 fiat-bottomed wells were coated with rabbit antiserum, the wells were blocked with 100 g/L BSA in PBS-T. C. difficiletoxin A or culture filtrates were added to each well and then monoclonal antibodies IgG-horseradish peroxidase conjugate was added as detecting antibody, tetramethylbenzidine was used as substrate and A450 of the stopped reacting product was recorded in an automated plate reader. RESULTS: The tested specimens included culture filtrates of 2 strains of toxigenic C. difficile, 2 strains of non-toxigenic C. difficile, 26 strains of E. coli, 2 strains of S. dysenteriae, 1 strain of Bif infantis, 5 strains of V. cholera, 2 strains ofS. typhi, 7 strains of C. botulinum, 1 strain of toxigenic C. sordllii, and 1 strain of C. butyricum. A total of 47 strains of culture filtrates were all negative except for 2 strains of toxigenic C. difficile. The detective limitation of toxin A was 0.1 ng/mL.CONCLUSION: An ELISA kit with high specificity and excellent sensitivity for the rapid detection of C. difficile toxin A was established. It will be a useful tool for diagnostic test of C. difficile toxin A.

  15. Naturally acquired Lawsonia intracellularis infection in pigs studied from weaning to slaughter by indirect immunofluorescence antibody test and polymerase chain reaction on faeces

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre; Vigre, Håkan; Sørensen, Vibeke; Møller, Kristian

    2005-01-01

    pigs were 2-4 weeks old maternally derived IgG against L. intracellularis was demonstrated by immunofluorescence antibody test in nine pigs whereas the bacterium was detected by PCR in faeces from six pigs. The maternally derived antibodies did not prevent pigs from becoming infected as seven pigs...... immunofluorescence antibody test compared to 24% by immunohistochemistry on ileal samples. Thus, applied at the time of slaughter the antibody test appeared to be a highly sensitive ante-mortem diagnostic tool for identifying L. intracelluaris exposed pigs with or without current proliferative enteropathy. (c) 2004...

  16. Development of a nanogold-based immunochromatographic assay for detection of morphine in urine using the Amor-HK16 monoclonal antibody.

    Science.gov (United States)

    Dehghannezhad, Ardeshir; Paknejad, Maliheh; Rasaee, Mohammad Javad; Omidfar, Kobra; Seyyed Ebrahimi, Shadi Sadat; Ghahremani, Hossein

    2012-12-01

    A simple, rapid competitive immunochromatography (ICG) strip test was developed to detect morphine in urine samples using a monoclonal antibody produced in-house and conjugated to gold nanoparticles. Hybridoma cells were cultured and the Amor-HK16 monoclonal antibody against morphine was obtained from the supernatant after purification by salting out and passing through a Protein G-Agarose affinity column. Morphine was obtained from morphine sulfate and a C6-hemisuccinate derivative of morphine was prepared, conjugated to bovine serum albumin, and immobilized to a nitrocellulose membrane as the test line. Goat anti-mouse antibody was used as a binder in the control line in the detection zone of the strip. Colloidal gold particles of diameter approximately 20 nm were prepared and conjugated to the monoclonal antibody. The detection limit of the test strip was found to be 2000 ng/mL of morphine in urine samples. Reliability was determined by performing the ICG test on 103 urine samples and comparing the results with those obtained by thin-layer chromatography. The sensitivity of the test was 100%, and the analysis time for the assay was approximately 5 min. The new ICG method was adequately sensitive and accurate for the rapid screening of morphine in urine. PMID:23244319

  17. Rock support system development test plan

    International Nuclear Information System (INIS)

    The Test Plan has been prepared to support design activities for the development of a rock support system for a Nuclear Waste Repository in Basalt (NWRB). The rock support system is assumed to consist of a combination of shotcrete and rock bolts. The seven testing activities include mix development and physical testing of shotcrete, durability testing of shotcrete, durability testing of rock bolt grouts, field tests on rock bolts, field testing of shotcrete, and heated room test. The objective of the Test Plan is to develop required data through combined laboratory, field, and office studies for design and design validation of the rock support system. The overall Test Plan is developed to provide a logical progression from laboratory tests performed to characterize fundamental thermomechanical properties of shotcrete and grouts, to field tests on rock bolts and shotcrete, and in situ performance tests. 21 refs., 15 figs., 33 tabs

  18. Development, characterization and diagnostic application of a monoclonal antibody specific for a proteinase K resistant Lawsonia intracellularis antigen

    DEFF Research Database (Denmark)

    Boesen, Henriette T.; Jensen, Tim Kåre; Jungersen, Gregers;

    2005-01-01

    purposes (by immunohistochemistry) and for bacterial characterization. Several antibody producing hybridomas were established by fusion of mouse myeloma with spleen cells from BALB/c mice immunized with mucosa scrapings of the intestinal mucosa from a L. intracellularis infected pig. A monoclonal antibody...... (mAb), Law1-DK, isotyped as IgG2b was selected by indirect immunofluorescence antibody test (IFAT). Histological sections of the intestines from pigs affected by proliferative enteropathy and in vitro grown bacteria in cell culture were tested positive for the presence of L. intracellularis with the...

  19. Seroprevalence of Plasmodium vivax in the Republic of Korea (2003-2005) using Indirect Fluorescent Antibody Test

    OpenAIRE

    Kim, Tong-Soo; Kang, Yoon-Joong; Lee, Won-Ja; Na, Byoung-Kuk; Moon, Sung-Ung; Cha, Seok Ho; Lee, Sung-Keun; Park, Yun-Kyu; Pak, Jhang-Ho; Cho, Pyo Yun; Sohn, Youngjoo; Lee, Hyeong-Woo

    2014-01-01

    Plasmodium vivax reemerged in the Republic of Korea (ROK) in 1993, and is likely to continue to affect public health. The purpose of this study was to measure levels of anti-P. vivax antibodies using indirect fluorescent antibody test (IFAT) in border areas of ROK, to determine the seroprevalence of malaria (2003-2005) and to plan effective control strategies. Blood samples of the inhabitants in Gimpo-si, Paju-si, and Yeoncheon-gun (Gyeonggi-do), and Cheorwon-gun (Gangwon-do) were collected a...

  20. An experimental test of stroke recovery by implanting a hyaluronic acid hydrogel carrying a Nogo receptor antibody in a rat model

    Energy Technology Data Exchange (ETDEWEB)

    Ma Jun [Biomaterials Laboratory, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Tian Weiming [Biomaterials Laboratory, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Hou Shaoping [Beijing Institute of Neuroscience, Capital University of Medical Sciences, Beijing 100054 (China); Xu Qunyuan [Beijing Institute of Neuroscience, Capital University of Medical Sciences, Beijing 100054 (China); Spector, Myron [Tissue Engineering, VA Boston Healthcare System, Harvard Medical School, Boston, MA (United States); Cui Fuzhai [Biomaterials Laboratory, Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China)

    2007-12-15

    The objective of the study was to determine the effects of a hyaluronic-acid-based (HA-based) hydrogel implant, carrying a polyclonal antibody to the Nogo-66 receptor (NgR), on adult rats that underwent middle cerebral artery occlusion (MCAO). Behavioral tests of a forelimb-reaching task suggested that the disabled function of the impaired forelimb in this stroke model was ameliorated by the implant to a certain extent. These behavioral findings were correlated with immunohistochemical results of investigating the distribution of NgR antibody, neurofilaments (NF) and neuron-specific class III {beta}-tubulin (TuJ1) in the brain sections. The porous hydrogel functioned as a scaffold to deliver the NgR antibody, support cell migration and development. In addition, it was found NF-positive and TuJ1-positive expressions were distributed in the implanted hydrogel. Collectively, the results demonstrate the promise of the HA hydrogel as a scaffold material and the delivery vehicle of the NgR antibody for the repair of defects and the support of neural regeneration in the brain.

  1. An experimental test of stroke recovery by implanting a hyaluronic acid hydrogel carrying a Nogo receptor antibody in a rat model

    International Nuclear Information System (INIS)

    The objective of the study was to determine the effects of a hyaluronic-acid-based (HA-based) hydrogel implant, carrying a polyclonal antibody to the Nogo-66 receptor (NgR), on adult rats that underwent middle cerebral artery occlusion (MCAO). Behavioral tests of a forelimb-reaching task suggested that the disabled function of the impaired forelimb in this stroke model was ameliorated by the implant to a certain extent. These behavioral findings were correlated with immunohistochemical results of investigating the distribution of NgR antibody, neurofilaments (NF) and neuron-specific class III β-tubulin (TuJ1) in the brain sections. The porous hydrogel functioned as a scaffold to deliver the NgR antibody, support cell migration and development. In addition, it was found NF-positive and TuJ1-positive expressions were distributed in the implanted hydrogel. Collectively, the results demonstrate the promise of the HA hydrogel as a scaffold material and the delivery vehicle of the NgR antibody for the repair of defects and the support of neural regeneration in the brain

  2. High Affinity, Developability and Functional Size: The Holy Grail of Combinatorial Antibody Library Generation

    Directory of Open Access Journals (Sweden)

    Kathrin Tissot

    2011-05-01

    Full Text Available Since the initial description of phage display technology for the generation of human antibodies, a variety of selection methods has been developed. The most critical parameter for all in vitro-based approaches is the quality of the antibody library. Concurrent evolution of the libraries has allowed display and selection technologies to reveal their full potential. They come in different flavors, from naïve to fully synthetic and differ in terms of size, quality, method of preparation, framework and CDR composition. Early on, the focus has mainly been on affinities and thus on library size and diversity. Subsequently, the increased awareness of developability and cost of goods as important success factors has spurred efforts to generate libraries with improved biophysical properties and favorable production characteristics. More recently a major focus on reduction of unwanted side effects through reduced immunogenicity and improved overall biophysical behavior has led to a re-evaluation of library design.

  3. DEVELOPMENT OF ANTIBODY TO RALSTONIA SOLANACEARUM AND ITS APPLICATION FOR DETECTION OF BACTERIAL WILT

    Directory of Open Access Journals (Sweden)

    YADI SURYADI

    2009-01-01

    Full Text Available The serological assay for the detection of bacterial wilt caused by Ralstonia solanacearum (RSwas able to provide information regarding the presence of the pathogen in plant materials. Theresearch is was aimed to develop polyclonal antibody (PAb for RS detection. Bacterial wholecells of RS isolates mixed with glutaraldehyde were used to immunize New Zealand femalewhite rabbit. The titre of antibody in culture supernatant was 1: 1024. The PAb developed froma ground nut RS isolates reacted with infected plant samples from various locations. It was ableto detect RS antigen of crude extract and pure cultures from tomato and potato plant samples4-5using dot blot ELISA; however, the minimum detectable concentration of RS antigen was 10cells/ml. The PAb obtained in this study is sensitive enough to detect RS isolates in routineserological assay

  4. Development of a novel monoclonal antibody to B7-H4: characterization and biological activity

    Directory of Open Access Journals (Sweden)

    Qian Y

    2011-07-01

    Full Text Available Abstract Objective B7-H4, a member of the B7 family of immunoregulatory receptors, may participate in the negative regulation of cell-mediated immunity. Aberrant B7-H4 expression is detected in some tumors and it plays a role in the occurrence and development of tumors. The aim of this study was to elucidate the functional and structural properties of B7-H4. Methods We developed a monoclonal antibody (mAb against the extracellular domain of B7-H4 through immunization of Balb/c mice with 3T3-mB7-H4 cells which expressed extrinsic B7-H4. A stable hybridoma cell line was established. Then, we analysised the characterization of the mAb through Enzyme linked immunosorbent assay (ELISA, Immunoprecipitation (IP, western blotting, Immunohistochemical (IHC, and tested the biological activity of the mAb. Results ELISA, IP, and western blotting analyses indicated that the mAb specifically recognized B7-H4. In addition, flow cytometry demonstrated that the mAb exhibits excellent reactivity when applied to leukemic cells. IHC staining revealed that the mAb stained in a predominantly diffuse plasmalemmal or cytoplasmic pattern when applied to certain tumor tissues. The preliminary results of the mAb's biological activity showed that the mAb could effectively inhibit the function of B7-H4 in the inhibition of T cell, while promotingg the growth of T cells and the secretion of Interleukin-2 (lL-2, Interleukin-4 (IL-4, Interleukin10 (IL-10 and Interferon-γ (IFN-γ. Conclusion This mAb will be a valuable tool for the further investigation of B7-H4 function.

  5. How (Much) Do Developers Test?

    NARCIS (Netherlands)

    Beller, M.; Gousios, G.; Zaidman, A.E.

    2015-01-01

    What do we know about software testing in the real world? It seems we know from Fred Brooks’ seminal work “The Mythical Man-Month” that 50% of project effort is spent on testing. However, due to the enormous advances in software engineering in the past 40 years, the question stands: Is this observat

  6. Vocabulary Development: Teaching vs. Testing.

    Science.gov (United States)

    Nilsen, Alleen Pace; Nilsen, Don L. F.

    2003-01-01

    Considers how with no training in how to teach vocabulary skills, many teachers transfer to their classroom the same techniques that they see test makers using. Offers a chart to encourage thinking about the ways that standardized testing techniques differ from good teaching and learning practices. Argues that educators should provide students…

  7. Novel Point-of-Care Test for Simultaneous Detection of Nontreponemal and Treponemal Antibodies in Patients with Syphilis ▿

    OpenAIRE

    Castro, Arnold R.; Esfandiari, Javan; Kumar, Shailendra; Ashton, Matthew (British painter, active from 1728); Kikkert, Susan E.; Park, Mahin M.; Ballard, Ronald C.

    2010-01-01

    We describe a point-of-care immunochromatographic test for the simultaneous detection of both nontreponemal and treponemal antibodies in the sera of patients with syphilis that acts as both a screening and a confirmatory test. A total of 1,601 banked serum samples were examined by the dual test, and the results were compared to those obtained using a quantitative rapid plasma reagin (RPR) test and the Treponema pallidum passive particle agglutination (TP-PA) assay. Compared to the RPR test, t...

  8. Brucella Antibody Titer (Wright’s Test in Healthy Primary School Children in Tehran

    Directory of Open Access Journals (Sweden)

    A Zamani

    2005-10-01

    Full Text Available Background: Brucellosis is a common worldwide disease of both humans and animals. Iran is one of the endemic areas infected with Brucellosis and a prevalence of 225 in 100,000. Early diagnosis and prompt treatment will save the child from death and disabilities. Considering that Wrights test is easy, feasible and fast to use, it is one of the most common tests used in the diagnosis of Brucellosis. It has a high sensitivity but a low specificity. A titer >1/160 is considered positive. Due to the prevalence of Brucellosis in Iran a titer >1/80 in suspected patients should be a matter of concern and further investigation. Methods: This prospective cross-sectional descriptive analytical study was performed with the aim to determine Brucella antibody level (Wright's test in healthy primary school children in 19 educational sectors of Tehran. Findings: A total of 1531 children were enrolled, 674(44% and 857 (56% of whom were boys and girls with a mean age of 9.171.55 years. 99.7% of the children had a titer less than 1/80 and 5 children (0.3% had a titer ≥1/160. There was a significant statistical relation between Wright's titer and gender (p=0.005. There was, however, no significant statistical relation to age (p=0.4 or different geographical regions of Tehran (p=0.2. Conclusion: The rate of Brucella infection in school children in Tehran was unremarkable. According to the results of our study, particularly in endemic areas, a Wright's titer of 1:80 in suspected cases for Brucellosis can be taken as a diagnostic titer.

  9. Stabilization and development of sustained-release formulations of protein/antibody for subcutaneous delivery

    OpenAIRE

    Marquette, Sarah

    2014-01-01

    ABSTRACTThis project aimed at developing a drug delivery system (DDS) able to enhance the stability andresidence time in vivo of antibodies (Abs). The system will deliver drug by the subcutaneousroute (SC), while ensuring accurate control of the drug release and the resulting plasmatic level. This technology platform will allow to reduce frequency of injection, potentially decrease side effects and maintain high concentration of Abs which will improve life of patient having chronic disease su...

  10. Development and characterization of monoclonal antibodies to Marek's disease tumor-associated surface antigen.

    OpenAIRE

    Liu, X. F.; Lee, L F

    1983-01-01

    Four monoclonal antibodies, A35, B94, EB29, and G152, against Marek's disease tumor-associated surface antigen have been developed and their specificities studied against a panel of Marek's disease and lymphoid leukosis primary tumors; Marek's disease, and lymphoid leukosis, and reticuloendotheliosis lymphoblastoid cell lines; and normal chicken cells. A35 and G152 are of the immunoglobulin M class, and B94 and EB29 are of the immunoglobulin G1 subclass.

  11. Development of a monoclonal antibody detection assay for species-specific identification of abalone.

    Science.gov (United States)

    Lopata, Andreas L; Luijx, Thomas; Fenemore, Bartha; Sweijd, Neville A; Cook, Peter A

    2002-10-01

    Species identification based on biochemical and molecular techniques has a broad range of applications. These include compliance enforcement, the management and conservation of marine organisms, and commercial quality control. Abalone poaching worldwide and illegal trade in abalone products have increased mainly because of the attractive prices obtained and caused a sharp decline in stocks. Alleged poachers have been acquitted because of lack of evidence to correctly identify species. Therefore, a robust method is required that would identify tissue of abalone origin to species level. The aim of this study was to develop immunologic techniques, using monoclonal and polyclonal antibodies, to identify 10 different abalone species and subspecies from South Africa, the United States, Australia, and Japan. The combination of 3 developed monoclonal antibodies to South African abalone (Haliotis midae) enabled differentiation between most of the 10 species including the subspecies H. diversicolor supertexta and H. diversicolor diversicolor. In a novel approach, using antibodies of patients with allergy to abalone, the differentiation of additional subspecies, H. discus discus and H. discus hannai, was possible. A field-based immunoassay was developed to identify confiscated tissue of abalone origin. PMID:14961238

  12. Development of a double-antibody sandwich ELISA for rapid detection of Bacillus Cereus in food.

    Science.gov (United States)

    Zhu, Longjiao; He, Jing; Cao, Xiaohan; Huang, Kunlun; Luo, Yunbo; Xu, Wentao

    2016-01-01

    Bacillus cereus is increasingly recognized as one of the major causes of food poisoning in the industrialized world. In this paper, we describe a sensitive double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) that was developed for rapid detection of B. cereus in food to minimize the risk of contamination. The polyclonal antibody (pAb) and monoclonal antibodies (mAbs) specific to B. cereus were generated from rabbit antiserum and mouse ascites, respectively, using the octanoic acid/saturated ammonium sulfate precipitation method and protein A-sepharose columns. IgG-isotype mAbs were specially developed to undergo a novel peripheral multiple sites immunization for rapid gain of hybridomas and a subtractive screen was used to eliminate cross reactivity with closely related species such as Bacillus thuringiensis, B. subtilis, B. licheniformis and B. perfringens. The linear detection range of the method was approximately 1 × 10(4)-2.8 × 10(6) cells/mL with a detection limit (LOD) of 0.9 × 10(3) cells/mL. The assay was able to detect B. cereus when the samples were prepared in meat with various pathogens. The newly developed analytical method provides a rapid method to sensitively detect B. cereus in food specimens. PMID:26976753

  13. Genital chlamydial infection among women in Nicaragua: validity of direct fluorescent antibody testing, prevalence, risk factors and clinical manifestations.

    OpenAIRE

    B. Herrmann; Espinoza, F.; Villegas, R R; Smith, G.D.; Ramos, A.; Egger, M.

    1996-01-01

    OBJECTIVE: To validate the performance of a direct fluorescence antibody (DFA) test and to determine the prevalence, risk factors and clinical manifestations of cervical chlamydia infection in different groups of women in Nicaragua. STUDY POPULATION: 926 women, 863 routine clinic attenders (mean age 27 years) and 63 sex workers (mean age 25 years) attending health centres in León, Corinto, Matagalpa and Bluefields. METHODS: Cervical specimens were examined using the Syva MicroTrak test system...

  14. Antibodies to the enterobacterial common antigen: standardization of the passive hemagglutination test and levels in normal human sera.

    OpenAIRE

    Malkamäki, M

    1981-01-01

    The passive hemagglutination test for antibodies against the enterobacterial common antigen (ECA) of Kunin was standardized for diagnostic purposes. Human erythrocytes were coated with a soluble ECA+ preparation from Salmonella typhimurium or, as specificity controls, with a similar ECA- preparation from congenic ECA-negative bacteria with saline, and the hemagglutination assay was performed on microtiter plates. The specificity of the test was ascertained further by inhibition assays with pu...

  15. Studies using tests to determine antibodies against non-structural proteins of foot and mouth disease in Colombia

    International Nuclear Information System (INIS)

    Colombia is a country with free areas without vaccination; free areas with vaccination and endemic areas. Buffer regions also are well surveyed. The various geographical areas where the present vaccination and control policies are being made are being revised in the light of animal movement control and surveillance. The role of NSP antibody testing (fitness for purpose) is very pertinent. The laboratory involved in FMD control is capable of antigen detection through tissue culture antigen capture ELISA; CFT; PCR; probang testing; (carriers) and antibody testing through VNT, VIAA AGID; LPBE and various NSP tests including the PANAFTOSA system using a 3ABC ELISAs and IETB. So we have a complete system for any studies in cattle and pigs. The routine work includes surveillance of cattle both from zones with vaccination and without disease and also testing animals that are moved from buffer zones into vaccination but disease free areas. In 2003, 13,016 and 10,000 samples respectively for the two situations were examined. Results of testing from different populations and comparison of tests are made. Various numbers of sera were tested with a variety of tests including: United Biomedical Incorporated (UBI) NSP ELISA test Chekit-FMD-3ABC (then BOMMELI diagnostics) CEDI test FMDV-NSP (CEDI) I-ELISA-3ABC from PANAFTOSA (IELISA) Electroimmunetransfer blot (EITB) VIAA Immunodiffusion test (VIAA AGID). (author)

  16. Test-driven development with Django

    CERN Document Server

    Harvey, Kevin

    2015-01-01

    This book is for Django developers with little or no knowledge of test-driven development or testing in general. Familiarity with the command line, setting up a Python virtual environment, and starting a Django project are assumed.

  17. Development and validation of the 57Co assay for determining the ligand to antibody ratio in bifunctional chelate/antibody conjugates for use in radioimmunotherapy

    International Nuclear Information System (INIS)

    Introduction: The ligand to antibody ratio is an important characteristic of a chelate/antibody conjugate. It has been widely reported that if the ratio is too high, there will be detrimental effects on immunoreactivity and biodistribution; conversely, if the ratio is too low, the radionuclide may not bind efficiently, and the stability and the specific activity will be reduced. There are little published data on the accuracy or precision of the 57Co assay. The UK Clinical Trials Regulations state that “systems with procedures that assure the quality of every aspect of the trial should be implemented”. The aims of this study were to assess the reliability and accuracy of the 57Co binding assay and validate it against defined criteria. Method: Thirty-two serial assays were assessed for reliability. Two batches of conjugated antibody were also analysed by matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) mass spectrometry (MS) to allow the comparison of the functional test with a physical method. Results: Reliability: The coefficient of variation was 0.13. Accuracy: There was 9% variation between the 57Co binding assay and MALDI-TOF MS results. Conclusion: A detailed method for the 57Co ligand to antibody test is described that allows a discrete value to be obtained. The assay was validated as fit for purpose against target values of coefficient of variation <0.20, accuracy±10%, over a permissive range of 0.5–3.0 ligand to antibody ratio.

  18. Evaluation of Anti-Nuclear antibody test results in clinical practice

    Directory of Open Access Journals (Sweden)

    Nevreste Çelikbilek

    2015-06-01

    Full Text Available Objective: Aim of this study is to evaluate anti-nuclear antibody (ANA test results obtained between 2009 and 2011. Methods: Of a totally 5068 cases tested for ANA by indirect immunofluorescence method (IIFA, randomly chosen 982 ANA-positive cases were reviewed in terms of gender, level and pattern of fluorescence, anti-dsDNA (anti-double stranded DNA and anti-extractable nuclear antigen (ENA profile. Anti-dsDNA levels and anti-ENA profiles were determined by enzyme linked immune assay (ELISA and immune-blotting (IB, respectively. Results: Sex distribution of ANA positive patients was determined as 756 (77% females and 226 (23% males. Fifty per cent of the cases were from rheumatology department, 20% from gastroenterology and 30% from other units. Fluorescence levels were considered borderline or weak positive in 62.6% of the samples. The most frequent patterns were homogeneous (23%, speckled (22%, homogeneous-speckled (15.5% and nucleolar (13.5%. Anti-dsDNA were studied in 759 ANA positive patients and 66 (8.7% samples were found positive, being 44 of them (68.8% with homogeneous pattern and the rest with speckled, nucleolar, nuclear dots, centromeric or midbody patterns. Totally 131 (31.6% of 414 samples studied for anti-ENA profile were found positive. The first four frequent profiles were SSA (34.4%, SSA-SSB (16.8%, Scl70 (16% and Sm/RNP (9.2%. Conclusion: Our results are similar with the current related literature. It is known that autoantibodies can be detectable before clinical symptoms being apparent, especially in SLE. Therefore, borderline or weak fluorescence levels should also be reported and the patients having them should be followed-up carefully. J Microbiol Infect Dis 2015;5(2: 63-68

  19. Development of a mixed antigen agar gel enzyme assay (AGEA) for the detection of antibodies to poxvirus in chicken and turkey sera.

    Science.gov (United States)

    Tadese, Theodros; Potter, E A; Reed, W M

    2003-02-01

    A mixed-antigen agar gel enzyme assay (AGEA) was developed to detect antibodies to poxviruses in chicken and turkey sera. The assay combines the principles of immunodiffusion and enzyme assay. For the detection of antibodies to fowl poxvirus (FP), pigeon poxvirus (PP) and turkey poxvirus (TP) in turkey serum samples, the three antigens were combined to form a mixed-antigen assay. To screen for antibodies to FP and PP in chicken serum samples, the two antigens were combined. When FP and PP viruses were combined as antigens, the sensitivity for chicken sera was 64% but the sensitivity of the agar gel precipitation test (AGPT) was 34% (PAGEA had a sensitivity of 66.4% while that of AGPT was 25% (P<0.001). PMID:12655123

  20. Mobile Router Developed and Tested

    Science.gov (United States)

    Ivancic, William D.

    2002-01-01

    The NASA Glenn Research Center, under a NASA Space Act Agreement with Cisco Systems, has been performing joint networking research to apply Internet-based technologies and protocols to space-based communications. As a result of this research, NASA performed stringent performance testing of the mobile router, including the interaction of routing and the transport-level protocol. In addition, Cisco Systems developed the mobile router for both commercial and Government markets. The code has become part of the Cisco Systems Internetworking Operating System (IOS) as of release 12.2 (4) T--which will make this capability available to the community at large. The mobile router is software code that resides in a network router and enables entire networks to roam while maintaining connectivity to the Internet. This router code is pertinent to a myriad of applications for both Government and commercial sectors, including the "wireless battlefield." NASA and the Department of Defense will utilize this technology for near-planetary observation and sensing spacecraft. It is also a key enabling technology for aviation-based information applications. Mobile routing will make it possible for information such as weather, air traffic control, voice, and video to be transmitted to aircraft using Internet-based protocols. This technology shows great promise in reducing congested airways and mitigating aviation disasters due to bad weather. The mobile router can also be incorporated into emergency vehicles (such as ambulances and life-flight aircraft) to provide real-time connectivity back to the hospital and health-care experts, enabling the timely application of emergency care. Commercial applications include entertainment services, Internet protocol (IP) telephone, and Internet connectivity for cruise ships, commercial shipping, tour buses, aircraft, and eventually cars. A mobile router, which is based on mobile IP, allows hosts (mobile nodes) to seamlessly "roam" among various IP

  1. Effect of haemolysis and repeated freeze-thawing cycles on wild boar serum antibody testing by ELISA

    Directory of Open Access Journals (Sweden)

    Boadella Mariana

    2011-11-01

    Full Text Available Abstract Background Monitoring wildlife diseases is needed to identify changes in disease occurrence. Wildlife blood samples are valuable for this purpose but are often gathered haemolysed. To maximise information, sera often go through repeated analysis and freeze-thaw cycles. Herein, we used samples of clean and haemolysed Eurasian wild boar (Sus scrofa serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV. Results The estimated prevalence of antibodies against ADV was 50-53% for clean and haemolysed sera. Hence, haemolysis did not reduce the mean observed serum antibody prevalence. However, 10 samples changed their classification after repeated freeze-thawing. This included 3 (15% of the clean sera and 7 (41% of the haemolysed sera. Conclusions We recommend (1 establishing more restrictive cut-off values when testing wildlife sera, (2 recording serum quality prior to sample banking, (3 recording the number of freezing-thawing cycles and (4 store sera in various aliquots to reduce repeated usage. For instance, sera with more than 3 freeze-thaw cycles and a haemolysis of over 3 on a scale of 4 should better be discarded for serum antibody monitoring. Even clean (almost not haemolysed sera should not go through more than 5 freeze-thaw cycles.

  2. TB-SA antibody test for diagnosis and monitoring treatment outcome of sputum smear negative pulmonary tuberculosis patients.

    Science.gov (United States)

    Li, Xinxu; Xu, Hancheng; Jiang, Shiwen; Jing, Kuanhe; Wang, Li; Liu, Xiaoqiu; Li, Weibin; Zhang, Hui; Wang, Lixia

    2011-09-01

    The objectives of this study were to evaluate the suitability of the TB-SA antibody test to diagnose tuberculosis in sputum smear negative (SS-) pulmonary tuberculosis (TB) patients and its applicability for monitoring treatment outcomes in these patients. This study was conducted in three counties/districts in Chongqing Municipality, Liaoning Province, China between June 2005 and June 2007. A total of 432 SS suspected pulmonary TB patients were recruited and their blood was collected prior to treatment, at the end of 1 month of treatment, 2 months of treatment and 6 months of treatment (E6MT). The serum samples were analyzed with a TB-SA antibody test kit. Of the 432 SS suspected pulmonary TB patients, serum samples were obtained at all time points in 316 patients and analyzed. The 316 patients were divided into three groups according to sputum smear and sputum culture results and the chest X-ray results before treatment and at E6MT. Ten point four percent were SS-/culture positive (C+), 73.1% were SS-/culture negative (C-) with X-rays abnormalities, and 16.5% were SS-/C- without X-rays abnormalities. The positive rates for TB-SA antibody in the three groups were 57.6, 44.6 and 44.2%, respectively, before treatment, and 18.2, 19.1 and 26.9%, respectively, at E6MT. There was a significant decrease in TB-SA antibody positivity with treatment for all 3 groups. The TB-SA antibody test may be a useful adjunct to diagnose tuberculosis in SS- pulmonary TB patients, and may be useful for monitoring treatment outcomes of SS- pulmonary TB patients. PMID:22299440

  3. Development of monoclonal antibodies specifically recognizing the cyst stage of Entamoeba histolytica.

    Science.gov (United States)

    Walderich, B; Burchard, G D; Knobloch, J; Müller, L

    1998-09-01

    Protozoan cysts were isolated according to a two-step sucrose gradient procedure. Pure cysts of Entamoeba histolytica, in fixed and native states, were injected into BALB/c mice intraperitoneally for immunization. The spleens of these animals were used for fusion with AG8 mouse myeloma cells. Hybridomas were obtained and tested for the recognition of E. histolytica, E. dispar, E. coli, E. hartmanni, Endolimax nana, Jodamoeba biitschlii, and Giardia lamblia. Three monoclonal antibodies were identified that reacted only with cysts and trophozoites of E. histolytica. These can be used for differentiation and identification of E. histolytica in feces. PMID:9749623

  4. Waiving in vivo studies for monoclonal antibody biosimilar development: National and global challenges.

    Science.gov (United States)

    Chapman, Kathryn; Adjei, Akosua; Baldrick, Paul; da Silva, Antonio; De Smet, Karen; DiCicco, Richard; Hong, Seung Suh; Jones, David; Leach, Michael W; McBlane, James; Ragan, Ian; Reddy, Praveen; Stewart, Donald I H; Suitters, Amanda; Sims, Jennifer

    2016-01-01

    Biosimilars are biological medicinal products that contain a version of the active substance of an already authorised original biological medicinal product (the innovator or reference product). The first approved biosimilar medicines were small proteins, and more recently biosimilar versions of innovator monoclonal antibody (mAb) drugs have entered development as patents on these more complex proteins expire. In September 2013, the first biosimilar mAb, infliximab, was authorised in Europe. In March 2015, the first biosimilar (Zarxio™, filgrastim-sndz, Sandoz) was approved by the US Food and Drug Administration; however, to date no mAb biosimilars have been approved in the US. There are currently major differences between how biosimilars are regulated in different parts of the world, leading to substantial variability in the amount of in vivo nonclinical toxicity testing required to support clinical development and marketing of biosimilars. There are approximately 30 national and international guidelines on biosimilar development and this number is growing. The European Union's guidance describes an approach that enables biosimilars to enter clinical trials based on robust in vitro data alone; in contrast, the World Health Organization's guidance is interpreted globally to mean in vivo toxicity studies are mandatory. We reviewed our own experience working in the global regulatory environment, surveyed current practice, determined drivers for nonclinical in vivo studies with biosimilar mAbs and shared data on practice and study design for 25 marketed and as yet unmarketed biosimilar mAbs that have been in development in the past 5y. These data showed a variety of nonclinical in vivo approaches, and also demonstrated the practical challenges faced in obtaining regulatory approval for clinical trials based on in vitro data alone. The majority of reasons for carrying out nonclinical in vivo studies were not based on scientific rationale, and therefore the authors

  5. Development of QCM Biosensor with Specific Cow Milk Protein Antibody for Candidate Milk Adulteration Detection

    OpenAIRE

    Sakti, Setyawan P.; Nur Chabibah; Ayu, Senja P.; Masdiana C. Padaga; Aulanni’am Aulanni’am

    2016-01-01

    Adulteration of goat milk is usually done using cow’s milk product. Cow milk is used as it is widely available and its price is cheaper compared to goat milk. This paper shows a development of candidate tools for milk adulteration using cow milk. A quartz crystal microbalance immunosensor was developed using commercial crystal resonator and polyclonal antibody specific to cow milk protein. A specific protein at 208 KDa is found only in cow milk and does not exist in goat milk. The existence o...

  6. Development of Immunoassay Based on Monoclonal Antibody Reacted with the Neonicotinoid Insecticides Clothianidin and Dinotefuran

    Directory of Open Access Journals (Sweden)

    Seiji Iwasa

    2012-11-01

    Full Text Available Enzyme-linked immunosorbent assay (ELISA based on a monoclonal antibody (MoAb was developed for the neonicotinoid insecticide clothianidin. A new clothianidin hapten (3-[5-(3-methyl-2-nitroguanidinomethyl-1,3-thiazol-2-ylthio] propionic acid was synthesized and conjugated to keyhole limpet hemocyanin, and was used for monoclonal antibody preparation. The resulting MoAb CTN-16A3-13 was characterized by a direct competitive ELISA (dc-ELISA. The 50% of inhibition concentration value with clothianidin was 4.4 ng/mL, and the working range was 1.5–15 ng/mL. The antibody showed high cross-reactivity (64% to dinotefuran among the structurally related neonicotinoid insecticides. The recovery examinations of clothianidin for cucumber, tomato and apple showed highly agreement with the spiked concentrations; the recovery rate was between 104% and 124% and the coefficient of variation value was between 1.8% and 15%. Although the recovery rate of the dc-ELISA was slightly higher than that of HPLC analysis, the difference was small enough to accept the dc-ELISA as a useful method for residue analysis of clothianidin in garden crops.

  7. Clinical development of monoclonal antibody-based drugs in HIV and HCV diseases

    Directory of Open Access Journals (Sweden)

    Flego Michela

    2013-01-01

    Full Text Available Abstract Today there are many licensed antiviral drugs, but the emergence of drug resistant strains sometimes invalidates the effects of the current therapies used in the treatment of infectious diseases. Compared to conventional antiviral drugs, monoclonal antibodies (mAbs used as pharmacological molecules have particular physical characteristics and modes of action, and, therefore, they should be considered as a distinct therapeutic class. Despite being historically validated, antibodies may represent a novel tool for combatting infectious diseases. The current high cost of mAbs' production, storage and administration (by injection only and the consequent obstacles to development are outweighed by mAbs' clinical advantages. These are related to a low toxicity combined with high specificity and versatility, which allows a specific antibody to mediate various biological effects, ranging from the virus neutralization mechanisms to the modulation of immune responses. This review briefly summarizes the recent technological advances in the field of immunoglobulin research, and the current status of mAb-based drugs in clinical trials for HIV and HCV diseases. For each clinical trial the available data are reported and the emerging conceptual problems of the employed mAbs are highlighted. This overview helps to give a clear picture of the efficacy and challenges of the mAbs in the field of these two infectious diseases which have such a global impact.

  8. Frequencies and Specificities of “Enzyme-Only” Detected Erythrocyte Alloantibodies in Patients Hospitalized in Austria: Is an Enzyme Test Required for Routine Red Blood Cell Antibody Screening?

    OpenAIRE

    Dietmar Enko; Claudia Habres; Franz Wallner; Barbara Mayr; Gabriele Halwachs-Baumann

    2014-01-01

    The aim of this study was to determine the frequencies and specificities of “enzyme-only” detected red blood cell (RBC) alloantibodies in the routine antibody screening and antibody identification in patients hospitalized in Austria. Routine blood samples of 2420 patients were investigated. The antibody screening was performed with a 3-cell panel in the low-ionic strength saline- (LISS-) indirect antiglobulin test (IAT) and with an enzyme-pretreated (papain) 3-cell panel fully automated on th...

  9. Test-driven development with Mockito

    CERN Document Server

    Acharya, Sujoy

    2013-01-01

    This book is a hands-on guide, full of practical examples to illustrate the concepts of Test Driven Development.If you are a developer who wants to develop software following Test Driven Development using Mockito and leveraging various Mockito features, this book is ideal for you. You don't need prior knowledge of TDD, Mockito, or JUnit.It is ideal for developers, who have some experience in Java application development as well as a basic knowledge of unit testing, but it covers the basic fundamentals of TDD and JUnit testing to get you acquainted with these concepts before delving into them.

  10. Development of two Trichoplusia ni larvae-derived ELISAs for the detection of antibodies against replicase and capsid proteins of porcine circovirus type 2 in domestic pigs.

    Science.gov (United States)

    Pérez-Martín, Eva; Grau-Roma, Llorenç; Argilaguet, Jordi M; Nofrarías, Miquel; Escribano, José M; Gómez-Sebastián, Silvia; Segalés, Joaquim; Rodríguez, Fernando

    2008-12-01

    The main aim of the present study was to describe new methods for the identification of antibodies against the PCV2 capsid (Cap) and replicase (Rep) proteins in pig sera. Specifically, two new indirect enzyme-linked immunosorbent assays (ELISA) were developed based on recombinant PCV2 Cap (rCap) and Rep/Rep' (rRep) proteins expressed in baculovirus and produced in Trichoplusia ni insect larvae. Both assays were validated by testing serum samples in a longitudinal study of 107 animals with different clinico-pathological features of PCV2 infection: pigs with postweaning multisystemic wasting syndrome (PMWS), wasted pigs without a diagnosis of PMWS and healthy animals. Longitudinal antibody profiles indicated that healthy animals had significantly higher anti-Cap and anti-Rep antibody levels than the rest of the animal groups at 11 weeks of age. Moreover, PMWS affected pigs could be distinguished from the rest of the pig groups by their lower anti-Rep antibody levels at 11 weeks of age and at necropsy. The results demonstrate the potential of these two ELISAs for large-scale serological studies. This study represents the first longitudinal study of the induction of anti-Cap and anti-Rep antibodies in farms affected by PMWS, from 1 week of age until the occurrence of disease. PMID:18773923

  11. Development and validation of an immunogold chromatographic test for on-farm detection of PRRSV.

    Science.gov (United States)

    Zhou, Sheng-hua; Cui, Shang-jin; Chen, Chang-mu; Zhang, Fan-chao; Li, Jun; Zhou, Shun; Oh, Jin-Sik

    2009-09-01

    An immunochromatographic test strip was developed to detect porcine reproductive and respiratory syndrome virus (PRRSV). The test uses two gold-labeled monoclonal antibodies: D5 against recombinant nucleocapsid protein (rN) and E9 against recombinant M protein (rM). In the test, PRRSV binds to a mixture of D5 and E9 labeled with colloidal gold; the complexes move through a membrane and are captured by rabbit anti-rM and anti-rN antibodies at a test line, producing a reddish-purple band because of the increased concentration of gold. Unbound monoclonal antibodies move past the test line to be captured by goat anti-mouse antibodies, producing a band at a control line. In samples without PRRSV or with low virus concentration, a band appears only at the control line. A crossover-trial demonstrated that the test strip was highly specific for PRRSV. The test strip detection limit was between 7.8x10(3) and 1.6x10(4) TCID(50)/ml. Analysis of 100 clinical samples indicated that the sensitivity, specificity, and accuracy of the immunochromatographic test strip relative to reverse transcription polymerase chain reaction (RT-PCR) were 97.0, 93.9, and 96.0%, respectively. Because the test is simple and rapid, it can be used by an unskilled person to detect PRRSV in the field. PMID:19427332

  12. Development of a Computerized Visual Search Test

    Science.gov (United States)

    Reid, Denise; Babani, Harsha; Jon, Eugenia

    2009-01-01

    Visual attention and visual search are the features of visual perception, essential for attending and scanning one's environment while engaging in daily occupations. This study describes the development of a novel web-based test of visual search. The development information including the format of the test will be described. The test was designed…

  13. The development of an Enzyme Linked Immunosorbent Assay for detecting Injectious laryngotrachitis viral antibodies in chicken serum

    OpenAIRE

    2002-01-01

    The aim of this study was to develop an Enzyme-linked immunosorbent assay (ELISA) for detection of antibody against gallid herpes virus, the causal agent of infectious laryngotracheitis (ILT) in chicken. Its application in experimental chicken under laboratory condition was also evaluated. Results showed that ELISA for ILT was developed successfully with sensitivity and specificity was 98% and 97,14% respectively. The ELISA was able to determine the dynamic of antibodies respond in experiment...

  14. Protective levels of canine distemper virus antibody in an urban dog population using plaque reduction neutralization test

    Directory of Open Access Journals (Sweden)

    O.I. Oyedele

    2004-11-01

    Full Text Available Blood samples from 50 dogs were collected at three veterinary clinics in Ibadan and Abuja, Nigeria and the serum from each sample was evaluated serologically for neutralizing antibodies against canine distemper virus (CDV by the highly sensitive plaque reduction (PRN neutralization assay. Thirteen dogs had plaque reduction neutralization titres of 0-100, seven had titres of 100-1 000 while 30 had titres ranging from 1 000-6 000. The PRN titres of vaccinated dogs were found to be significantly higher than unvaccinated dogs. The widespread use of the highly reproducible PRN test for the evaluation of antibody response to CDV may be very important in the generation of international CDV positive serum standards that should help to improve pre-and post-vaccination testing of dogs worldwide.

  15. Ability to develop broadly neutralizing HIV-1 antibodies is not restricted by the germline immunoglobulin gene repertoire1

    Science.gov (United States)

    Scheepers, Cathrine; Shrestha, Ram K.; Lambson, Bronwen E.; Jackson, Katherine J. L.; Wright, Imogen A.; Naicker, Dshanta; Goosen, Mark; Berrie, Leigh; Ismail, Arshad; Garrett, Nigel; Karim, Quarraisha Abdool; Karim, Salim S. Abdool; Moore, Penny L.; Travers, Simon A.; Morris, Lynn

    2015-01-01

    The human immunoglobulin repertoire is vast, producing billions of unique antibodies from a limited number of germline immunoglobulin genes. The immunoglobulin heavy chain variable region (IGHV) is central to antigen binding and is comprised of 48 functional genes. Here we analyzed whether HIV-1 infected individuals who develop broadly neutralizing antibodies show a distinctive germline IGHV profile. Using both 454 and Illumina technologies we sequenced the IGHV repertoire of 28 HIV-infected South African women from the Center for the AIDS Programme of Research in South African (CAPRISA) 002 and 004 cohorts, 13 of whom developed broadly neutralizing antibodies. Of the 259 IGHV alleles identified in this study, approximately half were not found in the International Immunogenetics Database (IMGT). This included 85 entirely novel alleles and 38 alleles that matched rearranged sequences in non-IMGT databases. Analysis of the rearranged H chain V region genes of monoclonal antibodies isolated from 7 of the CAPRISA women and previously isolated broadly neutralizing antibodies from other donors provided evidence that at least 8 novel or non-IMGT alleles contributed to functional antibodies. Importantly, we found that despite a wide range in the number of IGHV alleles in each individual, including alleles used by known broadly neutralizing antibodies, there were no significant differences in germline IGHV repertoires between individuals who do and do not develop broadly neutralizing antibodies. This study reports novel IGHV repertoires and highlights the importance of a fully comprehensive immunoglobulin database for germline gene usage prediction. Furthermore, these data suggest a lack of genetic bias in broadly neutralizing antibody development in HIV-1 infection, with implications for HIV vaccine design. PMID:25825450

  16. Waste form development/test

    International Nuclear Information System (INIS)

    The main objective of this study is to investigate new solidification agents relative to their potential application to wastes generated by advanced high volume reduction technologies, e.g., incinerator ash, dry solids, and ion exchange resins. Candidate materials selected for the solidification of these wastes include a modified sulfur cement and low-density polyethylene, neither of which are currently employed commerically for the solidification of low-level waste (LLW). As both the modified sulfur cement and the polyethylene are thermoplastic materials, a heated screw type extruder is utilized in the production of waste form samples for testing and evaluation. In this regard, work is being conducted to determine the range of conditions under which these solidification agents can be satisfactorily applied to the specific LLW streams and to provide information relevant to operating parameters and process control

  17. Automated serological technique with special emphasis on a solid phase test for red cell antibody detection in routine blood banking

    OpenAIRE

    Sallander, Suzanne

    1999-01-01

    Automated serological techniques for erythrocyte antigen typing and antibody screening are presented and evaluated in a larger number of samples and throughout routine processing. Both techniques are microplate-adapted with computerised sample identification, sample and reagent dispensing, and interpretation of results. The method described for typing of the RBC antigens K, Fya, and C, c, E, e compared well to the manual haernagglutination test. The concurrence was >= 99.4 %...

  18. Evaluation of capillary zone electrophoresis for charge heterogeneity testing of monoclonal antibodies.

    Science.gov (United States)

    Moritz, Bernd; Schnaible, Volker; Kiessig, Steffen; Heyne, Andrea; Wild, Markus; Finkler, Christof; Christians, Stefan; Mueller, Kerstin; Zhang, Li; Furuya, Kenji; Hassel, Marc; Hamm, Melissa; Rustandi, Richard; He, Yan; Solano, Oscar Salas; Whitmore, Colin; Park, Sung Ae; Hansen, Dietmar; Santos, Marcia; Lies, Mark

    2015-03-01

    with a 30cm capillary which demonstrates that an increased stability indicating potential can be combined with the increased separation velocity and high throughput capability of a shorter capillary. Separation can be performed in as little as approx. 3min allowing high throughput applications. The intercompany study delivered precise results without explicit training of the participating labs in the method prior to the study (standard deviations in the range of 1%). It was demonstrated that CZE is an alternative platform technology for the charge heterogeneity testing of antibodies in the pharmaceutical industry. PMID:25637812

  19. Test Marketing in New Product Development

    Science.gov (United States)

    Klompmaker, Jay E.; And Others

    1976-01-01

    Discusses the role of test marketing in new product development, based on interviews with marketing executives. Attempts to clarify when a test market should be done, what its aims should be, and how it should be used. (JG)

  20. Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis

    Directory of Open Access Journals (Sweden)

    Luciana Pereira Silva

    2003-07-01

    Full Text Available The present study was conducted to evaluate the frequency of antigenic components recognized by serum IgG antibodies in Western blotting (WB using a Strongyloides ratti larval extract for the diagnosis of human strongyloidiasis. In addition, the WB results were compared to the enzyme-linked immunosorbent assay (ELISA and the indirect immunofluorescence antibody test (IFAT results. Serum samples of 180 individuals were analyzed (80 with strongyloidiasis, 60 with other intestinal parasitoses, and 40 healthy individuals. S. ratti was obtained from fecal culture of experimentally infected Rattus rattus. For IFAT, S. ratti larvae were used as antigen and S. ratti larval antigenic extracts were employed in WB and ELISA. Eleven S. ratti antigenic components were predominantly recognized by IgG antibodies in sera of patients with strongyloidiasis. There was a positive concordance for the three tests in 87.5% of the cases of strongyloidiasis. The negative concordance in the three tests was 94% and 97.5%, in patients with other intestinal parasitoses and healthy individuals, respectively. In cases of positive ELISA and negative IFAT results, diagnosis could be confirmed by WB. ELISA, IFAT, and WB using S. ratti antigens showed a high rate of sensitivity and specificity. In conclusion, WB using S. ratti larval extract was able to recognize 11 immunodominant antigenic components, showing to be a useful tool to define the diagnosis in cases of equivocal serology.

  1. Antibody titers against swine influenza subtypes determined by the hemagglutination inhibition test are highly dependent on the strain

    DEFF Research Database (Denmark)

    Trebbien, Ramona; Nielsen, Jens; Bøtner, Anette;

    diagnostic and epidemiological point of view it is crucial to clarify whether the immunological response to one subtype protects against infection with the other subtype. The hemagglutination inhibition test (HI-test) has been used widely to determine the presence of antibodies in serum against influenza......In Denmark there are three circulating strains of swine influenza H1N1, H1N2 and H3N2. The H1N2 is different from the H1N2 subtypes circulating in other European countries. The Danish subtype is a reassortment between the two Danish circulating swine influenza subtypes H1N1 and H3N2. From a...... viruses. In the present study the HI-test was used to determine antibody response from experimental infected pigs. The aim of the study was to evaluate the antibody response against the new Danish influenza subtype H1N2 (H1N2dk) and to examine the level of crossprotection/reaction between the two...

  2. Development and utilization of camelid VHH antibodies from alpaca for 2,2',4,4'-tetrabrominated diphenyl ether detection.

    Science.gov (United States)

    Bever, Candace R S; Majkova, Zuzana; Radhakrishnan, Rajeswaran; Suni, Ian; McCoy, Mark; Wang, Yanru; Dechant, Julie; Gee, Shirley; Hammock, Bruce D

    2014-08-01

    An antibody-based analytical method for the detection of a chemical flame retardant using antibody fragments isolated from an alpaca has been developed. One specific chemical flame retardant congener, 2,2',4,4'-tetrabrominated diphenyl ether (BDE-47), is often the major poly-BDE (PBDE) congener present in human and environmental samples and that which is the most frequently detected. An alpaca was immunized with a surrogate of BDE-47 covalently attached to a carrier protein. The resulting mRNA coding for the variable domain of heavy-chain antibodies (VHH) were isolated, transcribed to cDNA, and cloned into a phagemid vector for phage display library construction. Selection of VHHs recognizing BDE-47 was achieved by panning under carefully modified conditions. The assay sensitivity for detecting BDE-47 was down to the part-per-billion (microgram per liter) level. Cross-reactivity analyses confirmed that this method was highly selective for BDE-47 and selected hydroxylated metabolites. When exposed to elevated temperatures, the camelid VHH antibodies retained more reactivity than a polyclonal antibody developed to the same target analyte. The use of this VHH antibody reagent immobilized onto a Au electrode for impedance biosensing demonstrates the increased versatility of VHH antibodies. PMID:25005746

  3. Development of diagnostic RI test method for antiglutamic acid decarboxylase (GAD) in SMS and IDDM patients

    Energy Technology Data Exchange (ETDEWEB)

    Ota, Mitsuhiro; Ota, Kiyoe; Nishimura, Masataka; Ma Jie; Obayashi, Hiroshi; Saida, Takahiko [Utano National Hospital, Kyoto (Japan)

    2000-02-01

    Western blotting with antigens purified using its specific antibody bound column has demonstrated that patients with Stiff-man syndrome (SMS) and insulin-dependent diabetic mellitus (IDDM) were both positive for anti-GAD antibody. Further, anti-GAD antibodies from various animal brains were characterized using GAD 65 and GAD 67 peptide antibody. The antibody against the anti-N-terminal peptide inhibited the enzyme activity of GAD, suggesting that the active site of GAD might exist in the N-terminal region. Development of a new detection method for anti-GAD antibody was attempted and the amount of GAD protein bound to protein G resin was determined based on the activity to release {sup 14}CO{sub 2} from {sup 14}C glutamic acid. In addition, solid-phase RIA method was developed using GAD purified by the anti-peptide antibody affinity column. The positive detection rate for GAD antibody was 39% for the enzymatic method and 56% for the solid-phase RIA method. To develop a further sensitive detection method for GAD antibody, construction of recombinant GAD was attempted and two GAD65s different in molecular size were constructed using pMal-c vector. Thus obtained antibodies against anti-N-terminal peptides were separately responded to GAD65 and GAD67 isoforms in the rat, mouse and bovine brains, whereas the carboxy-terminal antibodies were reactive to both isoforms together. Therefore, it became possible to make purification of GAD65 and GAD67 by the use of the two N-terminal peptide antibodies. Further, it became possible to purify GAD as a mixture of both isoforms. However, the yield of purification using anti-affinity column was still unsatisfactory ( several percent) and the GAD preparation obtained had little activity. The positive detection by the solid-phase RIA method was 50% for SMS patients and 56% for IDDM ones, indicating that this method was superior to the previous enzyme method. The protein A method in which labeled human recombinant GAD65 was used to

  4. Development of diagnostic RI test method for antiglutamic acid decarboxylase (GAD) in SMS and IDDM patients

    International Nuclear Information System (INIS)

    Western blotting with antigens purified using its specific antibody bound column has demonstrated that patients with Stiff-man syndrome (SMS) and insulin-dependent diabetic mellitus (IDDM) were both positive for anti-GAD antibody. Further, anti-GAD antibodies from various animal brains were characterized using GAD 65 and GAD 67 peptide antibody. The antibody against the anti-N-terminal peptide inhibited the enzyme activity of GAD, suggesting that the active site of GAD might exist in the N-terminal region. Development of a new detection method for anti-GAD antibody was attempted and the amount of GAD protein bound to protein G resin was determined based on the activity to release 14CO2 from 14C glutamic acid. In addition, solid-phase RIA method was developed using GAD purified by the anti-peptide antibody affinity column. The positive detection rate for GAD antibody was 39% for the enzymatic method and 56% for the solid-phase RIA method. To develop a further sensitive detection method for GAD antibody, construction of recombinant GAD was attempted and two GAD65s different in molecular size were constructed using pMal-c vector. Thus obtained antibodies against anti-N-terminal peptides were separately responded to GAD65 and GAD67 isoforms in the rat, mouse and bovine brains, whereas the carboxy-terminal antibodies were reactive to both isoforms together. Therefore, it became possible to make purification of GAD65 and GAD67 by the use of the two N-terminal peptide antibodies. Further, it became possible to purify GAD as a mixture of both isoforms. However, the yield of purification using anti-affinity column was still unsatisfactory ( several percent) and the GAD preparation obtained had little activity. The positive detection by the solid-phase RIA method was 50% for SMS patients and 56% for IDDM ones, indicating that this method was superior to the previous enzyme method. The protein A method in which labeled human recombinant GAD65 was used to precipitate 125-I GAD

  5. Software Testing Process in Agile Development

    OpenAIRE

    Malik, Ahsan Nawaz & Kashif Masood

    2008-01-01

    Software testing is the most important process to verify the quality of a product. Software testing in Agile development is very complex and controversial issue in literature and industry. Different people have different views about software testing in Agile methods, because most of Agile methods do not focus much on software testing activities. Agile strongly focus on the close customer collaboration, short iterations and frequent deliveries. But when it comes to software testing, then it is...

  6. Development of fuel test loop in HANARO

    International Nuclear Information System (INIS)

    KAERI have developed a fuel test loop facility to conduct the fuel irradiation test at HANARO. Maximum 3 pins of fuel can be tested in the IR-1 irradiation hole of HANARO under commercial power plant operating conditions. The integral system performance test with mock-up fuels under a high temperature is being performed. The FTL will be used for an advanced fuel irradiation test and could maximize the usage of HANARO. (author)

  7. Development of 2 types of competitive enzyme-linked immunosorbent assay for detecting antibodies to the rinderpest virus using a monoclonal antibody for a specific region of the hemagglutinin protein.

    Science.gov (United States)

    Khamehchian, S; Madani, R; Rasaee, M J; Golchinfar, F; Kargar, R

    2007-06-01

    A competitive enzyme-linked immunosorbent assay (C-ELISA) has been developed and standardized for the detection of antibodies to the rinderpest virus (RPV) in sera from cattle, sheep, and goats. The test is specific for rinderpest because it does not detect antibodies to peste-des-petits-ruminants virus (PPRV). The test depends on the ability of the monoclonal antibody (MAb) directed against the hemagglutinin (H) protein of RPV to compete with the binding of RPV antibodies in the positive serum to the H protein of this virus. This MAb recognized a region from amino acids 575 to 583 on the H protein of RPV that is unique to the RPV H protein and is not present on the hemagglutinin-neuraminidase protein of PPRV. Another C-ELISA (peptide C-ELISA) was set up using this specific region as an antigen. A threshold value of 64.4% inhibition was established for the RPV C-ELISA, with 90 known RPV-negative and 30 RPV-positive serum samples. Using common serum samples, a cutoff value of 43.0% inhibition for the peptide C-ELISA was established. Based on statistical analysis, the overall sensitivity and specificity of the RPV C-ELISA, relative to those of a commercial kit, were found to be 90.00% and 103.33%, respectively. However, the sensitivity and specificity of the peptide C-ELISA were found to be 180.00% and 73.33%, respectively. Although a common MAb in 2 new C-ELISA systems was used, variation in their percent inhibition, due to the use of different antigens, was observed. Taking into consideration the difference in percent inhibition of the 2 described assays and the commercial kit (50%), it was found that the RPV C-ELISA and the peptide C-ELISA are more specific and sensitive tools than the commercial kit for assessing herd immune status and for epidemiologic surveillance. PMID:17668032

  8. Testing Infrastructure for Operating System Kernel Development

    DEFF Research Database (Denmark)

    Walter, Maxwell; Karlsson, Sven

    2014-01-01

    Testing is an important part of system development, and to test effectively we require knowledge of the internal state of the system under test. Testing an operating system kernel is a challenge as it is the operating system that typically provides access to this internal state information. Multi......-core kernels pose an even greater challenge due to concurrency and their shared kernel state. In this paper, we present a testing framework that addresses these challenges by running the operating system in a virtual machine, and using virtual machine introspection to both communicate with the kernel...... and obtain information about the system. We have also developed an in-kernel testing API that we can use to develop a suite of unit tests in the kernel. We are using our framework for for the development of our own multi-core research kernel....

  9. The development of an Enzyme Linked Immunosorbent Assay for detecting Injectious laryngotrachitis viral antibodies in chicken serum

    Directory of Open Access Journals (Sweden)

    Risa Indriani

    2002-06-01

    Full Text Available The aim of this study was to develop an Enzyme-linked immunosorbent assay (ELISA for detection of antibody against gallid herpes virus, the causal agent of infectious laryngotracheitis (ILT in chicken. Its application in experimental chicken under laboratory condition was also evaluated. Results showed that ELISA for ILT was developed successfully with sensitivity and specificity was 98% and 97,14% respectively. The ELISA was able to determine the dynamic of antibodies respond in experimental chickens following vaccination and artificial infection with ILT virus. It was concluded that this ELISA offers a simple, sensitive and specific antibody assay for detection of antibodies against ILT virus in chickens arising from vaccination or infection.

  10. Diagnosis and Clinical Virology of Lassa Fever as Evaluated by Enzyme-Linked Immunosorbent Assay, Indirect Fluorescent-Antibody Test, and Virus Isolation

    Science.gov (United States)

    Bausch, D. G.; Rollin, P. E.; Demby, A. H.; Coulibaly, M.; Kanu, J.; Conteh, A. S.; Wagoner, K. D.; McMullan, L. K.; Bowen, M. D.; Peters, C. J.; Ksiazek, T. G.

    2000-01-01

    The Lassa virus (an arenavirus) is found in West Africa, where it sometimes causes a severe hemorrhagic illness called Lassa fever. Laboratory diagnosis has traditionally been by the indirect fluorescent-antibody (IFA) test. However, enzyme-linked immunosorbent assays (ELISAs) for Lassa virus antigen and immunoglobulin M (IgM) and G (IgG) antibodies have been developed that are thought to be more sensitive and specific. We compared ELISA and IFA testing on sera from 305 suspected cases of Lassa fever by using virus isolation with a positive reverse transcription-PCR (RT-PCR) test as the “gold standard.” Virus isolation and RT-PCR were positive on 50 (16%) of the 305 suspected cases. Taken together, Lassa virus antigen and IgM ELISAs were 88% (95% confidence interval [CI], 77 to 95%) sensitive and 90% (95% CI, 88 to 91%) specific for acute infection. Due to the stringent gold standard used, these likely represent underestimates. Diagnosis could often be made on a single serum specimen. Antigen detection was particularly useful in providing early diagnosis as well as prognostic information. Level of antigenemia varied inversely with survival. Detection by ELISA of IgG antibody early in the course of illness helped rule out acute Lassa virus infection. The presence of IFA during both acute and convalescent stages of infection, as well as significant interobserver variation in reading the slides, made interpretation difficult. However, the assay provided useful prognostic information, the presence of IFA early in the course of illness correlating with death. The high sensitivity and specificity, capability for early diagnosis, and prognostic value of the ELISAs make them the diagnostic tests of choice for the detection of Lassa fever. PMID:10878062

  11. Characterization and evaluation of monoclonal antibodies developed for typing influenza A and influenza B viruses.

    OpenAIRE

    Walls, H H; Harmon, M W; Slagle, J J; Stocksdale, C; Kendal, A P

    1986-01-01

    Monoclonal antibodies that are broadly reactive with influenza A or influenza B viruses were produced as stable reagents for typing influenza viruses. Monoclonal antibodies to influenza A were specific for either matrix protein or nucleoprotein. The antibodies to influenza B were specific for nucleoprotein or hemagglutinin protein. In an enzyme immunoassay procedure, influenza A antibodies detected H1N1, H2N2, and H3N2 influenza A virus strains collected between 1934 and 1984. Each of the inf...

  12. AngularJS test-driven development

    CERN Document Server

    Chaplin, Tim

    2015-01-01

    This book is for developers who want to learn about AngularJS development by applying testing techniques. You are assumed to have a basic knowledge and understanding of HTML, JavaScript, and AngularJS.

  13. The Utilization of a New Immunochromatographic Test in Detection of Helicobacter pylori Antibody from Maternal and Umbilical Cord Serum

    Directory of Open Access Journals (Sweden)

    Fu-Chen Kuo

    2014-01-01

    Full Text Available Background. Helicobacter pylori (H. pylori was linked with several extragastrointestinal diseases, including preeclampsia and intrauterine growth restriction of fetus. One of the signals which can be transferred from mother to fetus is the H. pylori IgG antibody. Aims. We utilized a commercial immunochromatographic kit to detect the antibody in maternal and cord serum. Methods. Three hundred and forty-six females were enrolled and the blood samples were collected on antenatal examination and on delivery. The maternal H. pylori infection was determined by stool H. pylori antigen test. Results. One hundred and five females (30.3% were H. pylori-infected, and the prevalence was higher in immigrants (43.5% than in Taiwanese (28.7%, P=0.058. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the kit were 77.1%, 88.0%, 73.6%, 89.8%, and 84.7%, respectively. This kit also had similar performance in cord serum. Comparing to the maternal result on delivery, this kit offered a consistent performance in antenatal maternal serum (kappa coefficient 0.92 and in cord serum (kappa coefficient 0.88. Conclusions. H. pylori IgG antibody can be transferred through the placenta into the fetal circulation. However, accuracy of the test kit needs to be evaluated before utilization in screening.

  14. Comparison between the Counter Immunoelectrophoresis Test and Mouse Neutralization Test for the Detection of Antibodies against Rabies Virus in Dog Sera

    Directory of Open Access Journals (Sweden)

    Luzia Helena Queiroz da Silva

    2002-03-01

    Full Text Available The detection of rabies antibodies is extremely valuable for epidemiological studies, determination of immune status in man, animals, and for the diagnosis of the disease. Several serological procedures have been described for this purpose. The present study reports a comparison between counterimmunoelectrophoresis test (CIET and mouse neutralization test (MNT in the detection of antibodies against rabies virus from 212 serum samples of vaccinated dogs. The agreement between both techniques was 79.7% and a significative association was demonstrated. The correlation coefficients between MNT and the CIET titers was determined considering 88 samples showing positive results in both techniques [CIET = 2 and MNT = 5 (0.13 IU/ml] and resulted r² = 0.7926 (p < 0.001. The performance of CIET system was technically simple, cheap and rapid, and thereby it could be useful for serological monitoring of dog vaccination campaigns as well as for individual analysis.

  15. Test Driven Development of Scientific Models

    Science.gov (United States)

    Clune, Thomas L.

    2014-01-01

    Test-Driven Development (TDD), a software development process that promises many advantages for developer productivity and software reliability, has become widely accepted among professional software engineers. As the name suggests, TDD practitioners alternate between writing short automated tests and producing code that passes those tests. Although this overly simplified description will undoubtedly sound prohibitively burdensome to many uninitiated developers, the advent of powerful unit-testing frameworks greatly reduces the effort required to produce and routinely execute suites of tests. By testimony, many developers find TDD to be addicting after only a few days of exposure, and find it unthinkable to return to previous practices.After a brief overview of the TDD process and my experience in applying the methodology for development activities at Goddard, I will delve more deeply into some of the challenges that are posed by numerical and scientific software as well as tools and implementation approaches that should address those challenges.

  16. Development and characterization of synthetic antibodies binding to the cystic fibrosis conductance regulator.

    Science.gov (United States)

    Gakhal, Amandeep K; Jensen, Timothy J; Bozoky, Zoltan; Roldan, Ariel; Lukacs, Gergely L; Forman-Kay, Julie; Riordan, John R; Sidhu, Sachdev S

    2016-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel in the apical surface of epithelial cells in the airway and gastrointestinal tract, and mutation of CFTR is the underlying cause of cystic fibrosis. However, the precise molecular details of the structure and function of CFTR in native and disease states remains elusive and cystic fibrosis researchers are hindered by a lack of high specificity, high affinity binding reagents for use in structural and biological studies. Here, we describe a panel of synthetic antigen-binding fragments (Fabs) isolated from a phage-displayed library that are specific for intracellular domains of CFTR that include the nucleotide-binding domains (NBD1 and NBD2), the R-region, and the regulatory insertion loop of NBD1. Binding assays performed under conditions that promote the native fold of the protein demonstrated that all Fabs recognized full-length CFTR. However, only the NBD1-specific Fab recognized denatured CFTR by western blot, suggesting a conformational epitope requirement for the other Fabs. Surface plasmon resonance experiments showed that the R-region Fab binds with high affinity to both the phosphorylated and unphosphorylated R-region. In addition, NMR analysis of bound versus unbound R-region revealed a distinct conformational effect upon Fab binding. We further defined residues involved with antibody recognition using an overlapping peptide array. In summary, we describe methodology complementary to previous hybridoma-based efforts to develop antibody reagents to CFTR, and introduce a synthetic antibody panel to aid structural and biological studies. PMID:27185291

  17. Integrated Language Definition Testing: Enabling Test-Driven Language Development

    NARCIS (Netherlands)

    Kats, L.C.L.; Vermaas, R.; Visser, E.

    2011-01-01

    The reliability of compilers, interpreters, and development environments for programming languages is essential for effective software development and maintenance. They are often tested only as an afterthought. Languages with a smaller scope, such as domain-specific languages, often remain untested.

  18. Development of Lateral-flow Immunoassay for WSSV with Polyclonal Antibodies Raised against Recombinant VP (19+28) Fusion Protein

    Institute of Scientific and Technical Information of China (English)

    Qing-yu CHENG; Xiao-lin MENG; Jin-ping XU; Wei LU; Jian WANG

    2007-01-01

    We have developed a sensitive and rapid lateral-flow immunoassay (LFIA) for WSSV,using colloidal gold as an indicator.The fusion protein,VP (19+28),was expressed in E.coli,purified and used to prepare polyclonal antibodies.The purified anti-VP (19+28) IgG were conjugated with colloidal gold.Unconjugated anti-VP (19+28) IgG and goat anti-rabbit IgG were immobilized on nitrocellulose membranes.After assembly,three groups (5 individual animals in each group) of shrimp samples were tested which included healthy,moribund and dead shrimps.For each group,three different tissues (body juices,gills and hepatopancreas) were tested at the same time.In parallel,all the samples were also analyzed using PCR for comparison.Out of 45 samples tested,30 were detected as positive while 15 were classified as negative.The results of LFIA correlate with those obtained by the PCR analysis,indicating that these two detection methods have the same efficacy in the limited number of samples tested in this preliminary study.

  19. Development of automatic weld strength testing machine

    International Nuclear Information System (INIS)

    In order to improve the testing process and accuracy so as to carry out all the manual works including documentation automatically and effortlessly, an automatic computerised strength testing machine with the latest state-of-art technology, including both the hardware and software was developed. The operator has to only submit the weld to the machine for testing and start the testing process merely by pressing a switch. This paper depicts the salient features of this machine

  20. Development of a multi-product leached protein A assay for bioprocess samples containing recombinant human monoclonal antibodies.

    Science.gov (United States)

    Ren, Diya; Darlucio, Maria R; Chou, Judy H

    2011-03-01

    The detection of low level of protein A leached from monoclonal antibody downstream purification process is often interfered by the presence of excess amount of product antibody. In order to prevent this interference, we developed a new multi-product leached protein A assay that used acidification to completely dissociate the IgG-ProteinA complex, followed by neutralization under selected condition to prevent re-formation of the IgG-ProteinA complex. The amount of protein A was then determined by a sandwich immunoassay using Meso Scale Discovery technology. The assay takes approximately 3h to complete for one 96-well plate of samples, and this has been successfully applied to samples containing different monoclonal antibody products examined so far. The data demonstrates that this assay is robust and efficient in determining leached protein A contamination during purification of recombinant monoclonal antibodies. PMID:21315722

  1. The acceptability of the introduction of a type specific herpes antibody screening test into a genitourinary medicine clinic in the United Kingdom

    OpenAIRE

    Mullan, H; Munday, P.

    2003-01-01

    Objective: To determine the uptake of a type specific herpes simplex antibody test if it were offered as part of routine screening in a genitourinary medicine clinic in a district general hospital in the United Kingdom.

  2. Coombs Antiglobulin Test Using Brucella abortus 99 as Antigen To Detect Incomplete Antibodies Induced by B. abortus RB51 Vaccine in Cattle

    OpenAIRE

    Ciuchini, Franco; Adone, Rosanna; Pasquali, Paolo

    2002-01-01

    This study showed that vaccination of cattle with Brucella abortus rough strain RB51 induces incomplete antibodies that can be detectable by a Coombs antiglobulin test using the B. abortus 99 smooth strain.

  3. Seroprevalence of Mycoplasma gallisepticum antibody by ELISA and serum plate agglutination test of laying chicken

    OpenAIRE

    Md. Zulfekar Ali; Md. Mostafizer Rahman; Shirin Sultana

    2015-01-01

    Aim: Mycoplasma gallisepticum (MG) is important avian pathogens responsible for chronic respiratory diseases of chicken and turkeys, which result in large economic loss for the poultry industry. The objectives of this study were determination of seroprevalence of MG antibody of commercial layer chicken at laying period in selected areas of Bangladesh. Materials and Methods: A total of 563 blood samples were collected randomly from selected commercial layer chickens at laying period during the...

  4. Analysis of speckled fluorescent antinuclear antibody test antisera using electrofocused nuclear antigens.

    OpenAIRE

    Okarma, T B; Krueger, J A; Holman, H. R.

    1982-01-01

    Antibodies to different components of the extractable nuclear antigen (ENA) have been thought to be serological markers for clinical subsets of rheumatic diseases. However, incomplete characterization and standardization of antigenic components such as ribonucleoprotein (RNP), Sm, and SS-B (Ha), and the multiplicity of autoantibodies produced by different patients have confounded correlations between autoantibody specificity and disease subsets. This study describes the preparative separation...

  5. Development of an in Vitro Potency Assay for Anti-anthrax Lethal Toxin Neutralizing Antibodies

    Directory of Open Access Journals (Sweden)

    Sjoerd Rijpkema

    2012-01-01

    Full Text Available Lethal toxin (LT of Bacillus anthracis reduces the production of a number of inflammatory mediators, including transcription factors, chemokines and cytokines in various human cell lines, leading to down-regulation of the host inflammatory response. Previously we showed that the reduction of interleukin-8 (IL-8 is a sensitive marker of LT-mediated intoxication in human neutrophil-like NB-4 cells and that IL-8 levels are restored to normality when therapeutic monoclonal antibodies (mAb with toxin-neutralising (TN activity are added. We used this information to develop cell-based assays that examine the effects of TN therapeutic mAbs designed to treat LT intoxication and here we extend these findings. We present an in vitro assay based on human endothelial cell line HUVEC jr2, which measures the TN activity of therapeutic anti-LT mAbs using IL-8 as a marker for intoxication. HUVEC jr2 cells have the advantage over NB-4 cells that they are adherent, do not require a differentiation step and can be used in a microtitre plate format and therefore can facilitate high throughput analysis. This human cell-based assay provides a valid alternative to the mouse macrophage assay as it is a more biologically relevant model of the effects of toxin-neutralising antibodies in human infection.

  6. Efficacy of serum samples stored on filter paper for the detection of antibody to Leptospira spp. by microagglutination test (MAT).

    Science.gov (United States)

    Blanco, R M; Romero, E C

    2012-12-14

    The aim of this study was to investigate the microagglutination test (MAT) results in serum samples dried on filter paper and stored at different temperatures during 1day, 7days, 30days and 1year to determine the stability of sera antibody against leptospires. Serum samples collected onto filter paper for the detection of leptospires antibody was compared with MAT in a study of 300 serum samples from patients with suspected leptospirosis. Among 300 fresh serum samples analyzed by MAT 156 (52%) were positive and 144 (48%) negative. All the negative fresh serum samples were negative when dried on filter paper (specificity 100%). The sensitivity of MAT performed on dried serum samples was 100%. Storage on filter paper at room temperature and at 4°C for 1 and 7days did not affect the MAT titers. For up to 7days, 98.72% of dried serum samples had titers identical to those of the corresponding serum samples, and 1.18% of dried serum samples showed 1 dilution of difference. After a storage period of one month a prozone phenomenon was observed. After a storage period of one year all serum samples were negative. Serum samples collected onto filter paper are a convenient source of antibodies for serological diagnosis and epidemiological surveys. PMID:22960422

  7. Evaluation of an immunochromatographic test for rapid and reliable serodiagnosis of human tularemia and detection of Francisella tularensis-specific antibodies in sera from different mammalian species.

    Science.gov (United States)

    Splettstoesser, W; Guglielmo-Viret, V; Seibold, E; Thullier, P

    2010-05-01

    Tularemia is a highly contagious infectious zoonosis caused by the bacterial agent Francisella tularensis. Serology is still considered to be a cornerstone in tularemia diagnosis due to the low sensitivity of bacterial culture and the lack of standardization in PCR methodology for the direct identification of the pathogen. We developed a novel immunochromatographic test (ICT) to efficiently detect F. tularensis-specific antibodies in sera from humans and other mammalian species (nonhuman primate, pig, and rabbit). This new tool requires none or minimal laboratory equipment, and the results are obtained within 15 min. When compared to the method of microagglutination, which was shown to be more specific than the enzyme-linked immunosorbent assay, the ICT had a sensitivity of 98.3% (58 positive sera were tested) and a specificity of 96.5% (58 negative sera were tested) on human sera. On animal sera, the overall sensitivity was 100% (22 positive sera were tested) and specificity was also 100% (70 negative sera were tested). This rapid test preferentially detects IgG antibodies that may occur early in the course of human tularemia, but further evaluation with human sera is important to prove that the ICT can be a valuable field test to support a presumptive diagnosis of tularemia. The ICT can also be a useful tool to monitor successful vaccination with subunit vaccines or live vaccine strains containing lipopolysaccharide (e.g., LVS) and to detect seropositive individuals or animals in outbreak situations or in the context of epidemiologic surveillance programs in areas of endemicity as recently recommended by the World Health Organization. PMID:20220165

  8. The diagnostic sensitivity of dengue rapid test assays is significantly enhanced by using a combined antigen and antibody testing approach.

    OpenAIRE

    Fry, Scott R.; Michelle Meyer; Semple, Matthew G.; Cameron P Simmons; Shamala Devi Sekaran; Huang, Johnny X.; Catriona McElnea; Chang-Yi Huang; Andrea Valks; Paul R. Young; Cooper, Matthew A.

    2011-01-01

    BACKGROUND: Serological tests for IgM and IgG are routinely used in clinical laboratories for the rapid diagnosis of dengue and can differentiate between primary and secondary infections. Dengue virus non-structural protein 1 (NS1) has been identified as an early marker for acute dengue, and is typically present between days 1-9 post-onset of illness but following seroconversion it can be difficult to detect in serum. AIMS: To evaluate the performance of a newly developed Panbio® Dengue Early...

  9. The Diagnostic Sensitivity of Dengue Rapid Test Assays Is Significantly Enhanced by Using a Combined Antigen and Antibody Testing Approach

    OpenAIRE

    Fry, Scott R.; Meyer, Michelle; Semple, Matthew G.; Cameron P. Simmons; Sekaran, Shamala Devi; Johnny X. Huang; McElnea, Catriona; Huang, Chang-Yi; Valks, Andrea; Young, Paul R.; Cooper, Matthew A.

    2011-01-01

    Background Serological tests for IgM and IgG are routinely used in clinical laboratories for the rapid diagnosis of dengue and can differentiate between primary and secondary infections. Dengue virus non-structural protein 1 (NS1) has been identified as an early marker for acute dengue, and is typically present between days 1–9 post-onset of illness but following seroconversion it can be difficult to detect in serum. Aims To evaluate the performance of a newly developed Panbio® Dengue Early R...

  10. Test and Behaviour Driven Development with Python

    CERN Document Server

    CERN. Geneva

    2016-01-01

    Experience has taught us that bugs are impossible to avoid when programming. Specially on continuous delivery processes where there are new versions that refactor or incorporate new modules to the project. Although, there are different tools which help us to ensure code quality by enabling developers to catch bugs while still in the development stage. In this talk, I will talk about Test-driven development(TDD) and Behaviour-Driven development (BDD) methodologies focused on web development. Also, I will present an overview of unit testing tools as Selenium or Behave, which help us to produce working software, with fewer bugs, quickly and consistently.

  11. The preparation of monoclonal antibodies against human proinsulin and the development of immunoradiometric assay

    International Nuclear Information System (INIS)

    To prepare monoclonal antibodies against human proinsulin (hPI) and to develop a sensitive and specific two-site immunoradiometric assay for human proinsulin. Monoclonal antibodies (McAbs) against human hPi were prepared by the routine method. Two of these McAbs were selected to develop a sensitive and specific two-site IRMA for serum hPI. We used this assay to measure the fasting and postprandial serum concentrations of PI in healthy subjects, patients with NIDDM and those with insulinoma. We obtained 4 mouse McAbs (2A6, 4A2, 4E10, 4H9) against hPI. McAb 4H9 probably binds to an epitope that is located at the junction of A chain or B chain of insulin with c-peptide and the three others bind to epitopes that are located on the A chain or B chain of insulin. The detection limit of the IRMA for hPI was 1.9 pmol/L and the assay showed no cross-reaction with insulin and C-peptide. The mean fasting concentrations of PI was 5.5 +- 3.3 pmol/L and increased to 17.3 +-6.4 pmol/L one hour after a glucose loading in healthy subjects. They were 9.4% +- 1.5 pmol/L and 31.8 +- 9.4 pmol/L in patients with NIDDM. In a patient with insulinoma, the fasting concentration of PI was 106 pmol/L and 57 pmol/L before and one week after operation respectively. Based on the hPI specific McAbs produced in our laboratory, we developed a sensitive and specific two-site IRMA for serum hPI. It will be useful in the diagnosis of insulinoma and in the elucidation of the physiological and pathophysiological significance of PI in human

  12. Correlation of serum antithyroid microsomal antibody and autologous serum skin test in patients with chronic idiopathic urticaria

    Directory of Open Access Journals (Sweden)

    Snehal Balvant Lunge

    2015-01-01

    Full Text Available Background: About 25-45% of patients of chronic urticaria (CU have been stated to have histamine releasing autoantibodies in their blood. The term autoimmune urticaria is increasingly being accepted for this subgroup of patients. Review of the literature suggests high autologous serum skin test (ASST positivity and presence of antithyroid microsomal antibodies in patients with autoimmune urticaria. Aims: To study prevalence of ASST positivity and antithyroid microsomal antibodies in chronic "idiopathic" urticaria and to study the correlation between the two parameters. Methods: All patients of chronic idiopathic urticaria satisfying inclusion/exclusion criteria were enrolled in the study after written informed consent. Patients of CU secondary to infections and infestations, physical urticaria including dermatographism, mastocytosis, urticarial vasculitis and those on treatment with immunosuppressive drugs for urticaria were excluded from the study. In all of these patients, complete blood count; ASST, serum T3/T4/thyroid stimulating hormone levels, antithyroid microsomal antibody (AMA levels were done. Statistical analysis was done by Chi-square test, Fisher exact test and Kappa statistics. Results: Study included 24 males and 26 females with mean age of 39.54 years. Majority of patients belonged to 20-40 years of age. Females showed more ASST positivity. A total of 12 out of 50 (24% patients showed positive ASST. A total of four out of 12 (33.33% had positive ASST and raised AMA levels. Conclusion: Only 25% of patients of chronic idiopathic urticaria had positive ASST. ASST and AMA levels were positively correlated in our study. Further studies are required to authenticate this association.

  13. Preliminary studies of Technetium-99m-labeled antimyosin monoclonal antibody: development of radiopharmaceutical for cardiac evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Carvalho, Guilherme Luiz de Castro; Spencer, Patrick Jack; Muramoto, Emiko; Araujo, Elaine Bortoleti de [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)]. E-mail: glcarval@ipen.br

    2007-07-01

    In the acute myocardium infarction, the myocytes cell membrane loses its integrity, allowing the influx of extracellular macromolecules such as circulating antibody into the damaged cell. Specific antibodies to cardiac myosin can therefore bind to the acutely necrotic myocyte, allowing the noninvasive localization and dimension of myocardial infarction. Because of its favorable physical characteristics, low cost, and ready availability, technetium-99m ({sup 99m}Tc) is the radionuclide of choice for scintigraphy. The purpose of this work was to study the labeling of the antimyosin monoclonal antibody with ({sup 99m}Tc for development of a radiopharmaceutical with high sensitivity and specificity used in the diagnostic of the myocardial infarction. The intact monoclonal antibody (IgG{sub 1}) was reduced by treatment with dithiothreitol (DTT) with the consequent generation of free thiol groups (- SH), responsible for the labeling of the antibody with ({sup 99m}Tc. The radiochemical yield was determined using Sephadex G-25 column (PD-10). The percentage of ({sup 99m}Tc-antibody was 90,06% and after purification procedure the radiochemical yield was > 98%. The biodistribution studies showed low uptake in the stomach and thyroid at different times (1, 4 e 24 hours) representing a small amount of unbounded ({sup 99m}Tc and a good stability of the purified ({sup 99m}Tc-antibody. The uptake in the normal heart was relatively low as expected. Based on these results, we concluded that the direct labeling procedure applied to the antimyosin monoclonal antibody allowed the easy preparation of the radiopharmaceutical with good stability to be used in the noninvasive diagnostic of the myocardial infarction. (author)

  14. Development of enhanced antibody response toward dual delivery of nano-adjuvant adsorbed human Enterovirus-71 vaccine encapsulated carrier.

    Science.gov (United States)

    Saeed, Mohamed I; Omar, Abdul Rahman; Hussein, Mohd Z; Elkhidir, Isam M; Sekawi, Zamberi

    2015-01-01

    This study introduces a new approach for enhancing immunity toward mucosal vaccines. HEV71 killed vaccine that is formulated with nanosize calcium phosphate adjuvant and encapsulated onto chitosan and alginate delivery carriers was examined for eliciting antibody responses in serum and saliva collected at weeks 0, 1, 3, 5, 7 and 9 for viral-specific IgA & IgG levels and viral neutralizing antibody titers. The antibody responses induced in rabbits by the different formulations delivered by a single (buccal) route were compared to those of dual immunization (intradermal / mucosal) and un-immunized control. Chitosan-loaded vaccine adjuvant induced elevated IgA antibody, while Alginate-adjuvant irreversible bonding sequestered the vaccine and markedly reduced immunogenicity. The induced mucosal and parenteral antibody profiles appeared in an inverse manner of enhanced mucosal IgA antibody accompanied by lower systemic IgG following a single oral immunization route. The combined intradermal and oral dual-immunized group developed an elevated salivary IgA, systemic IgG, and virus neutralizing response. A reduced salivary neutralizing antibody titer was observed and attributed to the continual secretion exchanges in saliva. Designing a successful mucosal delivery formulation needs to take into account the vaccine delivery site, dosage, adjuvant and carrier particle size, charge, and the reversibility of component interactions. The dual immunization seems superior and is a important approach for modulating the antibody response and boosting mucosal protection against HEV71 and similar pathogens based on their transmission mode, tissue tropism and shedding sites. Finally, the study has highlighted the significant role of dual immunization for simultaneous inducing and modulating the systemic and mucosal immune responses to EV71. PMID:26186664

  15. Lead Coolant Test Facility Development Workshop

    Energy Technology Data Exchange (ETDEWEB)

    Paul A. Demkowicz

    2005-06-01

    A workshop was held at the Idaho National Laboratory on May 25, 2005, to discuss the development of a next generation lead or lead-alloy coolant test facility. Attendees included representatives from the Generation IV lead-cooled fast reactor (LFR) program, Advanced Fuel Cycle Initiative, and several universities. Several participants gave presentations on coolant technology, existing experimental facilities for lead and lead-alloy research, the current LFR design concept, and a design by Argonne National Laboratory for an integral heavy liquid metal test facility. Discussions were focused on the critical research and development requirements for deployment of an LFR demonstration test reactor, the experimental scope of the proposed coolant test facility, a review of the Argonne National Laboratory test facility design, and a brief assessment of the necessary path forward and schedule for the initial stages of this development project. This report provides a summary of the presentations and roundtable discussions.

  16. Cost Savings Associated with Testing of Antibodies, Antigens, and Nucleic Acids for Diagnosis of Acute HIV Infection

    OpenAIRE

    Karris, Maile Y.; Anderson, Christy M.; Sheldon R. Morris; Smith, Davey M.; Little, Susan J.

    2012-01-01

    Efforts to identify all persons infected with HIV in the United States are driven by the hope that early diagnosis will lower risk behaviors and decrease HIV transmission. Identification of HIV-infected people earlier in the course of their infection with HIV antigen/antibody (Ag/Ab) combination assays (4th-generation HIV assays) should help achieve this goal. We compared HIV RNA nucleic acid test (NAT) results to the results of a 4th-generation Ag/Ab assay (Architect HIV Ag/Ab Combo [HIV Com...

  17. Review for the generalist: The antinuclear antibody test in children - When to use it and what to do with a positive titer

    Directory of Open Access Journals (Sweden)

    Sailer-Hoeck Michaela

    2010-10-01

    Full Text Available Abstract The antinuclear antibody test (ANA is a much overused test in pediatrics. The ANA does have a role in serologic testing but it should be a very limited one. It is often ordered as a screening test for rheumatic illnesses in a primary care setting. However, since it has low specificity and sensitivity for most rheumatic and musculoskeletal illnesses in children, it should not be ordered as a screening test for non-specific complaints such as musculoskeletal pain. It should only be used as a diagnostic test for children with probable Systemic Lupus Erythematosus (SLE or Mixed Connective Tissue Disease, (MCTD and other possible overlap-like illnesses. Such children should have developed definite signs and symptoms of a disease before the ANA is ordered. This review presents data supporting these conclusions and a review of the ANA literature in adults and children. By limiting ANA testing, primary care providers can avoid needless venipuncture pain, unnecessary referrals, extra medical expenses, and most importantly, significant parental anxieties. It is best not to do the ANA test in most children but if it ordered and is positive in a low titer (

  18. Testing automation tools for secure software development

    OpenAIRE

    Eatinger, Christopher J.

    2007-01-01

    Software testing is a crucial step in the development of any software system, large or small. Testing can reveal the presence of logic errors and other flaws in the code that could cripple the system's effectiveness. Many flaws common in software today can also be exploited to breach the security of the system on which the software is running. These flaws can be subtle and difficult to find. Frequently it takes a combination of multiple events to bring them out. Traditional testing techni...

  19. Development and testing of a VTOL UAV

    OpenAIRE

    Cibula, Andrew Lee.

    1995-01-01

    The purpose of this project was to develop a testing platform to prepare a Vertical Takeoff and Landing Unmanned Air Vehicle (VTOL UAV) for fully independent flight tests. Other preparations for flight included extensive engine thrust and endurance testing to fully evaluate the capabilities of the engine used. Also, redesign of the fuel system allowed more efficient use of the fuel onboard. Commands for thrust and steering data were transmitted to the VTOL UAV via an RF uplink while UAV attit...

  20. Validation of antibody ELISA for use in combination with parasitological tests in monitoring trypanosomosis control programmes

    International Nuclear Information System (INIS)

    The available techniques for trypanosomosis diagnosis, surveillance and control have failed to provide an effective control of the disease when used in isolation. However, when used in combination the techniques might provide a better alternative for a sustainable disease control strategy. Antigen-detection ELISA was validated and established at ADRI through support of the Joint FAO/IAEA Division, but still required perfection in order to meet the requirements for which it was intended (detection of current infection and species specificity). The primary objective of the present study was to evaluate the trypanosomal antibody-detection ELISA (Ab-ELISA) kit produced by the International Atomic Energy Agency (IAEA) Vienna, Austria, to supplement conventional diagnostic techniques in trypanosomosis surveillance and control. Three-hundred sixty cattle from selected herds in the Tanga region of Tanzania were bled to obtain buffy-coat and serum samples in a study of the prevalence of trypanosomosis using parasitological and serological techniques. The prevalence of trypanosomosis, by means of parasitological techniques, in Korogwe, Muheza, Tanga and Pangani districts of Tanga region, varied from 0-18.2%. The IAEA Ab-ELISA kit, showed a 97.2% specificity for Trypanosoma congolense and T. vivax antibodies, at a cut-off point of 30.0% positivity (PP), out of 160 sera from a trypanosomosis-negative population; and a 94.5% sensitivity for the same antibodies and cut-off point, out of 200 sera from trypanosomosis-buffy-coat-positive samples. Packed red cell volume (PCV) values for a trypanosomosis-negative population were much higher than those from cattle from trypanosomosis-endemic areas: 77.5% of samples from a disease-free area showed PCV values higher than 30.0, while only 15% of the samples from endemic areas were above 30.0. (author)

  1. /sup 99m/Tc radiolabelling and quality control tests of anti-melanoma monoclonal antibodies and F(ab')/sub 2/ fragments for immunoscintigraphy

    International Nuclear Information System (INIS)

    Tumour radioimmunodetection was first developed by using radiolabelled polyclonal antibodies, raised in goats against tumour associated antigens (TAA). The availability of monoclonal antibodies to TAA has definitely contributed to more extensive in vivo use of radiolabelled antibodies. However, many factors are involved in tumour radioimmunolocalization, related either to the antibody and radioisotope features or to the natural history of the tumour itself. The experimental protocol developed by the authors allows a full evaluation of the properties of a particular MoAb.This paper illustrates the work done with on a particular set of monoclonal antibodies, raised against human melanoma associated antigens, with the aim of visualizing primary and metastatic lesions in melanoma patients

  2. Development and Characterization of a Monoclonal Antibody against Ochratoxin B and Its Application in ELISA

    Directory of Open Access Journals (Sweden)

    Alexandra H. Heussner

    2010-06-01

    Full Text Available A monoclonal antibody specific to ochratoxin B (OTB was employed for the development of an indirect competitive OTB-ELISA. The optimized OTB-ELISA resulted in a limit of detection (LOD for OTB of 3 µg/L (8 nM, a limit of quantification (LOQ of 3.7 µg/L (10 nM, and a 50% inhibitory concentration (IC50 of 150 nM. Due to very low cross-reactivity to OTA (2.7% and structurally related molecules (0%, this OTB-ELISA was found to be suitable to detect OTB with excellent precision in different matrices, i.e., beer, coffee and wine. Therefore, this OTB-ELISA will allow screening of OTB in food and feed products.

  3. Testbed For Aerothermal Test Technique Development Project

    Data.gov (United States)

    National Aeronautics and Space Administration — It is proposed that a very low cost wind tunnel could be developed at JSC to provide engineers with the ability to directly run small tests focused on improving...

  4. Developing New Testing Methods for Nanosatellites Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Thermal modeling and Test plan to be carried out and developed by Goddard Space Flight Center. This project will be done in collaboration with partners at MIT and...

  5. Development of a Danish speech intelligibility test

    DEFF Research Database (Denmark)

    Nielsen, Jens Bo; Dau, Torsten

    2009-01-01

    Abstract A Danish speech intelligibility test for assessing the speech recognition threshold in noise (SRTN) has been developed. The test consists of 180 sentences distributed in 18 phonetically balanced lists. The sentences are based on an open word-set and represent everyday language. The sente...

  6. Software Testing Platform Development and Implementation

    OpenAIRE

    Burian, Vojtěch

    2012-01-01

    The quality is probably the most significant property of a successful software product. As experience with many software projects has already shown, leaving out testing and quality management from software development process can result in vast and critical customer issues, which usually invoke additional expenses for the software production company. In the course of time, software testing as a discipline has therefore seized an important position among other software development activities. ...

  7. The Historical Development of Animal Toxicity Testing

    OpenAIRE

    Gertler, N.

    1997-01-01

    This paper traces the historical development of animal toxicity testing, from its ancient origins through the period of standardization following World War II. It explores the roots of toxicity testing in physiology and experimental medicine, drug development, and the detection and identification of poisons. The discussion then turns to the shift in focus from acute to chronic toxicity which occurred around the turn of the century. The controversy over the potential toxicity of preservatives ...

  8. Monoclonal antibodies.

    Science.gov (United States)

    2009-01-01

    The ability to produce and exploit monoclonal antibodies (mAbs) has revolutionized many areas of biological sciences. The unique property of an mAb is that it is a single species of immunoglobulin (IG) molecule. This means that the specificity of the interaction of the paratopes on the IG, with the epitopes on an antigenic target, is the same on every molecule. This property can be used to great benefit in immunoassays to provide tests of defined specificity and sensitivity, which improve the possibilities of standardization. The performance of assays can often be determined relating the actual weight of antibody (hence the number of molecules) to the activity. Often the production of an mAb against a specific epitope is the only way that biological entities can be differentiated. This chapter outlines the areas involving the development of assays based on mAbs. The problems involved address include the physical aspects of mAbs and how they may affect assay design and also the implications of results based on monospecific reagents. Often these are not fully understood, leading to assays that are less than satisfactory, which does not justify the relatively high cost of preparing and screening of mAbs. There are many textbooks and reviews dealing with the preparation of mAbs, the principles involved, and various purification and manipulative methods for the preparation of fragments and conjugation. There has been little general information attempting to summarize the best approaches to assay design using mAbs. Much time can be wasted through bad planning, and this is particularly relevant to mAbs. A proper understanding of some basic principles is essential. It is beyond the scope of this chapter to discuss all aspects, but major areas are highlighted. PMID:19219589

  9. Incidence of Neisseria gonorrhoeae isolates negative by Syva direct fluorescent-antibody test but positive by Gen-Probe accuprobe test in a sexually transmitted disease clinic population.

    Science.gov (United States)

    Beebe, J L; Rau, M P; Flageolle, S; Calhoon, B; Knapp, J S

    1993-09-01

    To determine the accuracy of the Syva (Palo Alto, Calif.) direct fluorescent-antibody (DFA) test in comparison with the Gen-Probe (San Diego, Calif.) Accuprobe culture confirmation test, we tested 395 isolates of Neisseria gonorrhoeae from cultures obtained from patients attending a sexually transmitted disease clinic from 1 July 1991 through 30 June 1992. All isolates were tested for DFA reactivity with a polyclonal reagent (Difco Laboratories, Detroit, Mich.) and a monoclonal reagent (Syva, Inc., direct specimen test) and for specific molecular probe reactivity by the Gen-Probe Accuprobe culture confirmation test for N. gonorrhoeae. The 395 isolates gave positive results for the Gen-Probe culture confirmation test and the Difco polyclonal direct specimen test. However, 18 (4.6%) of the isolates were negative for N. gonorrhoeae by the Syva DFA test. With the exception of six beta-lactamase-positive isolates, all isolates that were negative by Syva DFA were sensitive to penicillin, tetracycline, spectinomycin, and ceftriaxone by disk-diffusion susceptibility testing. Auxotyping and serotyping studies indicated that strains negative by Syva DFA consisted of several variants. The frequency of N. gonorrhoeae isolates showing negative results by Syva DFA in this patient population ranged from 0 to 11.5%/month. Laboratories using only the Syva DFA test for confirmation of N. gonorrhoeae may incur a significant risk of misidentification. PMID:8408585

  10. Bispecific antibodies.

    Science.gov (United States)

    Kontermann, Roland E; Brinkmann, Ulrich

    2015-07-01

    Bispecific antibodies (bsAbs) combine specificities of two antibodies and simultaneously address different antigens or epitopes. BsAbs with 'two-target' functionality can interfere with multiple surface receptors or ligands associated, for example with cancer, proliferation or inflammatory processes. BsAbs can also place targets into close proximity, either to support protein complex formation on one cell, or to trigger contacts between cells. Examples of 'forced-connection' functionalities are bsAbs that support protein complexation in the clotting cascade, or tumor-targeted immune cell recruiters and/or activators. Following years of research and development (R&D), the first bsAb was approved in 2009. Another bsAb entered the market in December 2014 and several more are in clinical trials. Here, we describe the potentials of bsAbs to become the next wave of antibody-based therapies, focusing on molecules in clinical development. PMID:25728220

  11. Transport Test Problems for Hybrid Methods Development

    Energy Technology Data Exchange (ETDEWEB)

    Shaver, Mark W.; Miller, Erin A.; Wittman, Richard S.; McDonald, Benjamin S.

    2011-12-28

    This report presents 9 test problems to guide testing and development of hybrid calculations for the ADVANTG code at ORNL. These test cases can be used for comparing different types of radiation transport calculations, as well as for guiding the development of variance reduction methods. Cases are drawn primarily from existing or previous calculations with a preference for cases which include experimental data, or otherwise have results with a high level of confidence, are non-sensitive, and represent problem sets of interest to NA-22.

  12. Developing high-quality mouse monoclonal antibodies for neuroscience research - approaches, perspectives and opportunities.

    Science.gov (United States)

    Gong, Belvin; Murray, Karl D; Trimmer, James S

    2016-09-25

    High-quality antibodies (Abs) are critical to neuroscience research, as they remain the primary affinity proteomics reagent used to label and capture endogenously expressed protein targets in the nervous system. As in other fields, neuroscientists are frequently confronted with inaccurate and irreproducible Ab-based results and/or reporting. The UC Davis/NIH NeuroMab Facility was created with the mission of addressing the unmet need for high-quality Abs in neuroscience research by applying a unique approach to generate and validate mouse monoclonal antibodies (mAbs) optimized for use against mammalian brain (i.e., NeuroMabs). Here we describe our methodology of multi-step mAb screening focused on identifying mAbs exhibiting efficacy and specificity in labeling mammalian brain samples. We provide examples from NeuroMab screens, and from the subsequent specialized validation of those selected as NeuroMabs. We highlight the particular challenges and considerations of determining specificity for brain immunolabeling. We also describe why our emphasis on extensive validation of large numbers of candidates by immunoblotting and immunohistochemistry against brain samples is essential for identifying those that exhibit efficacy and specificity in those applications to become NeuroMabs. We describe the special attention given to candidates with less common non-IgG1 IgG subclasses that can facilitate simultaneous multiplex labeling with subclass-specific secondary antibodies. We detail our recent use of recombinant cloning of NeuroMabs as a method to archive all NeuroMabs, to unambiguously define NeuroMabs at the DNA sequence level, and to re-engineer IgG1 NeuroMabs to less common IgG subclasses to facilitate their use in multiplex labeling. Finally, we provide suggestions to facilitate Ab development and use, as to design, execution and interpretation of Ab-based neuroscience experiments. Reproducibility in neuroscience research will improve with enhanced Ab validation

  13. Application of GP5 protein to develop monoclonal antibody against porcine reproductive and respiratory syndrome virus.

    Science.gov (United States)

    Tian, Hong; Cheng, Yan; Wu, Jin-yang; He, Jian-hui; Shang, You-jun; Liu, Xiang-tao

    2011-08-01

    In this study, a panel of monoclonal antibodies (mAbs) against Porcine reproductive and respiratory syndrome virus(PRRSV), named as 8C9 and4B4, were produced by fusing SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with the PRRSV (TCID(50)=5.5), screened by the indirect ELISA and subjected to several limiting dilutions. mAbs were then identified by biological characterization. Among the two fusion cell strains, 8C9 belonged to the IgG1 subclass and 4B4 belonged to the IgG2a subclass. The titers in cell culture supernatant and abdomen liquor reached to 1:10(4)and 1:10(5), respectively. The specificity test indicated that the two cells had specific reactions for the PRRSV and GP5 protein respectively, and no reaction with Classical swine fever virus (CSFV) or Swine vesicular disease virus (SVDV). The molecular weights of the heavy chain and light chain were about 45.0 kDa and 25.0 kDa, respectively. In neutralization activity tests, the results showed that the prepared mAb 4B4 can protect 50% of cells with no CPE in dilution up to 1:512, but mAB 8C9 has no neutralization activities to PRRSV. PMID:21847758

  14. Application of GP5 Protein to Develop Monoclonal Antibody against Porcine Reproductive and Respiratory Syndrome Virus

    Institute of Scientific and Technical Information of China (English)

    Hong Tian; Yan Cheng; Jin-yang Wu; Jian-hui He; You-jun Shang; Xiang-tao Liu

    2011-01-01

    In this study,a panel of monoclonal antibodies (mAbs) against Porcine reproductive and respiratory syndrome virus(PRRSV),named as 8C9 and4B4,were produced by fusing SP2/0 myeloma cells and spleen cells of BALB/c mice immunized with the PRRSV (TCID50=5.5),screened by the indirect ELISA and subjected to several limiting dilutions.mAbs were then identified by biological characterization.Among the two fusion cell strains,8C9 belonged to the IgG1 subclass and 4B4 belonged to the IgG2a subclass.The titers in cell culture supernatant and abdomen liquor reached to 1:104and 1:105,respectively.The specificity test indicated that the two cells had specific reactions for the PRRSV and GP5 protein respectively,and no reaction with Classical swine fever virus (CSFV) or Swine vesicular disease virus (SVDV).The molecular weights of the heavy chain and light chain were about 45.0 kDa and 25.0 kDa,respectively.In neutralization activity tests,the results showed that the prepared mAb 4B4 can protect 50% of cells with no CPE in dilution up to 1:512,but mAB 8C9 has no neutralization activities to PRRSV.

  15. Approach to a case of multiple irregular red cell antibodies in a liver transplant recipient: Need for developing competence

    Directory of Open Access Journals (Sweden)

    Ravi C Dara

    2015-01-01

    Full Text Available Liver transplant procedure acts as a challenge for transfusion services in terms of specialized blood components, serologic problems, and immunologic effects of transfusion. Red cell alloimmunization in patients awaiting a liver transplant complicate the process by undue delay or unavailability of compatible red blood cell units. Compatible blood units can be provided by well-equipped immunohematology laboratory, which has expertise in resolving these serological problems. This report illustrates resolution of a case with multiple alloantibodies using standard techniques, particularly rare antisera. Our case re-emphasizes the need for universal antibody screening in all patients as part of pretransfusion testing, which helps to identify atypical antibodies and plan for appropriate transfusion support well in time. We recommend that the centers, especially the ones that perform complex procedures like solid organ transplants and hematological transplants should have the necessary immunohematological reagents including rare antisera to resolve complex cases of multiple antibodies as illustrated in this case.

  16. Development and evaluation of a pseudovirus-luciferase assay for rapid and quantitative detection of neutralizing antibodies against enterovirus 71.

    Directory of Open Access Journals (Sweden)

    Xing Wu

    Full Text Available The level of neutralizing antibodies (NtAb induced by vaccine inoculation is an important endpoint to evaluate the efficacy of EV71 vaccine. In order to evaluate the efficacy of EV71 vaccine, here, we reported the development of a novel pseudovirus system expression firefly luciferase (PVLA for the quantitative measurement of NtAb. We first evaluated and validated the sensitivity and specificity of the PVLA method. A total of 326 serum samples from an epidemiological survey and 144 serum specimens from 3 clinical trials of EV71 vaccines were used, and the level of each specimen's neutralizing antibodies (NtAb was measured in parallel using both the conventional CPE-based and PVLA-based assay. Against the standard neutralization assay based on the inhibition of the cytopathic effect (CPE, the sensitivity and specificity of the PVLA method are 98% and 96%, respectively. Then, we tested the potential interference of NtAb against hepatitis A virus, Polio-I, Polio-II, and Polio-III standard antisera (WHO and goat anti-G10/CA16 serum, the PVLA based assay showed no cross-reactivity with NtAb against other specific sera. Importantly, unlike CPE based method, no live replication-competent EV71 is used during the measurement. Taken together, PVLA is a rapid and specific assay with higher sensitivity and accuracy. It could serve as a valuable tool in assessing the efficacy of EV71 vaccines in clinical trials and disease surveillance in epidemiology studies.

  17. Development of a New Indirect ELISA Method for Detection of Anti-Tuberculosis Antibodies in Human Serum

    Directory of Open Access Journals (Sweden)

    Mohammad Morad-Farajollahi

    2015-10-01

    Full Text Available Background: Tuberculosis is a crucial health problem. Establishing a rapid, reliable and still inexpensive diagnostic method for tuberculosis seems to be substantial in developing countries where TB has very high incidence rate. Methods:  An Indirect  Enzyme-linked  immunosorbent  Assay (ELISA  was established to detect serum antibodies against Mycobacterium  tuberculosis. Three kinds of antigens were used to prepare the solid phase for antibody as- say including: purified protein derivative (PPD, M. tuberculosis Bacilli, and Mycobacterium  bovis Bacillus  Calmette  Guerin  (BCG.  Sera  of two main following groups were investigated in this study: sera samples from smear- positive, culture-positive and Tuberculin Skin Test-positive TB patients and sera samples from smear-negative, culture negative and TST-negative healthy individuals.Results: Among the antigens used, BCG produced higher sensitivity and spe-cificity in the assay. With PPD as the solid phase, higher sensitivity, but low- er specificity was observed in comparison with BCG. Both, low response and noise (non-specific binding were observed with TB bacilli as the solid phase in the assay.Conclusion: Using BCG solid phase system in this method resulted in highersensitivity in comparison to single antigen solid phase systems. In addition, we were able to circumvent  the problem of non-specific  bindings in more popular multi-antigenic solid systems such as PPD. By using this new indi- rect ELISA, a rapid, reliable and still inexpensive diagnosis of tuberculosis might be possible. Although,  further investigations  are required to confirm our result.

  18. Development of a simple method for the immobilization of anti-thyroxine antibody on polystyrene tubes for use in the measurement of total thyroxine in serum

    International Nuclear Information System (INIS)

    We describe a simple method for the immobilisation of anti-thyroxine antibody on to the surface of polystyrene tubes and a simple assay format for the quantitative estimation of total thyroxine in serum. The immobilisation of anti-thyroxine antibody was achieved through passive adsorption of normal rabbit gamma globulin and anti-rabbit antibody raised in goat, as immune bridges. This procedure ensured minimum utilisation of primary and secondary antibody as neat sera without precipitation or affinity purification. The developed assay system using these antibody coated tubes covers a range of 0-240 ng/mL of thyroxine with intra and inter assay variations of less than 10 %. (author)

  19. Experiments toward the development of a radioimmunoassay for the detection of serum antibodies for the respiratory syncytial virus

    International Nuclear Information System (INIS)

    In order to detect an infection by the respiratory syncytial virus (RSV) quickly and safely, a radioimmunassay (RIA) should be developed. Various antigen preparations were compared to one another. The immune serums used in the RIA came from guinea pigs with a RSV antibody titer of up to 320 in the complement binding reaction. A number of observations lead to the discussion of the possibility of the formation (incomplete) of cross-reactive antibodies between virus and host cell. This hypothesis could be well supported through references in the literature. Under the assumption of the existence of cross-reactive antibodies, a further model of the pathogenesis of the RSV illness allows itself to be developed, which could be preceived as an illness with autoimmune components. With this model the varying courses of this disease in different age groups can be easily explained. (orig.)

  20. Comparison between results of virus neutralization test and those of two ELISAs when screening for antibodies to pseudorabies virus in Thailand

    International Nuclear Information System (INIS)

    The virus neutralization (VN) test and two enzyme linked immunosorbent assays (blocking and indirect ELISAs) were used to detect antibodies to pseudorabies virus on serum samples from 1000 pigs from the central part of Thailand. Using the VN test as standard, the blocking and indirect ELISAs showed respectively 95.12% and 99.37% relative sensitivity and 92.0% and 93.5% relative specificity. The two ELISAs were both considered as practical alternatives to the VN test. However, the indirect ELISA was the more suitable test for the routine screening for antibodies to pseudorabies virus in Thailand. (author). 22 refs, 1 fig., 3 tabs

  1. Development of two murine antibodies against Neospora caninum using phage display technology and application on the detection of N. caninum.

    Directory of Open Access Journals (Sweden)

    Jinhua Dong

    Full Text Available Neosporosis, caused by an intracellular parasite, Neospora caninum, is an infectious disease primarily of cattle and dogs. It occurs worldwide and causes huge damages to dairy farms. In this study, we immunized mice with recombinant surface-associated protein 1 of N. caninum (rNcSAG1 and developed two novel monoclonal antibodies, A10 and H3, against NcSAG1 using phage-display technology. Both clones bound to purified rNcSAG1 and the half maximal inhibitory concentrations of A10 and H3 are 50 and 72 nM of rNcSAG1, respectively. In immunofluorescence assays, both A10 and H3 Fabs bound to N. caninum parasites. Direct detection of N. caninum parasites was developed firstly using an enzyme-linked immunosorbent assay (ELISA with A10 and H3. Binding of A10 and H3 antibodies to rNcSAG1 was also inhibited by some certain anti-N. caninum antibodies in the neosporosis-positive cattle sera, suggesting they might bind to the same epitopes of NcSAG1 with those anti-N. caninum antibodies of bovine. These antibodies were demonstrated to have a potential for monitoring the N. caninum parasites in a dairy farm, which may lead to protect livestock from parasite-infection.

  2. Development of electrochemical immunosensors based on different serum antibody immobilization methods for detection of Japanese encephalitis virus

    Science.gov (United States)

    Tran, Quang Huy; Hanh Nguyen, Thi Hong; Mai, Anh Tuan; Thuy Nguyen, Thi; Khue Vu, Quang; Nga Phan, Thi

    2012-03-01

    This paper describes the development of electrochemical immunosensors based on human serum antibodies with different immobilization methods for detection of Japanese encephalitis virus (JEV). Human serum containing anti-JEV antibodies was used to immobilize onto the surface of silanized interdigitated electrodes by four methods: direct adsorption (APTES-serum), covalent binding with a cross linker of glutaraldehyde (APTES-GA-serum), covalent binding with a cross linker of glutaraldehyde combined with anti-human IgG (APTES-GA-anti-HIgG-serum) and covalent binding with a cross linker of glutaraldehyde combined with a bioaffinity of protein A (APTES-GA-PrA-serum). Atomic force microscopy was used to verify surface characteristics of the interdigitated electrodes before and after treatment with serum antibodies. The output signal of the immunosensors was measured by the change of conductivity resulting from the specific binding of JEV antigens and serum antibodies immobilized on the electrodes, with the help of horseradish peroxidase (HRP)-labeled secondary antibody against JEV. The results showed that the APTES-GA-PrA-serum method provided the highest signal of the electrochemical immunosensor for detection of JEV antigens, with the linear range from 25 ng ml‑1 to 1 μg ml‑1, and the limit of detection was about 10 ng ml‑1. This study shows a potential development of novel electrochemical immunosensors applied for virus detection in clinical samples in case of possible outbreaks.

  3. Development of electrochemical immunosensors based on different serum antibody immobilization methods for detection of Japanese encephalitis virus

    International Nuclear Information System (INIS)

    This paper describes the development of electrochemical immunosensors based on human serum antibodies with different immobilization methods for detection of Japanese encephalitis virus (JEV). Human serum containing anti-JEV antibodies was used to immobilize onto the surface of silanized interdigitated electrodes by four methods: direct adsorption (APTES-serum), covalent binding with a cross linker of glutaraldehyde (APTES-GA-serum), covalent binding with a cross linker of glutaraldehyde combined with anti-human IgG (APTES-GA-anti-HIgG-serum) and covalent binding with a cross linker of glutaraldehyde combined with a bioaffinity of protein A (APTES-GA-PrA-serum). Atomic force microscopy was used to verify surface characteristics of the interdigitated electrodes before and after treatment with serum antibodies. The output signal of the immunosensors was measured by the change of conductivity resulting from the specific binding of JEV antigens and serum antibodies immobilized on the electrodes, with the help of horseradish peroxidase (HRP)-labeled secondary antibody against JEV. The results showed that the APTES-GA-PrA-serum method provided the highest signal of the electrochemical immunosensor for detection of JEV antigens, with the linear range from 25 ng ml−1 to 1 μg ml−1, and the limit of detection was about 10 ng ml−1. This study shows a potential development of novel electrochemical immunosensors applied for virus detection in clinical samples in case of possible outbreaks

  4. Diagnostic test accuracy of anti-glycopeptidolipid-core IgA antibodies for Mycobacterium avium complex pulmonary disease: systematic review and meta-analysis

    OpenAIRE

    Yuji Shibata; Nobuyuki Horita; Masaki Yamamoto; Toshinori Tsukahara; Hideyuki Nagakura; Ken Tashiro; Hiroki Watanabe; Kenjiro Nagai; Kentaro Nakashima; Ryota Ushio; Misako Ikeda; Atsuya Narita; Akinori Kanai; Takashi Sato; Takeshi Kaneko

    2016-01-01

    Currently, an anti-glycopeptidolipid (GPL)-core IgA antibody assay kit for diagnosing Mycobacterium avium complex (MAC) is commercially available. We conducted this systematic review and meta-analysis to reveal the precise diagnostic accuracy of anti-GPL-core IgA antibodies for MAC pulmonary disease (MAC-PD). We systematically searched reports that could provide data for both sensitivity and specificity by anti-GPL-core IgA antibody for clinically diagnosed MAC-PD. Diagnostic test accuracy wa...

  5. Development of a Fully Human Anti-PDGFRβ Antibody That Suppresses Growth of Human Tumor Xenografts and Enhances Antitumor Activity of an Anti-VEGFR2 Antibody

    Directory of Open Access Journals (Sweden)

    Juqun Shen

    2009-06-01

    Full Text Available Platelet-derived growth factor receptor β (PDGFRβ is upregulated in most of solid tumors. It is expressed by pericytes/smooth muscle cells, fibroblast, macrophage, and certain tumor cells. Several PDGF receptor-related antagonists are being developed as potential antitumor agents and have demonstrated promising antitumor activity in both preclinical and clinical settings. Here, we produced a fully human neutralizing antibody, IMC-2C5, directed against PDGFRβ from an antibody phage display library. IMC-2C5 binds to both human and mouse PDGFRβ and blocks PDGF-B from binding to the receptor. IMC-2C5 also blocks ligand-stimulated activation of PDGFRβ and downstream signaling molecules in tumor cells. In animal studies, IMC-2C5 significantly delayed the growth of OVCAR-8 and NCI-H460 human tumor xenografts in nude mice but failed to show antitumor activities in OVCAR-5 and Caki-1 xenografts. Our results indicate that the antitumor efficacy of IMC-2C5 is primarily due to its effects on tumor stroma, rather than on tumor cells directly. Combination of IMC-2C5 and DC101, an anti-mouse vascular endothelial growth factor receptor 2 antibody, resulted in significantly enhanced antitumor activity in BxPC-3, NCI-H460, and HCT-116 xenografts, compared with DC101 alone, and the trend of additive effects to DC101 treatment in several other tumor models. ELISA analysis of NCI-H460 tumor homogenates showed that IMC-2C5 attenuated protein level of vascular endothelial growth factor and basic fibroblast growth factor elevated by DC101 treatment. Finally, IMC-2C5 showed a trend of additive effects when combined with DC101/chemotherapy in MIA-PaCa-2 and NCI-H460 models. Taken together, these results lend great support to the use of PDGFRβ antagonists in combination with other antiangiogenic agents in the treatment of a broad range of human cancers.

  6. Thyroid Antibodies

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Thyroid Antibodies Share this page: Was this page helpful? Also known as: Thyroid Autoantibodies; Antithyroid Antibodies; Antimicrosomal Antibody; Thyroid Microsomal Antibody; ...

  7. Development and Characterization of Monoclonal Antibodies and Aptamers Against Major Antigens of Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Specific antibodies, available in unlimited quantities, have not been produced against Mycobacterium avium subsp. paratuberculosis, the bacterium that causes Johne’s disease (JD). To fill this gap in JD research, monoclonal antibodies (mAbs) against M. avium subsp. paratuberculosis were produced fr...

  8. Development of recombinant antibody technology for application in plant pathogen diagnosis

    NARCIS (Netherlands)

    Griep, R.A.

    1999-01-01

    This thesis describes the applicability of the novel phage display technique to select plant-pathogen-specific monoclonal antibodies (MAbs) from combinatorial antibody libraries. The retrieved MAbs are so specific that they can be used as diagnostic tools in sensitive immunoassays for the detection

  9. Development of Monoclonal Antibodies Which Specifically Recognize Entamoeba histolytica in Preserved Stool Samples

    OpenAIRE

    Yau, Yvonne C. W.; Crandall, Ian; kain, kevin c.

    2001-01-01

    We report the generation of monoclonal antibodies against a recombinant 170-kDa subunit of the Gal or GalNAc lectin of Entamoeba histolytica that specifically recognize E. histolytica but not Entamoeba dispar in preserved stool samples. These antibodies do not cross-react with other bowel protozoa, including Entamoeba coli, Giardia lamblia, and Dientamoeba fragilis.

  10. Progress in FMIT test assembly development

    International Nuclear Information System (INIS)

    Research and development supporting the completed design of the Fusion Materials Irradiation Test (FMIT) Facility is continuing at the Hanford Engineering Development Laboratory (HEDL) in Richland, Washington. The FMIT, a deuteron accelerator based (d + Li) neutron source, will produce an intense flux of high energy neutrons for use in radiation damage studies of fusion reactor materials. The most intense flux magnitude of greater than 1015 n/cm2-s is located close to the neutron producing lithium target and is distributed within a volume about the size of an American football. The conceptual design and development of FMIT experiments called Test Assemblies has progressed over the past five years in parallel with the design of the FMIT. The paper will describe the recent accomplishments made in developing test assemblies appropriate for use in the limited volume close to the FMIT target where high neutron flux and heating rates and the associated spacial gradients significantly impact design considerations

  11. [Effect of erythrocyte preserved for different lengths of time on anti-D antibody identification with three blood matching tests].

    Science.gov (United States)

    Xiao, Rui-Qing; Lin, Wu-Cun; Xu, Dan; Zeng, Jie; Wu, Jian-Jun; Zhao, Shu-Ming

    2003-10-01

    The specificity of the antigens and length of preservation time of erythrocytes are the interfering factors in blood group serological tests. In order to clarify the influence of preservation time of erythrocytes on the blood matching test, the titers of anti-D antibody were detected with papain method, BioVue cross matching card and DianaGel cross matching card in 7 series of panel red blood cells preserved for various length of time (0 to 9 months). The results showed that the titer of micro-column gel test (DianaGel card) was one tube higher than that of column agglutinating test (BioVue card). The titer of erythrocytes preserved for 9 months was as high as 256 tested by DianaGel card, but it was only 2 by papain method in the same anti-serum. It is suggested that there was no obvious difference between the results of micro-column gel test and column agglutinating test, and titer of papain method was the lowest. PMID:14575550

  12. Development of monoclonal antibodies against parathyroid hormone: genetic control of the immune response to human PTH

    International Nuclear Information System (INIS)

    Seventeen monocloanl antibodies against the aminoterminal portion of parathyroid hormone (PTH) were generated by using BALB/c mouse for immunization fully biologically active synthetic human PTH-(1-34) and bovine PTH-(1-84) as immunogens, monoclonal antibody methods, and a solid-phase screening assay. Isotypic analysis of these monoclonal antibodies was performed using affinity purified goat antimouse immunoglobulins specific for IgG heavy chains and μ(IgM). All antibodies were IgM as evidenced by 40 times greater than background activity when 25,000 cpm of 125I-labelled goat anti-mouse IgM was used as second antibody in a radioimmunoassay

  13. Sensitivity and specificity of typhoid fever rapid antibody tests for laboratory diagnosis at two sub-Saharan African sites

    Directory of Open Access Journals (Sweden)

    Karen H Keddy

    2011-09-01

    Full Text Available OBJECTIVE: To evaluate three commercial typhoid rapid antibody tests for Salmonella Typhi antibodies in patients suspected of having typhoid fever in Mpumalanga, South Africa, and Moshi, United Republic of Tanzania. METHODS: The diagnostic accuracy of Cromotest® (semiquantitative slide agglutination and single tube Widal test,TUBEX®and Typhidot® was assessed against that of blood culture. Performance was modelled for scenarios with pretest probabilities of 5% and 50%. FINDINGS: In total 92 patients enrolled: 53 (57.6% from South Africa and 39 (42.4% from the United Republic of Tanzania. Salmonella Typhi was isolated from the blood of 28 (30.4% patients. The semiquantitative slide agglutination and single-tube Widal tests had positive predictive values (PPVs of 25.0% (95% confidence interval, CI: 0.6-80.6 and 20.0% (95% CI: 2.5-55.6, respectively. The newer typhoid rapid antibody tests had comparable PPVs: TUBEX®, 54.1% (95% CI: 36.9-70.5; Typhidot® IgM, 56.7% (95% CI: 37.4-74.5; and Typhidot® IgG, 54.3% (95% CI: 36.6-71.2. For a pretest probability of 5%, PPVs were: TUBEX®, 11.0% (95% CI: 6.6-17.9; Typhidot® IgM, 9.1% (95% CI: 5.8-14.0; and Typhidot® IgG, 11.0% (6.3-18.4. For a pretest probability of 50%, PPVs were: TUBEX®, 70.2% (95% CI: 57.3-80.5; Typhidot® IgM, 65.6% (95% CI: 54.0-75.6; and Typhidot® IgG, 70.0% (95% CI: 56.0-81.1. CONCLUSION: Semiquantitative slide agglutination and single-tube Widal tests performed poorly. TUBEX® and Typhidot® may be suitable when pretest probability is high and blood cultures are unavailable, but their performance does not justify deployment in routine care settings in sub-Saharan Africa.

  14. Autologous HIV-1 neutralizing antibodies: emergence of neutralization-resistant escape virus and subsequent development of escape virus neutralizing antibodies

    DEFF Research Database (Denmark)

    Arendrup, M; Nielsen, C; Hansen, J E;

    1992-01-01

    The capacity of consecutive human sera to neutralize sequentially obtained autologous virus isolates was studied. HIV-1 was isolated three times over a 48-164-week period from three individuals immediately after seroconversion and from two individuals in later stages of infection. Development of ...

  15. Selection of Recombinant Human Antibodies.

    Science.gov (United States)

    Tomszak, Florian; Weber, Susanne; Zantow, Jonas; Schirrmann, Thomas; Hust, Michael; Frenzel, André

    2016-01-01

    Since the development of therapeutic antibodies the demand of recombinant human antibodies is steadily increasing. Traditionally, therapeutic antibodies were generated by immunization of rat or mice, the generation of hybridoma clones, cloning of the antibody genes and subsequent humanization and engineering of the lead candidates. In the last few years, techniques were developed that use transgenic animals with a human antibody gene repertoire. Here, modern recombinant DNA technologies can be combined with well established immunization and hybridoma technologies to generate already affinity maturated human antibodies. An alternative are in vitro technologies which enabled the generation of fully human antibodies from antibody gene libraries that even exceed the human antibody repertoire. Specific antibodies can be isolated from these libraries in a very short time and therefore reduce the development time of an antibody drug at a very early stage.In this review, we describe different technologies that are currently used for the in vitro and in vivo generation of human antibodies. PMID:27236551

  16. DTU PMU Laboratory Development - Testing and Validation

    DEFF Research Database (Denmark)

    Garcia-Valle, Rodrigo; Yang, Guang-Ya; Martin, Kenneth E.;

    2010-01-01

    This is a report of the results of phasor measurement unit (PMU) laboratory development and testing done at the Centre for Electric Technology (CET), Technical University of Denmark (DTU). Analysis of the PMU performance first required the development of tools to convert the DTU PMU data into IEEE...... standard, and the validation is done for the DTU-PMU via a validated commercial PMU. The commercial PMU has been tested from the authors' previous efforts, where the response can be expected to follow known patterns and provide confirmation about the test system to confirm the design and settings. In a...... nutshell, having 2 PMUs that observe same signals provides validation of the operation and flags questionable results with more certainty. Moreover, the performance and accuracy of the DTU-PMU is tested acquiring good and precise results, when compared with a commercial phasor measurement device, PMU-1....

  17. Evaluation of Multiplex-Based Antibody Testing for Use in Large-Scale Surveillance for Yaws: a Comparative Study.

    Science.gov (United States)

    Cooley, Gretchen M; Mitja, Oriol; Goodhew, Brook; Pillay, Allan; Lammie, Patrick J; Castro, Arnold; Moses, Penias; Chen, Cheng; Ye, Tun; Ballard, Ronald; Martin, Diana L

    2016-05-01

    WHO has targeted yaws for global eradication by 2020. The program goals are to interrupt the transmission in countries where yaws is endemic and to certify countries as yaws free where yaws was endemic in the past. No new rapid plasmin reagin (RPR) seroreactivity in young children is required for certification of elimination at a country level. We sought to evaluate whether antibody responses to specific treponemal antigens measured in a high-throughput multiplex bead array (MBA) assay differentiate past versus current infection and whether a nontreponemal lipoidal antigen test can be incorporated into the MBA. Serum and dried blood spot specimens collected for yaws surveillance projects in Ghana, Vanuatu, and Papua New Guinea (PNG) were run on MBA to measure antibodies against recombinant p17 (rp17) and treponemal membrane protein A (TmpA) treponemal antigens. Results were compared to standard treponemal laboratory (TPPA or TPHA [TPP(H)A]) and quantitative RPR test data. Of 589 specimens, 241 were TPP(H)A(+)/RPR(+), 88 were TPP(H)A(+)/RPR(-), 6 were TPP(H)A(-)/RPR(+), and 254 were negative for both tests. Compared to TPP(H)A, reactive concordance of rp17 was 93.7%, while reactive concordance of TmpA was only 81.9%. TmpA-specific reactivity showed good correlation with RPR titers (R(2) = 0.41; P < 0.0001). IgG responses to the lipoidal antigen used in RPR testing (cardiolipin) were not detected in the MBA. Our results suggest that TmpA can be used as a treponemal antigen marker for recent or active infection and potentially replace RPR in a high-throughput multiplex tool for large-scale yaws surveillance. PMID:26962086

  18. Evaluation of Multiplex-Based Antibody Testing for Use in Large-Scale Surveillance for Yaws: a Comparative Study

    Science.gov (United States)

    Cooley, Gretchen M.; Mitja, Oriol; Goodhew, Brook; Pillay, Allan; Lammie, Patrick J.; Castro, Arnold; Moses, Penias; Chen, Cheng; Ye, Tun; Ballard, Ronald

    2016-01-01

    WHO has targeted yaws for global eradication by 2020. The program goals are to interrupt the transmission in countries where yaws is endemic and to certify countries as yaws free where yaws was endemic in the past. No new rapid plasmin reagin (RPR) seroreactivity in young children is required for certification of elimination at a country level. We sought to evaluate whether antibody responses to specific treponemal antigens measured in a high-throughput multiplex bead array (MBA) assay differentiate past versus current infection and whether a nontreponemal lipoidal antigen test can be incorporated into the MBA. Serum and dried blood spot specimens collected for yaws surveillance projects in Ghana, Vanuatu, and Papua New Guinea (PNG) were run on MBA to measure antibodies against recombinant p17 (rp17) and treponemal membrane protein A (TmpA) treponemal antigens. Results were compared to standard treponemal laboratory (TPPA or TPHA [TPP(H)A]) and quantitative RPR test data. Of 589 specimens, 241 were TPP(H)A+/RPR+, 88 were TPP(H)A+/RPR−, 6 were TPP(H)A−/RPR+, and 254 were negative for both tests. Compared to TPP(H)A, reactive concordance of rp17 was 93.7%, while reactive concordance of TmpA was only 81.9%. TmpA-specific reactivity showed good correlation with RPR titers (R2 = 0.41; P < 0.0001). IgG responses to the lipoidal antigen used in RPR testing (cardiolipin) were not detected in the MBA. Our results suggest that TmpA can be used as a treponemal antigen marker for recent or active infection and potentially replace RPR in a high-throughput multiplex tool for large-scale yaws surveillance. PMID:26962086

  19. Development of single chain variable fragment (scFv) antibodies against surface proteins of 'Ca. Liberibacter asiaticus'.

    Science.gov (United States)

    Yuan, Qing; Jordan, Ramon; Brlansky, Ronald H; Minenkova, Olga; Hartung, John

    2016-03-01

    'Candidatus Liberibacter asiaticus' is the causal agent of citrus huanglongbing, the most serious disease of citrus worldwide. We have developed and applied immunization and affinity screening methods to develop a primary library of recombinant single chain variable fragment (scFv) antibodies in an M13 vector, pKM19. The antibody population is enriched for antibodies that bind antigens of 'Ca. Liberibacter asiaticus'. The primary library has more than 10(7) unique antibodies and the genes that encode them. We have screened this library for antibodies that bind to specifically-chosen proteins that are present on the surface of 'Ca. Liberibacter asiaticus'. These proteins were used as targets for affinity-based selection of scFvs that bind to the major outer membrane protein, OmpA; the polysaccharide capsule protein KpsF; a protein component of the type IV pilus (CapF); and, two flagellar proteins FlhA and FlgI. These scFvs have been used in ELISA and dot blot assays against purified protein antigens and 'Ca. Liberibacter asiaticus' infected plant extracts. We have also recloned many of these scFvs into a plasmid expression vector designed for the production of scFvs. Screening of these scFvs was more efficient when phage-bound, rather than soluble scFvs, were used. We have demonstrated a technology to produce antibodies at will and against any protein target encoded by 'Ca. Liberibacter asiaticus'. Applications could include advanced diagnostic methods for huanglongbing and the development of immune labeling reagents for in planta applications. PMID:26744234

  20. Radiolabeled monoclonal antibodies. Development of a new method to remove circulating activity - diagnostic applications and implications for therapy

    International Nuclear Information System (INIS)

    The aim of this thesis was to develop and investigate the usefulness of extracorporeal immunoadsorption (ECIA) to remove circulating activity after the localization of radiolabeled monoclonal antibodies to tumors. A compartment model, based on the biokinetics of 125I-labeled antibodies 96.5 was developed to estimate the effect of ECIA on the tumor-to-normal tissue ratios. ECIA was simulated at different times after injection of the antibodies and the calculations showed an increased diagnostic ratio for several hours after the ECIA procedure, and that an enhancement of the therapeutic ratio was possible. These results led to the development of animal models where the ECIA could be evaluated and the biokinetic behaviour of different radiolabeled monoclonal antibodies investigated with and without the application of ECIA. A general ECIA method, based on biotinylated antibodies and an avidin agarose column as adsorbent, was developed. Studies in tumor bearing nude rats showed that ECIA enhanced of the tumor-to-normal tissue activity ratios by a factor of 4 for the liver, kidneys and bone marrow. For the L6 antibody, the image contrast of tumors localized in one kidney of the rats, was increased from 1.1 to 1.6. A software anthropomorphic phantom was used in Monte Carlo simulations of clinically realistic scintillation camera image acquisition. The effect of ECIA on the contrast enhancement and on the detectability of simulated tumors located centrally in the liver was studied. The contrast increased linearly with an increasing tumor/liver ratio. The contrast was higher for SPECT than for planar images and a contrast of 1.15 required a tumor/liver activity ratio of 1.9 for SPECT and 4.5 for planar images. ECIA in combination with SPECT imaging of radiolabeled antibodies has a great potential in increasing the detectability of tumors. These studies have shown the possibility with ECIA to increase the contrast in radioimmunoimaging and to enhance the therapeutic ratio

  1. Development for advanced materials and testing techniques

    Energy Technology Data Exchange (ETDEWEB)

    Hishinuma, Akimichi [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    1998-03-01

    Recent studies using a JMTR and research reactors of JRR-2 and JRR-3 are briefly summarized. Small specimen testing techniques (SSTT) required for an effective use of irradiation volume and also irradiated specimens have been developed focussing on tensile test, fatigue test, Charpy test and small punch test. By using the small specimens of 0.1 - several mm in size, similar values of tensile and fatigue properties to those by standard size specimens can be taken, although the ductile-brittle transition temperature (DBTT) depends strongly on Charpy specimen size. As for advanced material development, R and D about low activation ferritic steels have been done to investigate irradiation response. The low activation ferritic steel, so-called F82H jointly-developed by JAERI and NKK for fusion, has been confirmed to have good irradiation resistance within a limited dose and now selected as a standard material in the fusion material community. It is also found that TiAi intermetallic compounds, which never been considered for nuclear application in the past, have an excellent irradiation resistance under an irradiation condition. Such knowledge can bring about a large expectation for developing advanced nuclear materials. (author)

  2. The Correlation between Sexual Practices and the Development of Antisperm Antibodies

    Directory of Open Access Journals (Sweden)

    Reza Salman Yazdi

    2009-01-01

    Full Text Available Background: Infertility is one of the most common and important subjects in today’s obstetricsand gynecology. Immunological factors such as the presence of antisperm antibodies (ASA arechallenging etiologies for infertility. This study was performed to determine the correlation betweenthe type of sexual practices (oral‚ anal and vaginal during menstruation and the ASA levels insemen and in the sexual partners’ serum.Materials and Methods: In this analytic cross sectional study which was performed in RoyanInstitute between 2005-2007‚ the type of sexual behaviours was determined in 51 couples withprimary or secondary infertility. The ASA level was determined in both sexual partners’ bloodserum and in the semen‚ using the Sperm Mar Test kit.Results: Using statistical analyses‚ there was no significant difference between the types of sexualpractices (anal‚ oral‚ vaginal during menstruation and the prevalence and level of ASA.Conclusion: Based on the results of this study, the prevalence and level of ASA has no significantcorrelation with the types of sexual behaviours (anal‚ oral‚ vaginal during menstruation.

  3. Development of a multiplex microsphere immunoassay for the quantitation of salivary antibody responses to selected waterborne pathogens

    Science.gov (United States)

    Saliva has an important advantage over serum as a medium for antibody detection due to non-invasive sampling, which is critical for community-based epidemiological surveys. The development of a Luminex multiplex immunoassay for measurement of salivary IgG and IgA responses to pot...

  4. Development of an automated ultrasonic testing system

    Science.gov (United States)

    Shuxiang, Jiao; Wong, Brian Stephen

    2005-04-01

    Non-Destructive Testing is necessary in areas where defects in structures emerge over time due to wear and tear and structural integrity is necessary to maintain its usability. However, manual testing results in many limitations: high training cost, long training procedure, and worse, the inconsistent test results. A prime objective of this project is to develop an automatic Non-Destructive testing system for a shaft of the wheel axle of a railway carriage. Various methods, such as the neural network, pattern recognition methods and knowledge-based system are used for the artificial intelligence problem. In this paper, a statistical pattern recognition approach, Classification Tree is applied. Before feature selection, a thorough study on the ultrasonic signals produced was carried out. Based on the analysis of the ultrasonic signals, three signal processing methods were developed to enhance the ultrasonic signals: Cross-Correlation, Zero-Phase filter and Averaging. The target of this step is to reduce the noise and make the signal character more distinguishable. Four features: 1. The Auto Regressive Model Coefficients. 2. Standard Deviation. 3. Pearson Correlation 4. Dispersion Uniformity Degree are selected. And then a Classification Tree is created and applied to recognize the peak positions and amplitudes. Searching local maximum is carried out before feature computing. This procedure reduces much computation time in the real-time testing. Based on this algorithm, a software package called SOFRA was developed to recognize the peaks, calibrate automatically and test a simulated shaft automatically. The automatic calibration procedure and the automatic shaft testing procedure are developed.

  5. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    OpenAIRE

    Anju Mohan; Hari Mohan Saxena; Puneet Malhotra

    2016-01-01

    Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT), microtiter plate agglutination test (MAT), indirect hemagglutination assay (IHA), and indirect enzyme-linked immunosorbent assay (iELISA) as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cat...

  6. CICC Joint Development and Test for the Test Facility

    Institute of Scientific and Technical Information of China (English)

    武玉; 翁佩德

    2005-01-01

    The superconducting joint of the NbTi Cable-in -conduit Conductor (CICC) has been developed and tested on the magnet test facility at Institute of Plasma Physics, Chinese Academy of Sciences. The CICC is composed of (2NbTi+1Cu)×3×3×(6+1tube) strands each with 0.85 mm in diameter, which has been developed for a central solenoid model coil. The effective length of the joint is about 500 mm. There have been two common fabrication modes,one of them is to integrate the 2 CICC terminals with the copper substrate via lead-soldering, and the other is to mechanically compress the above two parts into an integrated unit. In the current range from 2 kA to 10 kA the joint resistance changes slightly. Up to now, 11 TF magnets, a central solenoid model coil, a central solenoid prototype coil, and a large PF model coil of PF large coil have been completed via the latter joint in the test facility.

  7. Ice slurry cooling development and field testing

    Energy Technology Data Exchange (ETDEWEB)

    Kasza, K.E. [Argonne National Lab., IL (United States); Hietala, J. [Northern States Power Co., Minneapolis, MN (United States); Wendland, R.D. [Electric Power Research Inst., Palo Alto, CA (United States); Collins, F. [USDOE, Washington, DC (United States)

    1992-07-01

    A new advanced cooling technology collaborative program is underway involving Argonne National Laboratory (ANL), Northern States Power (NSP) and the Electric Power Research Institute (EPRI). The program will conduct field tests of an ice slurry distributed load network cooling concept at a Northern States Power utility service center to further develop and prove the technology and to facilitate technology transfer to the private sector. The program will further develop at Argonne National Laboratory through laboratory research key components of hardware needed in the field testing and develop an engineering data base needed to support the implementation of the technology. This program will sharply focus and culminate research and development funded by both the US Department of Energy and the Electric Power Research Institute on advanced cooling and load management technology over the last several years.

  8. Bell Canyon Test (BCT): cement development report

    International Nuclear Information System (INIS)

    The Borehole Plugging (BHP) materials development program which has been underway at WES under Sandia sponsorship for about five years is reviewed. Development testing data for candidate grout mixtures for the BCT plug are presented. Field batching, mixing, and placement operations are discussed. Data from field samples molded during the two plug placements include strength, expansion, compressional wave velocity, dynamic modulus, density, and porosity. Microstructure and composition are compared for grout samples at ages of a few weeks and one year

  9. Research and Development Trends of Antibody Therapeutics%治疗性抗体药物研究与发展趋势

    Institute of Scientific and Technical Information of China (English)

    王旻

    2011-01-01

    The advent of monoclonal antibody technology actualized the research and manufacture of monoclonal antibodies. With the development of gene engineering, recombinant genetic antibody came true. With the aid of recombinant DNA technology, murine antibody could be humanized, and some synthetic or semisynthetic human antibody library or phage displayed human antibody library have been constructed. Therefore, human antibody could be obtained by screening from such libraries. Transgenic mouse is another source of human antibody. The class of therapeutic antibodies are varied from munne antibody to human - mouse chimeric antibody,1 to humanized antibody and finally human antibody. Most of the recently approved therapeutic antibodies are derived from human antibody. From the year of 1996 to 2008, 45 percent of the monoclonal antibodies in clinical trial are anti tumor antibodies, and 28 percent are for immunologic derangement. In conclusion the development of therapeutic antibodies has run into a virtuous cycle from R&D to return, which lead to a hotspot in the international pharmaceutical industries. In this article, the history, market, problem and perspective of therapeutic antibodies are highlighted.%单克隆抗体技术的问世,使研究和生产治疗性单抗药物成为现实.随着基因工程技术的发展,新型的重组抗体技术也随之而生.人们可以利用DNA重组技术对鼠源抗体进行人源化改造、构建合成或半合成抗体库及噬菌体抗体库,从中筛选获得人源抗体,甚至利用转基因小鼠直接获得人源抗体.抗体药物发展的趋势也从鼠源、人一鼠嵌合、人源化到全人源.近年获得批准的抗体药物以全人源为主.1996年至2008年间进入临床研究的人源化单克隆抗体中45%用于治疗肿瘤,28%个用于治疗免疫紊乱.抗体药物的发展进入研发、回报的良性循环,成了国际制药业争夺的焦点.文章就治疗性抗体发展的历史、现状、市场及未来展望作了简要综述.

  10. Seroprevalence of Neospora caninum and Toxoplasma gondii antibodies in white tailed deer (odocoileus virginianus) from Iowa and Minnesota using four serologic tests

    Science.gov (United States)

    The white-tailed deer (Odocoileus virginianus) is considered one of the most important wildlife reservoir of Neospora caninum and Toxoplasma gondii in the US. Sera from white-tailed deer from Minnesota and Iowa were tested for antibodies to N. caninum by four serologic tests including the indi...

  11. Serum Neutralization Assay Can Efficiently Replace Plaque Reduction Neutralization Test for Detection and Quantitation of West Nile Virus Antibodies in Human and Animal Serum Samples

    Science.gov (United States)

    Di Gennaro, Annapia; Casaccia, Claudia; Conte, Annamaria; Monaco, Federica; Savini, Giovanni

    2014-01-01

    A serum neutralization assay (SN) was compared with the official plaque reduction neutralization test for the quantitation of West Nile virus antibodies. A total of 1,348 samples from equid sera and 38 from human sera were tested by these two methods. Statistically significant differences were not observed, thus supporting the use of SN for routine purposes. PMID:25100824

  12. SRS environmental technology development field test platform

    International Nuclear Information System (INIS)

    A critical and difficult step in the development and implementation of new technologies for environmental monitoring and characterization is successfully transferring these technologies to industry and government users for routine assessment and compliance activities. The Environmental Sciences Section of the DOE Savannah River Technology Center provides a forum for developers, potential users, and regulatory organizations to evaluate new technologies in comparison with baseline technologies in a well characterized field test bed. The principal objective of this project is to conduct comprehensive, objective field tests of monitoring and characterization technologies that are not currently used in EPA standard methods and evaluate their performance during actual operating conditions against baseline methods. This paper provides an overview of the field test site and a description of some of the technologies demonstrated at the site including their field applications

  13. Detection of anti-U3-RNP/fibrillarin IgG antibodies by line immunoblot assay has comparable clinical significance to immunoprecipitation testing in systemic sclerosis.

    Science.gov (United States)

    Peterson, Lisa K; Jaskowski, Troy D; Mayes, Maureen D; Tebo, Anne E

    2016-04-01

    The aim of this study was to evaluate the performance and clinical relevance of a commercially available line immunoblot assay (LIA) for detecting anti-U3-RNP/fibrillarin (anti-U3-RNP), against immunoprecipitation (gold standard). This study involved a multi-ethnic cohort of 1000 American systemic sclerosis (SSc) patients and 50 healthy controls. Antinuclear antibodies and centromere antibodies were detected by indirect immunofluorescent antibody test, anti-topo I by immunodiffusion and anti-RNAP III by ELISA. The presence of anti-U3-RNP in select serum samples was detected by immunoprecipitation (IP) and LIA. By IP, U3-RNP antibody was detected in 75 (7.5 %) patients with SSc. Overall agreement between LIA and IP was very good (κ = 0.966). Analytic sensitivity and specificity of the U3-RNP LIA was 100 and 94.7 %, respectively. Clinical features associated with positivity for the anti-U3-RNP antibody include diffuse cutaneous SSc and increased prevalence of renal crisis, consistent with previous studies that used IP. Testing for U3-RNP antibodies is only performed by a small number of laboratories due to the complexity of both performance and interpretation of the IP. LIA is faster and less complex than IP. Excellent agreement between IP and LIA demonstrates that LIA is an acceptable and attractive alternative to IP for anti-U3-RNP detection. PMID:26467972

  14. Next Generation Drivetrain Development and Test Program

    Energy Technology Data Exchange (ETDEWEB)

    Keller, Jonathan; Erdman, Bill; Blodgett, Doug; Halse, Chris; Grider, Dave

    2015-11-03

    This presentation was given at the Wind Energy IQ conference in Bremen, Germany, November 30 through December 2, 2105. It focused on the next-generation drivetrain architecture and drivetrain technology development and testing (including gearbox and inverter software and medium-voltage inverter modules.

  15. Developing a solar panel testing system

    Directory of Open Access Journals (Sweden)

    Árpád Rácz

    2009-10-01

    Full Text Available Solar energy is increasingly used togenerate electricity for individual households. There isa wide variety of solar panel technologies, whichshould be tested at an individual level during theirlifetime. In this paper, the development of a testingstation at the University of Debrecen is presented. Thetesting system can be used for research andeducational purposes and for in field applicationsequally well.

  16. Interactive Test Analysis: Development, Implementation, and Evaluation.

    Science.gov (United States)

    Lipe, Gary

    An interactive test analysis system was developed which interfaces a 3M DATRONICS system with a XEROX Sigma 9 computer. The computer programs were written in A Programming Language (APL). The current implementation of the program is characterized by its capability to: read responses from a DATRONIC answer sheet; allow the faculty member the option…

  17. Evaluation of an egg yolk enzyme-linked immunosorbent assay antibody test and its use to assess the prevalence of Mycoplasma gallisepticum infection in laying hens in Italy

    Directory of Open Access Journals (Sweden)

    Marco Tamba

    2010-01-01

    Full Text Available The prevalence of Mycoplasma gallisepticum infection in commercial layers was established by the presence of antibodies in eggs. Saline-extracted yolks were used with a commercial enzyme-linked immunosorbent assay kit. For the prevalence study, yolks from 30 eggs were obtained from each of 66 flocks coming from 36 layer farms. The prevalence of egg antibodies to Mycoplasma gallisepticum was 33.3% in single-age farms and 77.8% in multi-age farms. In 27 flocks, antibody titers were compared with results obtained from blood samples taken in the same flock and in the same period and analyzed with the same kit. This study has confirmed that egg yolk enzyme-linked immunosorbent assay antibody test is a suitable and practical approach for assessing the flock prevalence of Mycoplasma gallisepticum infection in layer hens.

  18. Antissaliva Antibodies of Lutzomyia Longipalpis in area of Visceral Leishmaniasis.

    Science.gov (United States)

    Fraga, Thiago Leite; Fernandes, Magda Freitas; Pontes, Elenir Rose Jardim Cury; Levay, Ana Paula Silva; Almeida da Cunha, Elenice Brandão; França, Adriana de Oliveira; Dorval, Maria Elizabeth Cavalheiros

    2016-07-01

    The aim of the present study was to assess the presence of antissaliva antibodies of Lutzomyia longipalpis in human hosts living in area of visceral leishmaniasis, located in the Center-West region of Brazil. The presence of antissaliva antibodies of L. longipalpis exhibited a strong correlation with the protection and development of antibodies against Leishmania sp. Of the 492 children studied, elevated antissaliva antibodies of L. longipalpis were detected in 38.4% of the participants. There was a higher percentage of positivity (64.7%) among children who exhibited anti-Leishmania sp. antibodies and among those who were positive in the delayed hypersensitivity test (34.8%). PMID:27093167

  19. The antibody mining toolbox

    OpenAIRE

    D'Angelo, Sara; Glanville, Jacob; Ferrara, Fortunato; Naranjo, Leslie; Gleasner, Cheryl D.; Shen, Xiaohong; Bradbury, Andrew RM; Kiss, Csaba

    2013-01-01

    In vitro selection has been an essential tool in the development of recombinant antibodies against various antigen targets. Deep sequencing has recently been gaining ground as an alternative and valuable method to analyze such antibody selections. The analysis provides a novel and extremely detailed view of selected antibody populations, and allows the identification of specific antibodies using only sequencing data, potentially eliminating the need for expensive and laborious low-throughput ...

  20. Quality and quantity of TFH cells are critical for broad antibody development in SHIVAD8 infection.

    Science.gov (United States)

    Yamamoto, Takuya; Lynch, Rebecca M; Gautam, Rajeev; Matus-Nicodemos, Rodrigo; Schmidt, Stephen D; Boswell, Kristin L; Darko, Sam; Wong, Patrick; Sheng, Zizhang; Petrovas, Constantinos; McDermott, Adrian B; Seder, Robert A; Keele, Brandon F; Shapiro, Lawrence; Douek, Daniel C; Nishimura, Yoshiaki; Mascola, John R; Martin, Malcolm A; Koup, Richard A

    2015-07-29

    Broadly neutralizing antibodies (bNAbs) protect against HIV-1 infection, yet how they are generated during chronic infection remains unclear. It is known that T follicular helper (TFH) cells are needed to promote affinity maturation of B cells during an immune response; however, the role of TFH during HIV-1 infection is undefined within lymph node germinal centers (GCs). We use nonhuman primates to investigate the relationship in the early stage of chronic SHIVAD8 (simian-human immunodeficiency virus AD8) infection between envelope (Env)-specific TFH cells, Env-specific B cells, virus, and the generation of bNAbs during later infection. We found that both the frequency and quality of Env-specific TFH cells were associated with an expansion of Env-specific immunoglobulin G-positive GC B cells and broader neutralization across HIV clades. We also found a correlation between breadth of neutralization and the degree of somatic hypermutation in Env-specific memory B cells. Finally, we observed high viral loads and greater diversity of Env sequences in rhesus macaques that developed cross-reactive neutralization as compared to those that did not. These studies highlight the importance of boosting high-quality TFH populations as part of a robust vaccine regimen aimed at eliciting bNabs. PMID:26223303

  1. Development of radioimmunotherapeutic and diagnostic antibodies: an inside-out view

    Energy Technology Data Exchange (ETDEWEB)

    Boswell, C. Andrew [Radioimmune and Inorganic Chemistry Section, Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1088 (United States); Brechbiel, Martin W. [Radioimmune and Inorganic Chemistry Section, Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1088 (United States)], E-mail: martinwb@mail.nih.gov

    2007-10-15

    Only a handful of radiolabeled antibodies (Abs) have gained US Food and Drug Administration (FDA) approval for use in clinical oncology, including four immunodiagnostic agents and two targeted radioimmunotherapeutic agents. Despite the advent of nonimmunogenic Abs and the availability of a diverse library of radionuclides, progress beyond early Phase II radioimmunotherapy (RIT) studies in solid tumors has been marginal. Furthermore, [{sup 18}F]fluorodeoxyglucose continues to dominate the molecular imaging domain, underscored by a decade-long absence of any newly approved Ab-based imaging agent (none since 1996). Why has the development of clinically successful Abs for RIT been limited to lymphoma? What obstacles must be overcome to allow the FDA approval of immuno-positron emission tomography (immuno-PET) imaging agents? How can we address the unique challenges that have thus far prevented the introduction of Ab-based imaging agents and therapeutics for solid tumors? Many poor decisions have been made regarding radiolabeled Abs, but useful insight can be gained from these mistakes. The following review addresses the physical, chemical, biological, clinical, regulatory and financial limitations that impede the progress of this increasingly important class of drugs.

  2. Quality Control System for Beer Developed with Monoclonal Antibodies Specific to Barley Lipid Transfer Protein

    Directory of Open Access Journals (Sweden)

    Yukie Murakami-Yamaguchi

    2012-10-01

    Full Text Available Non-specific lipid transfer protein (LTP in barley grain reacted with the IgE in sera drawn from food allergy patients. A sandwich-type of enzyme-linked immunosorbent assay (ELISA was developed with mouse monoclonal antibodies raised against LTP purified with barley flour. This ELISA showed a practical working range of 0.3–3 ng/mL and no cross-reactivity with wheat, adlay and rye. Using this ELISA, LTP was determined in several types of barley-foods, including fermented foods such as malt vinegar, barley-malt miso and beer. LTP content in beer of the same kind was approximately constant, even if manufacturing factory and production days were different. Not only as a factor of foam formation and stability but also as an allergen, controlling and monitoring of LTP in beer should be considered. Taken together, our LTP-detecting ELISA can be proposed as an appropriate system for the quality control of beer.

  3. Development of antibody-siRNA conjugate targeted to cardiac and skeletal muscles.

    Science.gov (United States)

    Sugo, Tsukasa; Terada, Michiko; Oikawa, Tatsuo; Miyata, Kenichi; Nishimura, Satoshi; Kenjo, Eriya; Ogasawara-Shimizu, Mari; Makita, Yukimasa; Imaichi, Sachiko; Murata, Shumpei; Otake, Kentaro; Kikuchi, Kuniko; Teratani, Mika; Masuda, Yasushi; Kamei, Takayuki; Takagahara, Shuichi; Ikeda, Shota; Ohtaki, Tetsuya; Matsumoto, Hirokazu

    2016-09-10

    Despite considerable efforts to develop efficient carriers, the major target organ of short-interfering RNAs (siRNAs) remains limited to the liver. Expanding the application outside the liver is required to increase the value of siRNAs. Here we report on a novel platform targeted to muscular organs by conjugation of siRNAs with anti-CD71 Fab' fragment. This conjugate showed durable gene-silencing in the heart and skeletal muscle for one month after intravenous administration in normal mice. In particular, 1μg siRNA conjugate showed significant gene-silencing in the gastrocnemius when injected intramuscularly. In a mouse model of peripheral artery disease, the treatment with myostatin-targeting siRNA conjugate by intramuscular injection resulted in significant silencing of myostatin and hypertrophy of the gastrocnemius, which was translated into the recovery of running performance. These data demonstrate the utility of antibody conjugation for siRNA delivery and the therapeutic potential for muscular diseases. PMID:27369865

  4. Transfusion management of patients with red blood cell antibodies

    Directory of Open Access Journals (Sweden)

    Bujandrić Nevenka B.

    2013-01-01

    Full Text Available Introduction. Red blood cell antibodies may cause a positive result of pre-transfusion blood compatibility testing (crossmatch test. It can be a problem to provide suitable blood units for patients with clinically significant antibodies to high-frequency antigens as well as for those with multiple alloantibody specificities. This study was aimed at identifying transfused patients in the population of South-Backa who had developed clinically significant red blood cell alloantibodies. Material and methods. We analyzed the records of crossmatch results and antibody screening performed at the Blood Transfusion Institute of Vojvodina during 2012. Results. Antibodies were found in 103 patients: A 63 patients with single antibodies: 1 16 with antibodies of unknown specificity (3 autoantibodies, 13 alloantibodies; 2 39 with clinically significant antibodies (23 from Rh system (2 anti-C, 2 anti-D, 12 anti-E, 7 anti-c, 4 anti-K, 3 anti-Fya, 7 anti-Jka, 2 anti-S; 3 8 with usually not significant antibodies (6 anti-M, 1 anti-A1, 1 anti- Cw; B 40 patients developed multiple antibodies: 1 all patients had at least one clinically significant antibody from various blood group system (44 Rh, 13 Kell, 7 Kidd, 7 MNSs (S, s; 2 3 patients had usually not significant antibodies (1 Lewis, 2 Lutheran; 3 3 patients occasionally had clinically significant antibody (3 anti- Yta; 4 3 patients had antibodies of unknown specificity (2 autoantibodies, 1alloantibody. Antibodies detected in the majority of patients (65-63.1% had a specificity of Rh and/or the Kell system. Conclusions. The main goal of pre-transfusion blood compatibility testing is to detect clinically significant antibodies. The provision of antigen negative blood units for those patients is a special challenge for blood establishments. Database with a sufficient number of typed blood donors can help to resolve this problem.

  5. Detection of rainbow trout antibodies against viral haemorrhagic septicaemia virus (VHSV) by neutralisation test is highly dependent on the virus isolate used

    DEFF Research Database (Denmark)

    Fregeneda-Grandes, J.M.; Olesen, Niels Jørgen

    2007-01-01

    50 %PNT, 90 % of the fish were found to be positive. By examining a panel of different VHSV isolates in 50 %PNT, it was demonstrated that the virus isolate used as test antigen could significantly affect the sensitivity and titre determination in 50 %PNT for detection of rainbow trout antibodies......Three serological tests, enzyme linked immunosorbent assay (ELISA), 50 % plaque neutralisation test (50%PNT) and Western blotting (WB), were used to detect antibodies against viral haemorrhagic septicaerma virus (VHSV) in 50 rainbow trout broodstock from a rainbow trout farm endemically infected...... against VHSV. When the sera were examined for the presence of VHSV antibodies by ELISA or WB, 61 % were found to be positive. When conducting WB analysis, the viral glycoprotein was the protein most frequently recognized, followed by the viral nucleoprotein....

  6. Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.

    Science.gov (United States)

    Fernandes, Cláudia P H; Seixas, Fabiana K; Coutinho, Mariana L; Vasconcellos, Flávia A; Seyffert, Núbia; Croda, Julio; McBride, Alan J; Ko, Albert I; Dellagostin, Odir A; Aleixo, José A G

    2007-02-01

    Pathogenic serovars of Leptospira have a wide antigenic diversity attributed mainly to the lipopolysaccharide present in the outer membrane. In contrast, antigens conserved among pathogenic serovars are mainly represented by outer membrane proteins. Surface exposure of a major and highly conserved outer membrane lipoprotein (LipL32) was recently demonstrated on pathogenic Leptospira. LipL32 in its recombinant form (rLipL32) was used to immunize BALB/c mice to develop murine monoclonal antibodies (MAbs). Three MAbs against rLipL32 were produced, isotyped, and evaluated for further use in diagnostic tests of leptospirosis using different approaches. MAbs were conjugated to peroxidase and evaluated in a native protein enzyme-linked immunosorbent assay (ELISA) with intact and heat-treated leptospiral cells, conjugated to fluorescein isothiocyanate (FITC) for indirect immunofluorescence with intact and methanol fixed cells and were used for LipL32 immunoprecipitation from leptospiral cells. rLipL32 MAbs conjugated to peroxidase or used as primary antibody bound to intact and heat-treated cells in ELISA, proving that they could be used in enzyme immunoassays for detection of the native protein. In immunofluorescence assay, MAbs labeled bacterial cells either intact or methanol fixed. Two MAbs were able to immunoprecipitate the native protein from live and motile leptospiral cells and, adsorbed onto magnetic beads, captured intact bacteria from artificially contaminated human sera for detection by polymerase chain reaction (PCR) amplification. Results of this study suggest that the MAbs produced can be useful for the development of diagnostic tests based on detection of LipL32 leptospiral antigen in biological fluids. PMID:17316084

  7. A Peptide-Based Plasmodium falciparum Circumsporozoite Assay To Test for Serum Antibody Responses to Pre-Erythrocyte Malaria Vaccines ▿

    OpenAIRE

    Kostense, Stefan; Mommaas, Bregje; Hendriks, Jenny; Verhoeven, Mariëlle; ter Haak, Mariska; Tirion, Felicia; Wiesken, Edison; Pau, Maria Grazia; Radošević, Katarina; Goudsmit, Jaap

    2011-01-01

    Various pre-erythrocyte malaria vaccines are currently in clinical development, and among these is the adenovirus serotype 35-based circumsporozoite (CS) vaccine produced on PER.C6 cells. Although the immunological correlate of protection against malaria remains to be established, the CS antibody titer is a good marker for evaluation of candidate vaccines. Here we describe the validation of an anti-Plasmodium falciparum circumsporozoite antibody enzyme-linked immunosorbent assay (ELISA) based...

  8. Longitudinal Monitoring of the Development of Antifilarial Antibodies and Acquisition of Wuchereria bancrofti in a Highly Endemic Area of Haiti

    OpenAIRE

    Joseph Kubofcik; Fink, Doran L.; Nutman, Thomas B.

    2012-01-01

    BACKGROUND: The current antibody tests used for monitoring in lymphatic filariasis (LF) elimination programs suffer from poor specificity because of the considerable geographical overlap with other filarial infections such as Loa loa (Ll), Onchocerca volvulus (Ov), and Mansonella perstans (Mp). METHODS: Using bioinformatics to assemble into contigs 2048 expressed sequence tags (ESTs) from the L3 infective larvae of W. bancrofti (Wb), these were next assessed for homology to known proteins and...

  9. Development and validation of an enzyme-linked immunosorbent assay for antibodies against Mycobacterium bovis in european wild boar

    Directory of Open Access Journals (Sweden)

    Gortázar Christian

    2008-11-01

    Full Text Available Abstract Background Bovine tuberculosis (bTB remains a significant problem in some parts of Spain largely because of contacts between cattle and wildlife reservoirs in extensive grazing systems. European Wild boar (Sus scrofa is one of the species involved in the transmission of the disease to other species. Fast and simple detection methods would be critical for assessing infection prevalence, study the mechanisms of pathogen transmission and monitoring the effects of TB control measures. Results An enzyme-linked immunosorbent assay (ELISA to detect antibodies against Mycobacterium bovis in wild boar serum was developed and validated on 185 sera from TB positive and negative wild boar. Based on antigen inoculation of captive animals as well as tuberculosis compatible lesions, culture results and molecular analysis of hunted individuals, animals were allocated into two groups: tuberculosis positive group and tuberculosis negative group. After optimization of the positive to negative ratio using different combinations of serum dilutions and conjugate concentrations, the test yielded a sensitivity of 72.60% and a specificity of 96.43% for the best cut-off. Conclusion Although some negative group animals showed an ELISA positive reaction (

  10. Avian Diagnostic and Therapeutic Antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Bradley, David Sherman [UND SMHS

    2012-12-31

    A number of infectious agents have the potential of causing significant clinical symptomology and even death, but dispite this, the number of incidence remain below the level that supports producing a vaccine. Therapeutic antibodies provide a viable treatment option for many of these diseases. We proposed that antibodies derived from West Nile Virus (WNV) immunized geese would be able to treat WNV infection in mammals and potential humans. We demonstrated that WNV specific goose antibodies are indeed successful in treating WNV infection both prophylactically and therapeutically in a golden hamster model. We demonstrated that the goose derived antibodies are non-reactogenic, i.e. do not cause an inflammatory response with multiple exposures in mammals. We also developed both a specific pathogen free facility to house the geese during the antibody production phase and a patent-pending purification process to purify the antibodies to greater than 99% purity. Therefore, the success of these study will allow a cost effective rapidly producible therapeutic toward clinical testing with the necessary infrastructure and processes developed and in place.

  11. Development and testing of a mobile incinerator

    International Nuclear Information System (INIS)

    The development and testing of a mobile incinerator for processing of combustible dry active waste (DAW) and contaminated oil generated at Nuclear Power Plants is presented. Topics of discussion include initial thoughts on incineration as applied to nuclear waste; DOE's Aerojet's, and CECo's role in the Project; design engineering concepts; site engineering support; licensability; generation of test data; required reports of the NRC and Illinois and California EPA's; present project schedule for incinerating DAW at Dresden and other CECo Stations; and lessons learned from the project

  12. [Challenge to the Development of Molecular Targeted Therapy against a Novel Target Candidate Identified by Antibody Proteomics Technology].

    Science.gov (United States)

    Nagano, Kazuya

    2016-01-01

    Disease proteomics that systemically analyzes and identifies differentially expressed proteins between healthy and diseased samples is a potentially useful approach for obtaining target proteins for drug development. To date, however, very few target proteins have been identified from this field. A key issue that remains to be resolved is how to correctly identify target proteins from a number of potential candidates. To circumvent this problem, we have developed "antibody proteomics technology" in which a single chain Fv phage antibody library is utilized for proteome analysis. Here, we describe the application of this technology by primarily focusing on Eph receptor A10 (EphA10), a novel breast cancer-related protein that is a promising target for antibody drugs. To establish an effective and safe targeted cancer therapy, it is important that the target is specifically expressed in cancer tissues. Therefore, we attempted to analyze the EphA10 expression profiles. Tissue microarray analysis showed that EphA10 was expressed in all subtypes of breast cancer containing triple negative breast cancer cases. On the other hand, EphA10 was only expressed in testis tissue among 36 kinds of normal tissues. Thus, EphA10 could be a highly cancer-specific protein, making it a promising target for female breast cancer patients. Finally, we examined the anti-tumor effect by anti-EphA10 antibody, aiming for the development of a novel EphA10 targeting therapy. Administration of the antibody showed that tumor volumes were significantly inhibited. Our results suggest that targeting EphA10 in breast cancer cases might be a promising new form of therapy. PMID:26831784

  13. Hapten synthesis and antibody production for the development of a melamine immunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Lei Hongtao; Shen Yudong; Song Lijun; Yang Jinyi [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Chevallier, Olivier P.; Haughey, Simon A. [Institute of Agri-Food and Land Use, Queen' s University Belfast, Belfast BT9 5AG, Northern Ireland (United Kingdom); Wang Hong [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Sun Yuanming, E-mail: ymsun@scau.edu.cn [Key Laboratory of Food Safety of Guangdong Province/Institute of Food Quality and Safety, South China Agricultural University, Wushan Road, Tianhe District, Guangzhou 510642, Guangdong (China); Elliott, Christopher T., E-mail: chris.elliott@qub.ac.uk [Institute of Agri-Food and Land Use, Queen' s University Belfast, Belfast BT9 5AG, Northern Ireland (United Kingdom)

    2010-04-14

    The incorporation of melamine into food products is banned but its misuse has been widely reported in both animal feeds and food. The development of a rapid screening immunoassay for monitoring of the substance is an urgent requirement. Two haptens of melamine were synthesized by introducing spacer arms of different lengths and structures on the triazine ring of the analyte molecular structure. 6-Aminocaproic acid and 3-mercaptopropionic acid were reacted with 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) to produce hapten 1 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylamino) hexanoic acid] and hapten 2 [3-(4,6-diamino-1,6-dihydro-1,3,5-triazin-2-ylthio) propanoic acid], respectively. The molecular structures of the two haptens were identified by {sup 1}H nuclear magnetic resonance spectrometry, mass spectrometry and infrared spectrometry. An immunogen was prepared by coupling hapten 1 to bovine serum albumin (BSA). Two plate coating antigens were prepared by coupling both haptens to egg ovalbumin (OVA). A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was developed to evaluate homogeneous and heterogeneous assay formats. The results showed that polyclonal antibodies with high titers were obtained, and the heterogeneous immunoassay format demonstrated a better performance with an IC{sub 50} of 70.6 ng mL{sup -1}, a LOD of 2.6 ng mL{sup -1} and a LOQ of 7.6 ng mL{sup -1}. Except for cyromazine, no obvious cross-reactivity to common compounds was found. The data showed that the hapten synthesis was successful and the resultant antisera could be used in an immunoassay for the rapid and sensitive detection of this banned chemical.

  14. Protection of recombinant mammalian antibodies from development-dependent proteolysis in leaves of Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Stéphanie Robert

    Full Text Available The expression of clinically useful proteins in plants has been bolstered by the development of high-yielding systems for transient protein expression using agroinfiltration. There is a need now to know more about how host plant development and metabolism influence the quantity and quality of recombinant proteins. Endogenous proteolysis is a key determinant of the stability and yield of recombinant proteins in plants. Here we characterised cysteine (C1A and aspartate (A1 protease profiles in leaves of the widely used expression host Nicotiana benthamiana, in relation with the production of a murine IgG, C5-1, targeted to the cell secretory pathway. Agroinfiltration significantly altered the distribution of C1A and A1 proteases along the leaf age gradient, with a correlation between leaf age and the level of proteolysis in whole-cell and apoplast protein extracts. The co-expression of tomato cystatin SlCYS8, an inhibitor of C1A proteases, alongside C5-1 increased antibody yield by nearly 40% after the usual 6-days incubation period, up to ~3 mg per plant. No positive effect of SlCYS8 was observed in oldest leaves, in line with an increased level of C1A protease activity and a very low expression rate of the inhibitor. By contrast, C5-1 yield was greater by an additional 40% following 8- to 10-days incubations in younger leaves, where high SlCYS8 expression was maintained. These findings confirm that the co-expression of recombinant protease inhibitors is a promising strategy for increasing recombinant protein yields in plants, but that further opportunity exists to improve this approach by addressing the influence of leaf age and proteases of other classes.

  15. Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme

    Directory of Open Access Journals (Sweden)

    Dickson Dennis W

    2009-10-01

    Full Text Available Abstract Background Insulin-degrading enzyme (IDE is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs targeting natively folded human and rodent IDE. Results Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts. Conclusion We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.

  16. Development of rabbit monoclonal antibodies for detection of alpha-dystroglycan in normal and dystrophic tissue.

    Directory of Open Access Journals (Sweden)

    Marisa J Fortunato

    Full Text Available Alpha-dystroglycan requires a rare O-mannose glycan modification to form its binding epitope for extracellular matrix proteins such as laminin. This functional glycan is disrupted in a cohort of muscular dystrophies, the secondary dystroglycanopathies, and is abnormal in some metastatic cancers. The most commonly used reagent for detection of alpha-dystroglycan is mouse monoclonal antibody IIH6, but it requires the functional O-mannose structure for recognition. Therefore, the ability to detect alpha-dystroglycan protein in disease states where it lacks the full O-mannose glycan has been limited. To overcome this hurdle, rabbit monoclonal antibodies against the alpha-dystroglycan C-terminus were generated. The new antibodies, named 5-2, 29-5, and 45-3, detect alpha-dystroglycan from mouse, rat and pig skeletal muscle by Western blot and immunofluorescence. In a mouse model of fukutin-deficient dystroglycanopathy, all antibodies detected low molecular weight alpha-dystroglycan in disease samples demonstrating a loss of functional glycosylation. Alternately, in a porcine model of Becker muscular dystrophy, relative abundance of alpha-dystroglycan was decreased, consistent with a reduction in expression of the dystrophin-glycoprotein complex in affected muscle. Therefore, these new rabbit monoclonal antibodies are suitable reagents for alpha-dystroglycan core protein detection and will enhance dystroglycan-related studies.

  17. The testing of antibodies raised against poultry red mite antigens in an in vitro feeding assay; preliminary screen for vaccine candidates.

    Science.gov (United States)

    Wright, Harry W; Bartley, Kathryn; Nisbet, Alasdair J; McDevitt, Regina M; Sparks, Nickolas H C; Brocklehurst, Sarah; Huntley, John F

    2009-06-01

    Dermanyssus gallinae (De Geer), the poultry red mite, is a blood-feeding ectoparasite that infests many bird species. We have used an in vitro feeding assay to allow the identification of protective D. gallinae antigens that may have potential as vaccine candidates. Homogenised mites were extracted sequentially with PBS, Tween 20, Triton X100 and urea giving four protein fractions. Five experimental groups of Lohmann Brown hens were used to generate antibodies; four groups were injected with one of each of the protein fractions in QuilA adjuvant and a control group was injected with adjuvant only. Booster injections were administered 2 and 4 weeks after initial immunisation. Eggs were collected throughout the experiment and soluble IgY antibodies were extracted from a pool of egg yolks collected at week six post-injection. Western blots, performed using post vaccination antibodies from test and control groups, revealed a strong antibody response against a range of injected proteins. Fresh chicken blood, supplemented with antibodies raised against these protein fractions, was fed to mites in an in vitro feeding assay in order to determine whether the antibodies had an anti-mite effect. Although there was variability in the numbers of feeding mites, it was found that the strongest anti-mite effect was seen with the PBS protein fraction, which had a cumulative average mortality of 34.8% 14 days after feeding compared with 27.3% for the control group (P = 0.043). PMID:19184466

  18. Development and Testing of Active Groundwater Samplers

    DEFF Research Database (Denmark)

    Nilsson, Bertel; Jakobsen, Rasmus; Andersen, Lars Jørgen

    1995-01-01

    Active groundwater sampling techniques are methods where the aquifer is flushed by pumping. The methods developed and tested represent non-dedicated methods for use in existing water wells. This paper describes two different sampling techniques: the Separation Pumping Technique (SP) and the Packer...... on numerical modelling and controlled laboratory experiments. Active groundwater sampling techniques can be used for remedial pumping optimization and in obtaining hydraulic data and represent a fast operational and reliable sampling tool, also under heterogeneous and low permeability conditions....

  19. Serologic test systems development. Progress report, July 1, 1976--September 30, 1977

    Energy Technology Data Exchange (ETDEWEB)

    Saunders, G.C.; Clinard, E.H.; Bartlett, M.L.; Petersen, P.M.; Sanders, W.M.; Payne, R.J.; Martinez, E.

    1978-01-01

    Work has continued on the development and application of the Enzyme-Labeled Antibody (ELA) test to the USDA needs. Results on trichinosis, brucellosis, and staphylococcal enterotoxin A detection are very encouraging. A field test for trichinosis detection is being worked out in cooperation with Food Safety and Quality Service personnel. Work is in progress with the Technicon Instrument Corporation to develop a modification of their equipment to automatically process samples by the ELA procedure. An automated ELA readout instrument for 96-well trays has been completed and is being checked out.

  20. Development of a double-antibody radioimmunoassay for the detection of methotrexate in the serum

    International Nuclear Information System (INIS)

    This double-antibody radioimmunoassay (RIA) designed to detect methotrexate (MTX) in the serum is a precise, rapid and cost-saving quantitative procedure, which meets all the requirements regarding accuracy and reproducibility. Its main components are anti-methotrexate as antiserum, 3H-MTX as tracer substance and a second antibody as separating agent. The measuring technique using tritium is based on a method involving removal of the supernate by suction after separation of the antibody, dissolution of the precipitate in NaCl and addition of the scintillator. As opposed to separation techniques based on carbon the radioactivity to be determined is contained in the precipitate, not in the supernate. The procedure permits the required amount of scintillator and the associated radioactive waste to be reduced by 90%. A further decisive advantage is its low limit of detection. (TRV)

  1. Comparative assessment of a double antibody enzyme immunoassay test kit and a triple antibody enzyme immunoassay for the diagnosis of Trichinella spiralis spiralis and Trichinella spiralis nativa infections in swine.

    OpenAIRE

    Smith, H. J.; Snowdon, K E

    1989-01-01

    Enzyme immunoassays using the triple antibody enzyme linked immunosorbent assay (ELISA) with both Trichinella spiralis spiralis and T. spiralis nativa excretory-secretory (ES) antigens and a commercial Trichinella spiralis enzyme immunoassay test kit were carried out on sera from pigs that were infected with light, moderate and high doses of infective T. spiralis spiralis and T. spiralis nativa respectively. Seroconversion occurred in all pigs given infective Trichinella larvae although no tr...

  2. Develope Monoclonal Antibody against Foot-and-mouth Disease Virus A Type

    Institute of Scientific and Technical Information of China (English)

    Tong Lin; Jing Li; Jun-jun Shao; Guo-zheng Cong; Jun-zheng Du; Shan-dian Gao; Hui-yun Chang

    2011-01-01

    In order to develop an anti-FMDV A Type monoclonal antibo by (mAb),BABL/c mice were immunized with FMDV A type.Monoclonal antibodies (mAbs) 7B11 and 8H4 against Foot-and-mouth disease virus (FMDV) serotype A were produced by fusing SP2/O myeloma cells with splenocyte from the mouse immunized with A/AV88.The microneutralization titer of the mAbs 7B11 and 8H4 were 1024 and 512,respectively.Both mAbs contain kappa light chains,the mAbs were IgG1.In order to define the mAbs binding epitopes,the reactivity of these mAbs against A Type FMDV,were examined using indirect ELISA,the result showed that both mAbs reacted with A Type FMDV.These mAbs may be used for further vaccine studies,diagnostic methods,prophylaxis,etiological and immunological research on FMDV.Characterization of these ncindicated that prepared anti-FMDV A mAbs had no cross-reactivity with Swine Vesicular Disease (SVD) or FMDV O,Asial and C Type antigens.Their titers in abdomen liquor were 1:5×106 and 1:2×106,respectively.7B11 was found to be of subtype IgG1,8H4 was classified as IgG2b subtype.The mAbs prepared in this study,are specific for detection of FMDV serotype A,and is potentially useful for pen-side diagnosis.

  3. Progress in the development of therapeutic antibodies targeting prion proteins and β-amyloid peptides

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Prion diseases and Alzheimer’s disease (AD) are characterized by protein misfolding, and can lead to dementia. However, prion diseases are infectious and transmissible, while AD is not. The similarities and differences between these diseases have led researchers to perform comparative studies. In the last 2 decades, progress has been made in immunotherapy using anti-prion protein and anti-β-amyloid antibodies. In this study, we review new ideas and strategies for therapeutic antibodies targeting prion diseases and AD through conformation dependence.

  4. Hacking into the granuloma: could antibody antibiotic conjugates be developed for TB?

    Science.gov (United States)

    Ekins, Sean

    2014-12-01

    Alternatives to small molecule or vaccine approaches to treating tuberculosis are rarely discussed. Attacking Mycobacterium tuberculosis in the granuloma represents a challenge. It is proposed that the conjugation of small molecules onto a monoclonal antibody that recognizes macrophage or lymphocytes cell surface receptors, might be a way to target the bacteria in the granuloma. This antibody drug conjugate approach is currently being used in 2 FDA approved targeted cancer therapies. The pros and cons of this proposal for further research are discussed. PMID:25287628

  5. Detection of antibody responses by using haemagglutination inhibiton test and the protection titer of avian influenza virus H5N1 subtype

    Directory of Open Access Journals (Sweden)

    Risa Indriani

    2004-10-01

    Full Text Available Study on the detection of antibody responses using haemagglutination inhibition (HI test and the protection titer to Avian influenza (AI virus H5N1 subtype local isolate has been conducted at the Research Institute for Veterinary Science (RIVS. A total number of 50 village chicken (10 chicken served as un-injected controls and 30 quail were injected intramuscularly with inactivated virus of AI H5N1 subtype local isolate. Serum samples were collected 3 weeks after injection and were tested using haemagglutination inhibition tests. The correlation between antibody titer and its protection to AI virus H5N1 local isolate were measured by challenging the birds with AI virus H5N1 local isolate The HI test was then used to determine field serum samples. A total number of 48 village chicken from three (3 Districts (Bekasi, Tangerang and Bogor and 96 quails from two (2 farms in District of Sukabumi which were all vaccinated with commercial AI adjuvant vaccine were sampled. The study revealed that village chicken and quails showed antibody responses after 3 weeks vaccination and that titer of ≥ 3 log 2 was able to protect chicken and quails when they were challenged with local isolate virus. Based on this result, village chicken field samples from Districts of Tangerang, Bekasi and Bogor showed antibody titer which will protect 50, 100 and 85% of the flocks respectively. While quail field samples from Farm I and Farm II in District of Sukabumi showed antibody titer which will protect 60-100% and 0-80% of the flocks respectively. It is concluded that the study has successfully measured antibody titer to AI virus H5N1 subtype which protect village chicken and quails from local isolate virus challenge so that the results will be used to analyze field serum samples after vaccination program to eradicate AI from Indonesia.

  6. The meningococcal antibody test: how useful in the diagnosis of meningococcal disease?

    DEFF Research Database (Denmark)

    Weis, N; Berthelsen, L; Wachmann, H; Lind, I

    2005-01-01

    (29%) had been notified as cases of meningococcal disease. Among 228 patients notified as serologically confirmed the MAT results were consistent with the clinical diagnosis in 86%. MAT is a reliable tool for establishing a diagnosis in patients with suspected meningococcal disease. Key factors...... facilitating appropriate interpretation of negative as well as positive test results were: time(s) of sampling(s) after onset of disease, age of the patient and clinical features....

  7. Pulse Detonation Engine Test Bed Developed

    Science.gov (United States)

    Breisacher, Kevin J.

    2002-01-01

    A detonation is a supersonic combustion wave. A Pulse Detonation Engine (PDE) repetitively creates a series of detonation waves to take advantage of rapid burning and high peak pressures to efficiently produce thrust. NASA Glenn Research Center's Combustion Branch has developed a PDE test bed that can reproduce the operating conditions that might be encountered in an actual engine. It allows the rapid and cost-efficient evaluation of the technical issues and technologies associated with these engines. The test bed is modular in design. It consists of various length sections of both 2- and 2.6- in. internal-diameter combustor tubes. These tubes can be bolted together to create a variety of combustor configurations. A series of bosses allow instrumentation to be inserted on the tubes. Dynamic pressure sensors and heat flux gauges have been used to characterize the performance of the test bed. The PDE test bed is designed to utilize an existing calorimeter (for heat load measurement) and windowed (for optical access) combustor sections. It uses hydrogen as the fuel, and oxygen and nitrogen are mixed to simulate air. An electronic controller is used to open the hydrogen and air valves (or a continuous flow of air is used) and to fire the spark at the appropriate times. Scheduled tests on the test bed include an evaluation of the pumping ability of the train of detonation waves for use in an ejector and an evaluation of the pollutants formed in a PDE combustor. Glenn's Combustion Branch uses the National Combustor Code (NCC) to perform numerical analyses of PDE's as well as to evaluate alternative detonative combustion devices. Pulse Detonation Engine testbed.

  8. Design and development of CAMAC test module

    International Nuclear Information System (INIS)

    Various Computer automated measurement and control (CAMAC) modules are used in control and monitoring of Pelletron Accelerator. 24 channels CAMAC Input Gate is used for getting the ON/OFF status of various devices in the Pelletron Accelerator. If a channel has 24 V then the status is 'ON' and if the channel receives 0 V then the status is 'OFF'. Hence we can get the status of 24 different channels though one CAMAC Input Gate module. The status is transported to the PC via CAMAC controller. The manual testing of CAMAC Input Gate involves connection of 24 V to each channel and checking the status of each channel with Graphical user interface (GUI) software. This process of checking input gate is automated by developing a CAMAC Test module which is connected to CAMAC Input Gate with a 50 pin ribbon cable. The Test module automatically generates 24 V /0 V on each channel to be tested depending on the software GUI buttons labeled as 'ON'/'OFF' in labview. The status of CAMAC Input Gate is displayed on GUI for all 24 channels. Hence the user can check the working of each channel on GUI written in labview. This automated process of checking the CAMAC Input Gate saves time to debug problems in module and identifying the bad channel which can be subsequently repaired. The CAMAC Test module uses Spartan 2 FPGA which is connected to 24 transistors which in turn operates 24 relays. 24 V supply is connected to the relay secondary contacts which open/close as per the transistor inputs. The 24 V contacts are connected to the module output connector which should be connected to CAMAC Input Gate which is to be tested. (author)

  9. Development of a serotype colloidal gold strip using monoclonal antibody for rapid detection type Asia1 foot-and-mouth disease

    Directory of Open Access Journals (Sweden)

    Gao Shan-dian

    2011-09-01

    Full Text Available Abstract Background In this study, we developed a rapid, one step colloid gold strip (CGS capable of specifically detecting type Asia1 foot-and-mouth disease virus (FMDV. We have produced two monoclonal antibodies (mAb to type Asia1 FMD (named 1B8 and 5E2. On the test strip, the purified 1B8 labelled with the colloidal gold was used as the detector, and the purified 5E2 and goat anti-mouse antibodies were wrapped onto nitrocellulose (NC membranes as the test and the control line, respectively. The rapid colloidal gold stereotype diagnostic strip was housed in a plastic case. Results In specificity and sensitivity assay, there was no cross-reaction of the antigen with the other type of FMD and SVDV. The detection sensitivity was found to be as high as 10-5 dilution of Asia1/JSL/05 (1 × 107.2TCID50/50 μL. There was excellent agreement between the results obtained by CGS and reverse indirect hemagglutination assay (RIHA, and the agreement can reach to 98.75%. Conclusion We developed colloidal gold strips that have good qualities and does not require specialized equipment or technicians. This method provided a feasible, convenient, rapid, and effective for detecting type Asia1 FMDV in the fields.

  10. Progression of Mycoplasma hyosynoviae infection in three pig herds. Development of tonsillar carrier state, arthritis and antibodies in serum and synovial fluid in pigs from birth to slaughter

    DEFF Research Database (Denmark)

    Hagedorn-Olsen, T.; Nielsen, N.C.; Friis, N.F.; Nielsen, Jens

    1999-01-01

    In this investigation, natural infection with Mycoplasma hyosynoviae was followed in groups of individual pigs in three different herds with regard to occurrence of tonsillar carrier state, clinical arthritis and development of antibodies in serum and in synovial fluid. Antibodies were detected b...

  11. Antibody repertoire development in fetal and neonatal piglets. XXI. VH usage remains constant during development in fetal piglets and postnatally in piglets exposed to environmental antigen

    Science.gov (United States)

    Usage of variable region gene segments during development of the antibody repertoire in mammals is unresolved in part because of the complexity of the locus in mice and humans and the difficulty of distinguishing intrinsic from extrinsic influences in these species. We have addressed this using a hi...

  12. Clinical Validation of a Point-of-Care Multiplexed In Vitro Immunoassay Using Monoclonal Antibodies (the MSD Influenza Test) in Four Hospitals in Vietnam

    Science.gov (United States)

    van Kinh, Nguyen; Tuan, Ha Manh; Tuan, Tran Anh; Minh, Ngo Ngoc Quang; Bryant, Juliet E.; Hang, Vu thi Ty; Uyen, Le thi Tham; Thinh, Le Quoc; Anh, Tran thi Ngoc; Lan, Nguyen Phu Huong; Trung, Nguyen Vu; Taylor, Walter; Merson, Laura; Wertheim, Heiman F. L.; Farrar, Jeremy; Wolbers, Marcel; Chau, Nguyen van Vinh; de Jong, Menno D.

    2012-01-01

    Point-of-care (POC) diagnostic tests for influenza can considerably shorten the time to clinical decision making. An investigational POC test based on a multiplexed immunoassay was developed by Meso Scale Diagnostics, LLC (MSD), with the objective to make a more sensitive rapid test that can also subtype influenza A viruses (1977 H1, H3, and H5). Between February and November 2010, we conducted a prospective multicenter study at four hospitals in Vietnam and compared the performance of this test to that of the WHO/CDC real-time reverse transcriptase PCR (RT-PCR) on nasal and throat swab specimens from patients presenting with influenza-like illness. Five hundred sixty-three adults and children with a median age of 25 months were enrolled. Sensitivity and specificity of the test with combined results from nasal and throat swab samples were 74.0% (131/177) and 99.7% (351/352), respectively, compared to RT-PCR. The POC test was as sensitive for influenza virus B as for influenza virus A (74.4% [64/86] versus 73.6% [67/91]). The positivity rate was associated with lower cycle threshold values (a marker for higher viral loads), sample type (73.6% for nasal swab versus 52.4% for throat swab), and younger age. A total of 210 (18.7%) out of 1,126 MSD tests failed, and for 34 (6%) of patients, both test samples failed (these were excluded from the performance analysis). Subtyping could be assessed only for influenza virus A/H3N2, as 1977 H1N1 was not circulating at the time and no H5N1-infected patients were enrolled, and was successful only in 9/54 patients infected with H3 influenza virus who had a positive POC test result for influenza virus A. This novel POC test provided highly sensitive detection of influenza viruses A and B compared to the reported sensitivities of other rapid tests. However, 18.7% of tests failed for technical reasons and subtyping for H3 was poor. Drawbacks to the technology include the requirement for a dedicated reader instrument and the need for

  13. Development of monoclonal antibody against isoquinoline alkaloid coptisine and its application for the screening of medicinal plants

    OpenAIRE

    Kim, Jun-Sik; Tanaka, Hiroyuki; YUAN, CHUN-SU; Shoyama, Yukihiro

    2004-01-01

    In the development of immunoassay technique, the design of hapten containing a functional group suitable for protein conjugate is the key step for the preparation of antibodies against small molecules. Coptisine (MW 320), a bioactive constituent of Berberis and Coptis species, is small as an immunogen. In addition, coptisine has no reactive group in molecule for conjugating with a protein. To overcome this problem, 9-O-carboxymethyl-berberrubine was designed and conjugated with carrier protei...

  14. Development of a Specific Monoclonal Antibody-Based ELISA to Measure the Artemether Content of Antimalarial Drugs

    OpenAIRE

    Suqin Guo; Yongliang Cui; Lishan He; Liang Zhang; Zhen Cao; Wei Zhang; Rui Zhang; Guiyu Tan; Baomin Wang; Liwang Cui

    2013-01-01

    Artemether is one of the artemisinin derivatives that are active ingredients in antimalarial drugs. Counterfeit and substandard antimalarial drugs have become a serious problem, which demands reliable analytical tools and implementation of strict regulation of drug quality. Structural similarity among artemisinin analogs is a challenge to develop immunoassays that are specific to artemisinin derivatives. To produce specific antibodies to artemether, we used microbial fermentation of artemethe...

  15. Development and application of monoclonal antibodies for in situ detection of indigenous bacterial strains in aquatic ecosystems.

    OpenAIRE

    Faude, U C; Höfle, M.G.

    1997-01-01

    Strain-specific monoclonal antibodies (MAbs) were developed for three different bacterial isolates obtained from a freshwater environment (Lake Plusssee) in the spring of 1990. The three isolates, which were identified by molecular methods, were as follows: Cytophaga johnsonae PX62, Comamonas acidovorans PX54, and Aeromonas hydrophila PU7718. These strains represented three species that were detected in high abundance during a set of mesocosm experiments in Lake Plusssee by the direct analysi...

  16. A comparative study between Rose Bengal and indirect ELISA tests for detection of the antibrucella antibodies in serum of goats in Mosul city

    Directory of Open Access Journals (Sweden)

    S. H. Arslan

    2010-01-01

    Full Text Available This study was conducted to determine seroprevalence of brucellosis in non vaccinated goats in different areas in Mosulcity and to compare Rose Bengal and indirect ELISA tests for the detection of the antibodies. A total of 102 blood serumsamples were examined, representing 396 goats distributed in different areas. Results of Rose Bengal test showed that the totalseroprevalence of brucellosis was (6.8% and the highest rate in Al-Rahmania (11.1%, and lowest (7.1% in Al-Muthana area.No seroprevalence of Brucellosis was recorded in Googjaly and Basheka areas. Indirect ELISA test showed that the totalpercentage was (24.5% and the highest (55% were reported in Al-Rahmania area and the lowest percentage was (10% inBasheka area. The compatibility between the two tests was (0.30 on Kappa value indicating sensitivity of indirect ELISA testcompared with Rose Bengal test in detection antibrucella antibodies in serum goats.

  17. Monoclonal antibodies and cancer

    International Nuclear Information System (INIS)

    The usefulness of radiolabeled monoclonal antibodies for imaging and treatment of human (ovarian) cancer was investigated. A review of tumor imaging with monoclonal antibodies is presented. Special attention is given to factors that influence the localization of the antibodies in tumors, isotope choice and methods of radiolabeling of the monoclonal antibodies. Two monoclonal antibodies, OC125 and OV-TL3, with high specificity for human epithelial ovarian cancer are characterized. A simple radio-iodination technique was developed for clinical application of the monoclonal antibodies. The behavior of monoclonal antibodies in human tumor xenograft systems and in man are described. Imaging of tumors is complicated because of high background levels of radioactivity in other sites than the tumor, especially in the bloodpool. A technique was developed to improve imaging of human tumor xenographs in nude mice, using subtraction of a specific and a non-specific antibody, radiolabeled with 111In, 67Ga and 131I. To investigate the capability of the two monoclonal antibodies, to specifically localize in human ovarian carcinomas, distribution studies in mice bearing human ovarian carcinoma xenografts were performed. One of the antibodies, OC125, was used for distribution studies in ovarian cancer patients. OC125 was used because of availability and approval to use this antibody in patients. The same antibody was used to investigate the usefulness of radioimmunoimaging in ovarian cancer patients. The interaction of injected radiolabeled antibody OC125 with circulating antigen and an assay to measure the antibody response in ovarian cancer patients after injection of the antibody is described. 265 refs.; 30 figs.; 19 tabs

  18. New oligopeptide immunoglobulin G test for human parvovirus B19 antibodies.

    OpenAIRE

    Schwarz, T F; Modrow, Susanne; Hottenträger, B; Höflacher, B.; Jäger, G; Scharti, W.; Sumazakl, R.; Wolf, Hans J.; Middeldorp, J.; Roggendorf, M

    1991-01-01

    A new, highly sensitive and specific enzyme immunoassay using oligopeptides as antigen (enzyme-linked immunosorbent assay [ELISA] B19-OP) for detecting parvovirus B19-specific immunoglobulin G (IgG) was established. As antigens, B19-specific oligopeptides of 24 and 30 kDa derived from a 196-kDa fusion protein of beta-galactosidase and viral capsid protein (VPI) of B19 after CNBr cleavage and separation by high-pressure liquid chromatography were used. Of 139 serum specimens tested in parallel...

  19. Development of polyclonal antibodies against neurosteroids and their use in the immunoaffinity chromatography

    Czech Academy of Sciences Publication Activity Database

    Oklešťková, Jana; Pařízková, Barbora; Novák, Ondřej; Kapras, Vojtěch; Chodounská, Hana; Strnad, Miroslav

    2014-01-01

    Roč. 108, S2 (2014), s139. ISSN 0009-2770. [Conference on Isoprenoids /22./. 07.09.2014-10.09.2014, Praha] R&D Projects: GA MŠk(CZ) LO1204; GA MŠk LK21306 Institutional support: RVO:61388963 ; RVO:61389030 Keywords : neurosteroids * polyclonal antibodies Subject RIV: EF - Botanics

  20. Development of Re-188 radiolabelling procedures of peptides and monoclonal antibodies

    International Nuclear Information System (INIS)

    The goal of this work was to study and select the parameters to label biomolecules directly with 188Re. The concrete objectives pursued were: (i) Labelling h-IgG with 188Re; (ii) Radiolabelling Lanreotide and anti-CEA monoclonal antibody with 188Re; investigation of chromatography based quality control techniques; (iii) Biodistribution studies in tumour bearing rats

  1. Development of neutralizing monoclonal antibodies for oncogenic human papillomavirus types 31, 33, 45, 52, and 58.

    Science.gov (United States)

    Brown, Martha J; Seitz, Hanna; Towne, Victoria; Müller, Martin; Finnefrock, Adam C

    2014-04-01

    Human papillomavirus (HPV) is the etiological agent for all cervical cancers, a significant number of other anogenital cancers, and a growing number of head and neck cancers. Two licensed vaccines offer protection against the most prevalent oncogenic types, 16 and 18, responsible for approximately 70% of cervical cancer cases worldwide and one of these also offers protection against types 6 and 11, responsible for 90% of genital warts. The vaccines are comprised of recombinantly expressed major capsid proteins that self-assemble into virus-like particles (VLPs) and prevent infection by eliciting neutralizing antibodies. Adding the other frequently identified oncogenic types 31, 33, 45, 52, and 58 to a vaccine would increase the coverage against HPV-induced cancers to approximately 90%. We describe the generation and characterization of panels of monoclonal antibodies to these five additional oncogenic HPV types, and the selection of antibody pairs that were high affinity and type specific and recognized conformation-dependent neutralizing epitopes. Such characteristics make these antibodies useful tools for monitoring the production and potency of a prototype vaccine as well as monitoring vaccine-induced immune responses in the clinic. PMID:24574536

  2. Antineutrophil Cytoplasmic Antibodies Testing in a Large Cohort of Unselected Greek Patients

    Directory of Open Access Journals (Sweden)

    Konstantinos Tsiveriotis

    2011-01-01

    Full Text Available Objective. To retrospectively evaluate ANCA testing in a cohort of unselected Greek in- and outpatients. Methods. In 10803 consecutive serum samples, ANCA were tested by indirect immunofluorescence (IIF and ELISA. ELISA in inpatients was performed only on IIF positive sera. Results. Low prevalence (6.0% of IIF positive samples was observed. Among these samples, 63.5% presented perinuclear (p-ANCA, 9.3% cytoplasmic (c-ANCA and 27.2% atypical (x-ANCA pattern. 16.1% of p-ANCA were antimyeloperoxidase (anti-MPO positive, whereas 68.3% of c-ANCA were antiproteinase-3 (anti-PR3 positive. Only 17 IIF negative outpatients' samples were ELISA positive. ANCA-associated vasculitides (AAV, connective tissue disorders and gastrointestinal disorders represented 20.5%, 23.9%, and 21.2% of positive results, respectively. AAV patients exhibited higher rates of MPO/PR3 specificity compared to non-AAV (93.8% versus 8%. Conclusions. This first paper on Greek patients supports that screening for ANCA by IIF and confirming positive results by ELISA minimize laboratory charges without sacrificing diagnostic accuracy.

  3. Synthetic Antibodies for Reversible Cell Recognition

    Science.gov (United States)

    Zhou, Jing Zhou

    2011-12-01

    Antibody-mediated cell recognition plays a critical role in various biological and biomedical applications. However, strong antibody-cell interactions can lead to the difficulty of separating antibodies from the bound cells in a simple and non-destructive manner, which is often necessary to numerous applications such as cell sorting or separation. Thus, this thesis research is aimed to create an antibody-like nanomaterial with the function of reversible cell recognition It was hypothesized that nucleic acid aptamer and dendrimer could be used as fundamental structural components to develop an antibody-like nanomaterial. The aptamer functions as the binding site of an antibody; the dendrimer is used as a robust, defined nano-scaffold to support the aptamer and to carry small molecules (e.g., fluorophores). To test this hypothesis, a novel method was first developed to discover the essential nucleotides of full-length aptamers to mimic the binding sites of antibodies. The essential nucleotides were further conjugated with a dendrimer to synthesize a monovalent aptamer-dendrimer nanomaterial. The results clearly showed that the essential nucleotides could maintain high affinity and specificity after tethered on dendrimer surface. To further test the hypothesis that antibody-like nanomaterials can be rationally designed to acquire the capability of reversible cell recognition, an aptamer that was selected at 0 °C was used as a model to synthesize a "Y-shaped" nanomaterial by conjugating two aptamers to the same dendrimer. The results showed that the nanomaterial-cell interaction could be affected by the distance between two binding aptamers. In addition, the "Y-shaped" antibody-like nanomaterial could bind target cells more strongly than its monovalent control. Importantly, the strong cell-nanomaterial interaction could be rapidly reversed when the temperature was shifted from 0 °C to 37 °C. In summary, we developed a synthetic antibody that can not only mimic the

  4. Penetration Testing Curriculum Development in Practice

    Directory of Open Access Journals (Sweden)

    Chengcheng Li

    2015-04-01

    report for the management team to aid in strengthening the system, never to cause any real damages. This paper introduces the development of a penetration testing curriculum as a core class in an undergraduate cybersecurity track in Information Technology. The teaching modules are developed based on the professional penetration testing life cycle. The concepts taught in the class are enforced by hands-on lab exercises. This paper also shares the resources that are available to institutions looking for teaching materials and grant opportunities to support efforts when creating a similar curriculum in cybersecurity.

  5. Progress in the development of immunoanalytical methods incorporating recombinant antibodies to small molecular weight biotoxins.

    Science.gov (United States)

    Kavanagh, Owen; Elliott, Christopher T; Campbell, Katrina

    2015-04-01

    Rapid immunoanalytical screening of food and environmental samples for small molecular weight (hapten) biotoxin contaminations requires the production of antibody reagents that possess the requisite sensitivity and specificity. To date animal-derived polyclonal (pAb) and monoclonal (mAb) antibodies have provided the binding element of the majority of these assays but recombinant antibodies (rAb) isolated from in vitro combinatorial phage display libraries are an exciting alternative due to (1) circumventing the need for experimental animals, (2) speed of production in commonly used in vitro expression systems and (3) subsequent molecular enhancement of binder performance. Short chain variable fragments (scFv) have been the most commonly employed rAb reagents for hapten biotoxin detection over the last two decades but antibody binding fragments (Fab) and single domain antibodies (sdAb) are increasing in popularity due to increased expression efficiency of functional binders and superior resistance to solvents. rAb-based immunochromatographic assays and surface plasmon resonance (SPR) biosensors have been reported to detect sub-regulatory levels of fungal (mycotoxins), marine (phycotoxins) and aquatic biotoxins in a wide range of food and environmental matrices, however this technology has yet to surpass the performances of the equivalent mAb- and pAb-based formats. As such the full potential of rAb technology in hapten biotoxin detection has yet to be achieved, but in time the inherent advantages of engineered rAb are set to provide the next generation of ultra-high performing binder reagents for the rapid and specific detection of hapten biotoxins. PMID:25716465

  6. A solid-phase radioimmunoassay for detection of tetanus antibodies

    International Nuclear Information System (INIS)

    A solid-phase radioimmunoassay has been developed as a screening technique for tetanus antibodies in blood plasma. It is based on the principle of a commercial test for Hepatitis B antibody. Compared to previous screening techniques, the radioimmunoassay showed better stability with no apparent loss of sensitivity over a 2 month period. This technique has proved useful in determining tetanus immunity and in monitoring free antibody level in treated cases of clinical tetanus. (U.K.)

  7. Steady reconstruction process - development, testing and comparison in ultrasonic testing

    International Nuclear Information System (INIS)

    The fault parameters can be extracted from a few data of high quality in steady test procedures. The boundary conditions for the successful use of such a process were researched and found, so that by using theoretical models for the elasto-dynamic interaction of fault and ultrasonics, a concentration of wavefronts instead of resonances and a wide band careful collection of data makes a physical interpretation in the form of specific geometry torques possible. Models of the interaction of ultrasonics and faults for two fault geometries (cracks and pores) were developed which permit the calculation of A scans of any bandwidth and with any angle of scatter for the direct and mode converted parts of the elastic ultrasonic scatter wave. The curved pressure and shear waves including the mode converted bending fields over an angular range of 360deg were experimentally recorded. Their agreement including the additional wavefronts caused by the close field of the crack bending field is close. Classification of torques is done on two examples (crack, cylinder) for evaluation purposes. It was found that a classification was possible according to the sign of the a1 polynomial coefficient. (orig./HP)

  8. Developing the MAPLE materials test reactor concept

    International Nuclear Information System (INIS)

    MAPLE-MTR is a new multipurpose research facility being planned by AECL Research as a possible replacement for the 35-year-old NRU reactor. In developing the MAPLE-MTR concept, AECL is starting from the recent design and licensing experience with the MAPLE-X10 reactor. By starting from technology developed to support the MAPLE-X10 design and adapting it to produce a concept that satisfies the requirements of fuel channel materials testing and fuel irradiation programs, AECL expects to minimize the need for major advances in nuclear technology (e.g., fuel, heat transfer). Formulation of the MAPLE-MTR concept is at an early stage. This report describes the irradiation requirements of the research areas, how these needs are translated into design criteria for the project and elements of the preliminary design concept

  9. Monoclonal antibodies to nerve growth factor affect the postnatal development of the visual system.

    OpenAIRE

    N.Berardi; Cellerino, A.; L. DOMENICI; Fagiolini, M.; Pizzorusso, T.; Cattaneo, A.; L Maffei

    1994-01-01

    Exogenous supply of nerve growth factor (NGF) prevents the effects of monocular deprivation. This suggests that visual afferents may be competing for an endogenous neurotrophic factor, related to NGF, whose production by postsynaptic cells depends on the activity of afferent fibers. To test the hypothesis that endogenous NGF may play a role in the functional and anatomical development of the rat geniculo cortical system, the physiological action of NGF in the rat visual system was antagonized...

  10. The value of non-human primates in the development of therapeutic monoclonal antibodies

    OpenAIRE

    Van Meer, P.J.K.; Kooijman, M.; Van Der Laan, J.W.; Moors, E.H.M.; Schellekens, H

    2011-01-01

    The pharmaceutical industry is increasingly focusing on the development of biological therapeutics. These molecules generally cause no off-target toxicity and are highly species specific. Therefore, non-human primates (NHPs) are often the only relevant species in which to conduct regulatory safety testing to support clinical trials. However, species specificity and immunogenicity may negatively impact the predictive value of these ethically contentious animals and thus limits their value as a...

  11. Detection of polysaccharide antigens of Candida albicans interfering with specific antibodies in human sera

    International Nuclear Information System (INIS)

    A double antibody sandwich radioimmunoassay was developed for the detection of circulating polysaccharide antigens of Candida albicans. The sensitivity of the assay for polysaccharides was 1 ng/mL. The in-vitro interference of specific polysaccharide antibodies, even from sera with low antibody levels, could be demonstrated. The sensitivity of the antigen detection decreased proportionally to the amount of polysaccharide antibodies in the sera. The sensitivity of the assay was almost completely restored by heating the sera. This procedure destroyed antibodies and the released polysaccharide antigens were detectable in the test system by using radiolabelled anti-polysaccharide antibodies. (author)

  12. Review for the generalist: The antinuclear antibody test in children - When to use it and what to do with a positive titer

    OpenAIRE

    Sailer-Hoeck Michaela; Mackinnon Murray J; Malleson Peter N; Spencer Charles H

    2010-01-01

    Abstract The antinuclear antibody test (ANA) is a much overused test in pediatrics. The ANA does have a role in serologic testing but it should be a very limited one. It is often ordered as a screening test for rheumatic illnesses in a primary care setting. However, since it has low specificity and sensitivity for most rheumatic and musculoskeletal illnesses in children, it should not be ordered as a screening test for non-specific complaints such as musculoskeletal pain. It should only be us...

  13. Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the herbicide chlorimuron-ethyl.

    Science.gov (United States)

    Zhao, Jing; Yi, Guo-Xiang; He, Su-Ping; Wang, Bao-Min; Yu, Cai-Xia; Li, Gang; Zhai, Zhi-Xi; Li, Zhao-Hu; Li, Qing X

    2006-07-12

    Hybridomas secreting a monoclonal antibody (mAb) against the herbicide chlorimuron-ethyl (CE) were produced by fusing the mouse myeloma cell line (SP2/0) with splenocytes from a mouse immunized against the conjugate of the sulfonamide moiety of CE and bovine serum albumin (BSA). The mAb, designated 1F5C5A10, had very weak affinity with metsulfuron, ethametsulfuron, pyrazosulfuron, bensulfuron, and chlorsulfuron. Two mAb-based indirect competitive enzyme-linked immunosorbent assays (icELISA) were developed. A conventional icELISA (icELISA-I) showed a concentration of half-maximum inhibition (IC(50)) of 11.6 ng/mL with a dynamic range of 1.6-84 ng/mL. A simplified icELISA (icELISA-II) had an IC(50) of 28.7 ng/mL and a dynamic range of 2.2-372 ng/mL. The two assays were tested on spiked water and soil samples. CE (1-500 ng/mL) fortified in water samples could be analyzed directly without any sample preparation by both immunoassays with an average recovery between 74 and 114%. icELISA-II, but not icELISA-I, was able to accurately analyze the herbicide residues in the crude soil extracts with recoveries between 99 and 129% without obvious matrix effects due to its lesser amount of sample used. In contrast to icELISA-I, icELISA-II is more convenient, whereas it consumes more reagents of coating antigen and goat anti-mouse IgG-peroxidase. PMID:16819901

  14. Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of "preneoplastic antigen"-like molecules.

    Science.gov (United States)

    Duan, Hongying; Yoshimura, Kazunori; Kobayashi, Nobuharu; Sugiyama, Kazuo; Sawada, Jun-Ichi; Saito, Yoshiro; Morisseau, Christophe; Hammock, Bruce D; Akatsuka, Toshitaka

    2012-04-01

    Microsomal epoxide hydrolase (mEH) is a drug metabolizing enzyme which resides on the endoplasmic reticulum (ER) membrane and catalyzes the hydration of reactive epoxide intermediates that are formed by cytochrome P450s. mEH is also thought to have a role in bile acid transport on the plasma membrane of hepatocytes. It is speculated that efficient execution of such multiple functions is secured by its orientation and association with cytochrome P450 enzymes on the ER membrane and formation of a multiple transport system on the plasma membrane. In certain disease status, mEH loses its association with the membrane and can be detected as distinct antigens in the cytosol of preneoplastic foci of liver (preneoplastic antigen), in the serum in association with hepatitis C virus infection (AN antigen), or in some brain tumors. To analyze the antigenic structures of mEH in physiological and pathological conditions, we developed monoclonal antibodies against different portions of mEH. Five different kinds of antibodies were obtained: three, anti-N-terminal portions; one anti-C-terminal; and one, anti-conformational epitope. By combining these antibodies, we developed antigen detection methods which are specific to either the membrane-bound form or the linearized form of mEH. These methods detected mEH in the culture medium released from a hepatocellular carcinoma cell line and a glioblastoma cell line, which was found to be a multimolecular complex with a unique antigenic structure different from that of the membrane-bound form of mEH. These antibodies and antigen detection methods may be useful to study pathological changes of mEH in various human diseases. PMID:22310175

  15. Detection of filarial specific IgG4 antibodies in individuals residing in endemic areas using panLFRAPID test card

    OpenAIRE

    K Athisaya Mary; Hoti, S. L.; Krishnamoorthy, K.; Das, P K; Rahmah, N.

    2011-01-01

    In order to achieve the goal of global programme for elimination of lymphatic filariasis (GPELF), chemotherapy programmes are underway to interrupt transmission of the disease. At this point, detection of exposure will be more appropriate to monitor the programme and to certify areas cleared of active transmission as disease-free. A recently available cassette form of rapid test, panLFRAPID is a filarial IgG4 antibody detection test that may be useful for the programme. Therefore, we carried ...

  16. Seroprevalence of Antibodies to Bartonella henselae in Patients with Cat Scratch Disease and in Healthy Controls: Evaluation and Comparison of Two Commercial Serological Tests

    OpenAIRE

    Sander, Anna; Posselt, Miriam; Oberle, Karin; Bredt, Wolfgang

    1998-01-01

    Serologic testing for the presence of antibodies to Bartonella henselae is a widely accepted diagnostic procedure for laboratory confirmation of the diagnosis of cat scratch disease (CSD). In this study a commercially available indirect immunofluorescence assay (IFA) based on B. henselae-infected human larynx carcinoma cells (test A) was evaluated. Sera from 42 patients with CSD (20 confirmed by PCR) and 270 sera from healthy controls (consisting of 63 cat owners, 65 individuals whose last cl...

  17. Frequencies and Specificities of “Enzyme-Only” Detected Erythrocyte Alloantibodies in Patients Hospitalized in Austria: Is an Enzyme Test Required for Routine Red Blood Cell Antibody Screening?

    Directory of Open Access Journals (Sweden)

    Dietmar Enko

    2014-01-01

    Full Text Available The aim of this study was to determine the frequencies and specificities of “enzyme-only” detected red blood cell (RBC alloantibodies in the routine antibody screening and antibody identification in patients hospitalized in Austria. Routine blood samples of 2420 patients were investigated. The antibody screening was performed with a 3-cell panel in the low-ionic strength saline- (LISS- indirect antiglobulin test (IAT and with an enzyme-pretreated (papain 3-cell panel fully automated on the ORTHO AutoVue Innova System. The antibody identification was carried out manually with an 11-cell panel in the LISS-IAT and with an enzyme-pretreated (papain 11-cell panel. In total 4.05% (n=98 of all patients (n=2420 had a positive RBC antibody screening result. Of them 25.51% (25/98 showed “enzyme-only” detected specific or nonspecific RBC alloantibodies. Rhesus and Lewis system antibodies were found the only specificities of “enzyme-only” RBC alloantibodies: all in all 4.8% (4/98 were detected with anti-E, 3.06% (3/98 with anti-Lea, 3.06% (3/98 with anti-D after anti-D prophylaxis and 1.02% (1/98 with anti-e. In total, 14.29% (14/98 showed a nonspecific RBC alloantibody result with the enzyme test. The results of the present study demonstrate that a high number of unwanted positive reactions with the enzyme technique overshadows the detection of “enzyme-only” RBC alloantibodies. (Trial Registration: K-37-13.

  18. Development of rast assay for determination of anti-popolus canadensis IgE antibodies

    International Nuclear Information System (INIS)

    One of the frequent causes of pollen allergy in our region (Serbia, Yugoslavia) is the pollen of poplar (Populus canadensis). The aim of this study was to form RAST for the determination of specific anti-Populus canadensis IgE antibodies. Affinity purified and radiolabelled (I-125) MoAb E1 was used for forming assay for the determination of specific IgE. By titration of extract of poplar Populus canadensis we determined that the quantity of 0.65 mL extract is needed for coupling of Ig BrCN activated paper discs. Coupling was performed in Na2CO3/NaHCO3 buffer pH 10, on 4 deg C for 48h. Using this newly formed RAST, specific for Populus canadensis, we have determinated anti-Populus canadensis IgE antibodies as well as cross reactivity between pollens of Populus canadensis and Populus deltoides. (author). 12 refs., 3 figs., 1 tab

  19. Clinical development methodology for infusion-related reactions with monoclonal antibodies

    OpenAIRE

    Doessegger, Lucette; Banholzer, Maria Longauer

    2015-01-01

    Infusion-related reactions (IRRs) are common with monoclonal antibodies (mAbs) and timely related to drug administration and have been reported as anaphylaxis, anaphylactoid reactions and cytokine release syndrome, among other terms used. We address risk management measures for individual patients and for the study and propose a consistent reporting approach in an attempt to allow cross-molecule comparisons. Once the symptoms of IRR have resolved, the mAb may be restarted. Rechallenge should ...

  20. Rational development of high-affinity T-cell receptor-like antibodies.

    Science.gov (United States)

    Stewart-Jones, Guillaume; Wadle, Andreas; Hombach, Anja; Shenderov, Eugene; Held, Gerhard; Fischer, Eliane; Kleber, Sascha; Nuber, Natko; Stenner-Liewen, Frank; Bauer, Stefan; McMichael, Andrew; Knuth, Alexander; Abken, Hinrich; Hombach, Andreas A; Cerundolo, Vincenzo; Jones, E Yvonne; Renner, Christoph

    2009-04-01

    T-cell interaction with a target cell is a key event in the adaptive immune response and primarily driven by T-cell receptor (TCR) recognition of peptide-MHC (pMHC) complexes. TCR avidity for a given pMHC is determined by number of MHC molecules, availability of coreceptors, and TCR affinity for MHC or peptide, respectively, with peptide recognition being the most important factor to confer target specificity. Here we present high-resolution crystal structures of 2 Fab antibodies in complex with the immunodominant NY-ESO-1(157-165) peptide analogue (SLLMWITQV) presented by HLA-A*0201 and compare them with a TCR recognizing the same pMHC. Binding to the central methionine-tryptophan peptide motif and orientation of binding were almost identical for Fabs and TCR. As the MW "peg" dominates the contacts between Fab and peptide, we estimated the contributions of individual amino acids between the Fab and peptide to provide the rational basis for a peptide-focused second-generation, high-affinity antibody library. The final Fab candidate achieved better peptide binding by 2 light-chain mutations, giving a 20-fold affinity improvement to 2-4 nM, exceeding the affinity of the TCR by 1,000-fold. The high-affinity Fab when grafted as recombinant TCR on T cells conferred specific killing of HLA-A*0201/NY-ESO-1(157-165) target cells. In summary, we prove that affinity maturation of antibodies mimicking a TCR is possible and provide a strategy for engineering high-affinity antibodies that can be used in targeting specific pMHC complexes for diagnostic and therapeutic purposes. PMID:19307587

  1. Rapid HIV testing for developing countries: the challenge of false-negative tests

    Science.gov (United States)

    Yogev, Ram

    2012-06-01

    It is a common practice in resource-constrained countries to accept two positive rapid HIV antibody test results as diagnostic for HIV infection. Because these tests are inexpensive and results are obtained quickly, they are recommended by the WHO to "scale-up" HIV testing to increase the number of people tested. The negative predictive value of rapid HIV tests is so high that negative results are considered conclusive despite the fact that false-negative results can occur in several situations. While the specificity and sensitivity of rapid HIV tests in resource-rich countries is acceptable, there are only limited data about their performance in resource-constrained countries. The challenges of rapid HIV testing in these situations will be discussed.

  2. Development of a high-performance immunolatex based on "soft landing" antibody immobilization mechanism.

    Science.gov (United States)

    Yuan, Xiaofei; Fabregat, Dolça; Yoshimoto, Keitaro; Nagasaki, Yukio

    2012-11-01

    Rabbit anti-human ferritin (anti-hFT) polyclonal immunoglobulin G (IgG) and poly(ethylene glycol) (PEG) were sequentially co-immobilized onto polystyrene submicroparticles (sMPs) to construct sMP/anti-hFT/PEG (SAP) immunolatex. Chemical immobilization of anti-hFT was performed at different pH levels to evaluate variations in antigen recognition. Basic pH disfavored conjugation of anti-hFT to sMPs, but remarkably increased its antigen recognition in comparison to that at neutral pH. We investigated this intriguing phenomenon further by assessing the kinetics of antibody binding, including the time-dependency of immobilization, antigen recognition, and orientation of bound anti-hFT. Therefore, we attributed high antigen recognition to significant electrostatic repulsion between sMPs and anti-hFT at basic pH, which predominately prevented anti-hFT access to sMPs and concurrently promoted anti-hFT orientations suitable for antigen recognition. Subsequent PEG modification maintained such anti-hFT orientation, without which antigen-accessible orientations would have decreased with time. Thus, properly oriented antibody and immediate PEGylation after antibody immobilization contributed to the formation of a high-performance SAP immunolatex. PMID:22005261

  3. Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of “preneoplastic antigen”-like molecules

    International Nuclear Information System (INIS)

    Microsomal epoxide hydrolase (mEH) is a drug metabolizing enzyme which resides on the endoplasmic reticulum (ER) membrane and catalyzes the hydration of reactive epoxide intermediates that are formed by cytochrome P450s. mEH is also thought to have a role in bile acid transport on the plasma membrane of hepatocytes. It is speculated that efficient execution of such multiple functions is secured by its orientation and association with cytochrome P450 enzymes on the ER membrane and formation of a multiple transport system on the plasma membrane. In certain disease status, mEH loses its association with the membrane and can be detected as distinct antigens in the cytosol of preneoplastic foci of liver (preneoplastic antigen), in the serum in association with hepatitis C virus infection (AN antigen), or in some brain tumors. To analyze the antigenic structures of mEH in physiological and pathological conditions, we developed monoclonal antibodies against different portions of mEH. Five different kinds of antibodies were obtained: three, anti-N-terminal portions; one anti-C-terminal; and one, anti-conformational epitope. By combining these antibodies, we developed antigen detection methods which are specific to either the membrane-bound form or the linearized form of mEH. These methods detected mEH in the culture medium released from a hepatocellular carcinoma cell line and a glioblastoma cell line, which was found to be a multimolecular complex with a unique antigenic structure different from that of the membrane-bound form of mEH. These antibodies and antigen detection methods may be useful to study pathological changes of mEH in various human diseases. -- Highlights: ► Monoclonal antibodies against different portions of mEH were developed. ► They discriminate between the membrane-bound and the linearized forms of mEH. ► We analyze the antigenic structure of the altered form of mEH in tumor cells. ► Preneoplastic antigen is a multimolecular complex of mEH with

  4. Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of “preneoplastic antigen”-like molecules

    Energy Technology Data Exchange (ETDEWEB)

    Duan, Hongying [Department of Microbiology, Faculty of Medicine, Saitama Medical University, Moroyama-cho, Iruma-gun, Saitama 350-0495 (Japan); Yoshimura, Kazunori [Department of Physiology, Faculty of Medicine, Saitama Medical University, Moroyama-cho, Iruma-gun, Saitama 350-0495 (Japan); Kobayashi, Nobuharu; Sugiyama, Kazuo [Department of Microbiology, Faculty of Medicine, Saitama Medical University, Moroyama-cho, Iruma-gun, Saitama 350-0495 (Japan); Sawada, Jun-ichi; Saito, Yoshiro [Division of Biochemistry and Immunochemistry, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501 (Japan); Morisseau, Christophe; Hammock, Bruce D. [Department of Entomology and Cancer Center, University of California, Davis, One Shields Avenue, Davis, CA 95616-8584 (United States); Akatsuka, Toshitaka, E-mail: akatsuka@saitama-med.ac.jp [Department of Microbiology, Faculty of Medicine, Saitama Medical University, Moroyama-cho, Iruma-gun, Saitama 350-0495 (Japan)

    2012-04-01

    Microsomal epoxide hydrolase (mEH) is a drug metabolizing enzyme which resides on the endoplasmic reticulum (ER) membrane and catalyzes the hydration of reactive epoxide intermediates that are formed by cytochrome P450s. mEH is also thought to have a role in bile acid transport on the plasma membrane of hepatocytes. It is speculated that efficient execution of such multiple functions is secured by its orientation and association with cytochrome P450 enzymes on the ER membrane and formation of a multiple transport system on the plasma membrane. In certain disease status, mEH loses its association with the membrane and can be detected as distinct antigens in the cytosol of preneoplastic foci of liver (preneoplastic antigen), in the serum in association with hepatitis C virus infection (AN antigen), or in some brain tumors. To analyze the antigenic structures of mEH in physiological and pathological conditions, we developed monoclonal antibodies against different portions of mEH. Five different kinds of antibodies were obtained: three, anti-N-terminal portions; one anti-C-terminal; and one, anti-conformational epitope. By combining these antibodies, we developed antigen detection methods which are specific to either the membrane-bound form or the linearized form of mEH. These methods detected mEH in the culture medium released from a hepatocellular carcinoma cell line and a glioblastoma cell line, which was found to be a multimolecular complex with a unique antigenic structure different from that of the membrane-bound form of mEH. These antibodies and antigen detection methods may be useful to study pathological changes of mEH in various human diseases. -- Highlights: ► Monoclonal antibodies against different portions of mEH were developed. ► They discriminate between the membrane-bound and the linearized forms of mEH. ► We analyze the antigenic structure of the altered form of mEH in tumor cells. ► Preneoplastic antigen is a multimolecular complex of mEH with

  5. Development of monoclonal antibody-based galactomannoprotein antigen-capture ELISAs to detect Aspergillus fumigatus infection in the invasive aspergillosis rabbit models.

    Science.gov (United States)

    Wang, Z-Y; Cai, J-P; Qiu, L-W; Hao, W; Pan, Y-X; Tung, E T K; Lau, C C Y; Woo, P C Y; Lau, S K P; Yuen, K-Y; Che, X-Y

    2012-11-01

    Aspergillus fumigatus is one of the most prominent opportunistic fungal pathogens in immunocompromised hosts. Early recognition of this infection along with prompt antifungal therapy may increase the survival rate. We expressed two potential bio-markers of A. fumigatus infection-galactomannoprotein Afmp1p and Afmp4p in Pichia pastoris. We generated 33 monoclonal antibodies (MAbs), 20 against recombinant Afmp1p (rAfmp1p) and the other 13 against recombinant Afmp4p (rAfmp4p). Subsequently, we developed two antigen-capture enzyme-linked immunosorbent assays (ELISAs) which employed MAbs as both the capture and the detection antibodies for rAfmp1p and rAfmp4p. The two antigen-capture ELISAs specifically detected Afmp1p/Afmp4p in cultures of A. fumigatus and had no cross-reaction with other tested pathogenic fungi, including Penicillium marneffei and other pathogenic Aspergillus species. The Afmp1p-captured ELISA would be positive even when the culture supernatant of A. fumigatus had been diluted to 128-fold of its original concentration. The two antigen ELISAs could capture circulating or excreted antigens during the acute phase of invasive aspergillosis (IA) in the animal model, and had no cross-reactivity to other Aspergillus-challenged animal models. We developed two antigen-capture ELISAs for the laboratory diagnosis of A. fumigatus infection. These two antigen-capture ELISAs may be useful in the clinical diagnosis of aspergillosis. PMID:22669560

  6. "Unconventional" Neutralizing Activity of Antibodies Against HIV

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Neutralizing antibodies are recognized to be one of the essential elements of the adaptive immune response that must be induced by an effective vaccine against HIV. However, only a limited number of antibodies have been identified to neutralize a broad range of primary isolates of HIV-1 and attempts to induce such antibodies by immunization were unsuccessful. The difficulties to generate such antibodies are mainly due to intrinsic properties of HIV-1 envelope spikes, such as high sequence diversity, heavy glycosylation, and inducible and transient nature of certain epitopes. In vitro neutralizing antibodies are identified using "conventional" neutralization assay which uses phytohemagglutinin (PHA)-stimulated human PBMCs as target cells. Thus, in essence the assay evaluates HIV-1 replication in CD4+ T cells. Recently, several laboratories including us demonstrated that some monoclonal antibodies and HIV-1-specific polyclonal IgG purified from patient sera, although they do not have neutralizing activity when tested by the "conventional" neutralization assay, do exhibit potent and broad neutralizing activity in "unconventional" ways. The neutralizing activity of these antibodies and IgG fractions is acquired through post-translational modifications, through opsonization of virus particles into macrophages and immature dendritic cells (iDCs), or through expression of antibodies on the surface of HIV-1-susceptible cells. This review will focus on recent findings of this area and point out their potential applications in the development of preventive strategies against HIV.

  7. Development of a multiplexed bead-based immunoassay for the simultaneous detection of antibodies to 17 pneumococcal proteins.

    Science.gov (United States)

    Shoma, S; Verkaik, N J; de Vogel, C P; Hermans, P W M; van Selm, S; Mitchell, T J; van Roosmalen, M; Hossain, S; Rahman, M; Endtz, H Ph; van Wamel, W J B; van Belkum, A

    2011-04-01

    Presently, several pneumococcal proteins are being evaluated as potential vaccine candidates. Here, we gather novel insights in the immunogenicity of PLY, PsaA, PspA, PspC, NanA, Hyl, PpmA, SlrA, Eno, IgA1-protease, PdBD, BVH-3, SP1003, SP1633, SP1651, SP0189 and SP0376. We developed a multiplex bead-based immunoassay (xMAP(®) Technology, Luminex Corporation) to simultaneously quantify antibodies against these 17 pneumococcal proteins in serum. The median fluorescence intensity (MFI) values obtained for human pooled serum with the multiplex assay were between 82% and 111% (median 94%) of those obtained with the singleplex assays. For IgG, the coefficient of variation (CV) in serum ranged from 2% to 9%, for IgA, the CV ranged from 3% to 14% and for IgM, the CV ranged from 11% to 15%. Using this immunoassay, we showed that anti-pneumococcal antibody levels exhibited extensive inter-individual variability in young children suffering from invasive pneumococcal disease. All proteins, including the proteins with, as yet, unknown function, were immunogenic. In conclusion, the multiplex Streptococcus pneumoniae immunoassay based on proteins is reproducible. This assay can be used to monitor anti-S. pneumoniae antibody responses in a material- and time-saving manner. PMID:21086008

  8. RBC Antibody Screen

    Science.gov (United States)

    ... the baby is Rh-positive and the mother's antibody status is negative for anti-D, the mother is given additional RhIG. This test also may be used to help diagnose autoimmune-related hemolytic anemia ... when a person produces antibodies against his or her own RBC antigens. This ...

  9. Development of a monoclonal antibody-based broad-specificity ELISA for fluroquinolone antibiotics in foods and molecular modeling studies of cross-reactive compounds

    Science.gov (United States)

    Development of a competitive indirect enzyme-linked immunosorbent assay (ciELISA) with monoclonal antibodies (Mabs) having broad specificity for fluoroquinolone (FQ) antibiotics is described. Four FQs, ciprofloxacin (CIP), norfloxacin (NOR), enrofloxacin (ENR) and ofloxacin (OFL) were conjugated to...

  10. Avian reovirus antibody assay by indirect immunofluorescence using plastic microculture plates.

    OpenAIRE

    Ide, P R

    1982-01-01

    An indirect fluorescent antibody test was developed to detect serum antibody to avian reovirus strain WVU2937. This test employed small multiple well plastic plates (8 x 5.5 cm) which readily fitted into the standard mechanical stage mechanism of an incident light fluorescence microscope. The small wells of the plates required minimal (10 muL) volumes of reagents. In tests on 18 sera in which the indirect fluorescent antibody, agar gel precipitin and plaque reduction methods were compared ser...

  11. Serologic test systems development. Progress report, October 1, 1978-September 30, 1979

    Energy Technology Data Exchange (ETDEWEB)

    Seawright, G.L.; Sanders, W.M.; Hollstein, U.; Butler, J.E.; Mills, K.W.; Despommier, D.D.; Zimmerman, W.J.; Martinez, E.; Hindman, K.R.; Payne, R.J.

    1980-12-01

    Work has continued on the development and automation of enzyme immunoassays (EIA) for detecting diseases and toxic agents in food animals. Further evaluations were made of the Technicon Autoanalyzer II(AAII) for conducting totally automated EIAs. The problems investigated were machine carryover and assay variation. Modifications greatly reduced or eliminated carryover and produced acceptable levels of test variation. The EIA for swine trichinosis was significantly improved by the use of a new, partially purified antigen preparation. The result was improved detection of early seroconversions and reduced probability for false negatives and false positives. The amplified EIA was adapted as a diagnostic test for bovine brucellosis and studies were initiated for differentiating vaccinated and infected animals. Preliminary data indicate that the IgG/sub 1/ response may be diagnostic but further studies are necessary. Development of the EIA for detecting low molecular weight contaminants and residues in food products was also initiated. Compounds studied were the antibiotics chloramphenicol, tetracycline, and gentamicin; the mycotoxin, aflatoxin, and the shale oil toxin, 2-aminofluorene. Results indicate that chloramphenicol nonspecifically binds to antibody and interferes with antibody activity. Thus, the test is not yet satisfactory. Initial attempts to automate the gentamicin test were unsuccessful because of machine carryover but modifications of the AAII have produced encouraging preliminary data. Work is continuing on the development of EIAs for all of the compounds mentioned above. (ERB)

  12. Comparative Evaluation of Native Antigens for the Development of Brucellosis Antibody Detection System

    Directory of Open Access Journals (Sweden)

    Yasmin Bano

    2015-09-01

    Full Text Available Brucellosis is a highly infectious zoonotic disease and an economically important infection of humans and livestock with a worldwide distribution. The main mode of transmission of this disease to humans is through the consumption of infected milk, milk products, and uncooked or raw meat. The present study was designed to prepare few native antigens, that is, sonicated antigen (SA, cell envelope (CE antigen, and freeze and thaw (FT antigen from Brucella abortus S99 culture and to test them in a highly sensitive and specific indirect enzyme-linked immunosorbent assay (I-ELISA in both a microtiter plate and a dot-blot format for the development of field-based diagnosis. All 50 suspected bovine samples were tested by plate as well as in dot ELISA formats for all the three antigens prepared. The CE antigen was found to be more suitable as it had the maximum agreement with the Rose Bengal plate agglutination test results followed by the SA and the least agreement was found with that of the FT antigen. This detection system in microtiter plates and a dot-blot format will be useful for the rapid screening of samples for the disease surveillance and routine diagnosis.

  13. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    Science.gov (United States)

    Mohan, Anju; Saxena, Hari Mohan; Malhotra, Puneet

    2016-01-01

    Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT), microtiter plate agglutination test (MAT), indirect hemagglutination assay (IHA), and indirect enzyme-linked immunosorbent assay (iELISA) as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (pBrucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. PMID:27536032

  14. Production of a broad specificity antibody for the development and validation of an optical SPR screening method for free and intracellular microcystins and nodularin in cyanobacteria cultures.

    Science.gov (United States)

    Devlin, Shauna; Meneely, Julie P; Greer, Brett; Campbell, Katrina; Vasconcelos, Vitor; Elliott, Christopher T

    2014-05-01

    A highly sensitive broad specificity monoclonal antibody was produced and characterised for microcystin detection through the development of a rapid surface plasmon resonance (SPR) optical biosensor based immunoassay. The antibody displayed the following cross-reactivity: MC-LR 100%; MC-RR 108%; MC-YR 68%; MC-LA 69%; MC-LW 71%; MC-LF 68%; and Nodularin 94%. Microcystin-LR was covalently attached to a CM5 chip and with the monoclonal antibody was employed in a competitive 4 min injection assay to detect total microcystins in water samples below the WHO recommended limit (1 µg/L). A 'total microcystin' level was determined by measuring free and intracellular concentrations in cyanobacterial culture samples as this toxin is an endotoxin. Glass bead beating was used to lyse the cells as a rapid extraction procedure. This method was validated according to European Commission Decision 96/23/EC criteria. The method was proven to measure intracellular microcystin levels, the main source of the toxin, which often goes undetected by other analytical procedures and is advantageous in that it can be used for the monitoring of blooms to provide an early warning of toxicity. It was shown to be repeatable and reproducible, with recoveries from spiked samples ranging from 74 to 123%, and had % CVs below 10% for intra-assay analysis and 15% for inter-assay analysis. The detection capability of the assay was calculated as 0.5 ng/mL for extracellular toxins and 0.05 ng/mL for intracellular microcystins. A comparison of the SPR method with LC-MS/MS was achieved by testing six Microcystis aeruginosa cultures and this study yielded a correlation R(2) value of 0.9989. PMID:24720955

  15. Development of Multichannel Eddy Current Testing Instrument

    International Nuclear Information System (INIS)

    Four main techniques of electromagnetic testing are used for commercial applications: eddy current testing, alternating current field testing, magnetic flux leakage testing and remote field testing. Eddy current testing is a nondestructive evaluation method, which makes eddy current flow on a specimen by applying driving pulse to eddy current probe coil, by using eddy current testing device, and makes the change of eddy current which is dependently caused by flaws, material characteristics, testing condition, receiving through eddy current, and analyzes material properties, flaws, status on the specimen. Application of EC instrumentation varies widely in industry from the identification of metal heat treatment to the inspection of steam generator tubing in nuclear power plants. In this study, we have designed multichannel EC instrument which can be applicable to the NDE of the tube in heat exchanger for electric power facility, chemistry, and military industry, and finally confirmed the proper function of EC instrumentation

  16. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    Directory of Open Access Journals (Sweden)

    Anju Mohan

    2016-07-01

    Full Text Available Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT, microtiter plate agglutination test (MAT, indirect hemagglutination assay (IHA, and indirect enzyme-linked immunosorbent assay (iELISA as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001. The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005. The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002. However, the difference in mean iELISA titers of infected cattle (1.3678±0.014 and healthy vaccinated cattle (1.367±0.014 was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

  17. Comparison of five serological tests to detect Brucella abortus antibodies and a report on prevalence of the disease in livestock in the state of Yucatan, Mexico

    International Nuclear Information System (INIS)

    One hundred and seventy five sera samples from cattle were tested for reactions to 5 serological tests for Brucella antibodies. The tests used were Rose Bengal (RBT), 2-mercaptoethanol (2ME), rivanol (R), ELISA and complement fixation (CFT). The last of these was used as a standard against which the others were compared. Overall sensitivity and specificity for the 4 other tests by comparison were as follows: RBT, 100%, and 38%; 2ME, 90 and 99%; R, 86% and 100%, ELISA 97% and 99%. These figures varied within sub-groups of vaccinated and non-vaccinated animals. The ELISA test was able to differentiate vaccination and non-vaccination titres. Use of the RBT as a screening test followed by either CFT or ELISA for confirmation would yield the most reliable results. 2323 samples from cattle on 66 farms were tested with RBT as a screening test and 2ME for confirmation. 2.5% were positive for Brucella antigens. 1583 samples from pigs on 61 farms were tested by the same procedure and none was found to be positive for Brucella antibodies. (author). 13 refs, 1 fig., 5 tabs

  18. Development of a mouse monoclonal antibody against the chondroitin sulfate-protein linkage region derived from shark cartilage.

    Science.gov (United States)

    Akatsu, Chizuru; Fongmoon, Duriya; Mizumoto, Shuji; Jacquinet, Jean-Claude; Kongtawelert, Prachya; Yamada, Shuhei; Sugahara, Kazuyuki

    2010-05-01

    Glycosaminoglycans (GAGs) like chondroitin sulfate (CS) and heparan sulfate (HS) are synthesized on the tetrasaccharide linkage region, GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-O-Ser, of proteoglycans. The Xyl can be modified by 2-O-phosphate in both CS and HS, whereas the Gal residues can be sulfated at C-4 and/or C-6 in CS but not in HS. To study the roles of these modifications, monoclonal antibodies were developed against linkage glycopeptides of shark cartilage CS proteoglycans, and one was characterized in detail. This antibody bound hexa- and pentasaccharide-peptides more strongly than unsaturated tetrasaccharide-peptides with the unnatural fourth sugar residue (unsaturated hexuronic acid), suggesting the importance of the fifth and/or fourth saccharide residue GalNAc-5 and/or GlcA-4. Its reactivity was not affected by treatment with chondro-4-sulfatase or alkaline phosphatase, suggesting that 4-O-sulfate on the Gal residues and 2-O-phosphate on the Xyl residue were not recognized. Treatment with weak alkali to cleave the Xyl-Ser linkage completely abolished the binding activity, suggesting the importance of the peptide moiety of the hexasaccharide-peptide for the binding. Based on the amino acid composition and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses, it was revealed that the peptide moiety is composed of four amino acids, Ser, Pro, Gly, and Glu. Furthermore, the antibody stained wild-type CHO cells significantly, but much weakly mutant cells deficient in xylosyl- or galactosyltransferase-I required for the biosynthesis of the linkage region. These results suggest that the antibody recognizes the structure GalNAc(+/-6-O-sulfate)-GlcA-Gal-Gal-Xyl-Ser-(Pro, Gly, Glu). The antibody will be a useful tool for investigating the significance of the linkage region in the biosynthesis and/or intracellular transport of different GAG chains especially since such tools to study the linkage region are lacking. PMID:20336366

  19. Development of modified product consistency test

    International Nuclear Information System (INIS)

    Modified Product Consistency Test (M-PCT) has been developed as an alternative to other existing methods in determining the leachability of glass. M-PCT, the leaching method, is a hybrid of MCC-1 and PCT, but can provide quicker sample preparation. Larger diameter glass sample (1.0-2.0 mm) than in the PCT method can be used so that the glass beads are more easily produced and cleaned. From the M-PCT, the total mass loss (ML) of glass, the normalized elemental release rate (NLi), pH value of leachate have been obtained. For some selected glasses in which leaching rates have been known, their chemical durability have been using the M-PCT method. The results are compared to the literature data for the glasses. It is found that M-PCT method is reasonable and suitable in determining the leachability of Low and Intermediate Level Radioactive Waste glass form, such as the pH, elemental loss and total mass loss

  20. WAN emulation development and testing at Fermilab

    International Nuclear Information System (INIS)

    The Compact Muon Solenoid (CMS) experiment at CERN's Large Hadron Collider (LHC) is scheduled to come on-line in 2007. Fermilab will act as the CMS Tier-1 centre for the US and make experiment data available to more than 400 researchers in the US participating in the CMS experiment. The US CMS Users Facility group, based at Fermilab, has initiated a project to develop a model for optimizing movement of CMS experiment data between CERN and the various tiers of US CMS data centres and to design a WAN emulation facility which will enable controlled testing of unmodified or modified CMS applications and TCP implementations locally under conditions that emulate WAN connectivity. The WAN emulator facility is configurable for latency, jitter, and packet loss. The initial implementation is based on the NISTnet software product. In this paper we will describe the status of this project to date, the results of validation and comparison of performance measurements obtained in emulated and real environment for different applications including multistreams GridFTP. We also will introduce future short term and intermediate term plans, as well as outstanding problems and issues

  1. Development of robust antibody purification by optimizing protein-A chromatography in combination with precipitation methodologies.

    Science.gov (United States)

    Chollangi, Srinivas; Parker, Ray; Singh, Nripen; Li, Yi; Borys, Michael; Li, Zhengjian

    2015-11-01

    To be administered to patients, therapeutic monoclonal antibodies must have very high purity, with process related impurities like host-cell proteins (HCPs) and DNA reduced to Precipitation with low pH treatment of cell culture harvest resulted in selective removal of impurities while manipulating the pH of wash buffers used in Protein-A chromatography and incorporating wash additives that disrupt various modes of protein-protein interaction resulted in further and more pronounced reduction in impurity levels. In addition, our study also demonstrate that optimizing the neutralization pH post Protein-A elution can result in selective removal of impurities. When applied over multiple mAbs, this optimization method proved to be very robust and the strategy provides a new and improved purification process that reduces process related impurities like HCPs and DNA to drug substance specifications with just one chromatography column and open avenues for significant decrease in operating costs in monoclonal antibody purification. PMID:25950654

  2. Development and evaluation of the rVP-ELISA for detection of antibodies against porcine parvovirus.

    Science.gov (United States)

    Kong, Miaomiao; Peng, Yonggang; Cui, Yuchao; Chang, Tiecheng; Wang, Xiaoling; Liu, Zhaoxia; Liu, Yonggang; Zhu, Yu; Luo, Yakun; Tang, Qinghai; Feng, Li; Cui, Shangjin

    2014-09-01

    The gene encoding the VP2 protein of porcine parvovirus (PPV) was expressed in an insect-baculovirus system. The recombinant (r) VP2 was similar antigenically/functionally to the native capsid protein as demonstrated by hemagglutination (HA), Western blotting using PPV positive sera. The purified rVP2 proteins were used as coating antigen to establish a rVP-ELISA method for detection of PPV positive and negative sera from pigs. The optimal operating conditions of the rVP-ELISA were: the concentration of rVP2 proteins coated on the wells was 2 μg/mL; the diluted concentration of serum was 1: 150 and that of the enzyme-labeled antibody was 1: 6000. A total of 596 sera were detected by this assay, and the average positive rate was 87%. Compared with France LSI kit, the result showed that the coincidence rate was 96.7%. In conclusion, the rVP2-ELISA is a sensitive and specific method for detecting antibodies against PPV. PMID:24945904

  3. Antithyroid microsomal antibody

    Science.gov (United States)

    Thyroid antimicrosomal antibody; Antimicrosomal antibody; Microsomal antibody; Thyroid peroxidase antibody; TPOAb ... Granulomatous thyroiditis Hashimoto thyroiditis High levels of these antibodies have also been linked to an increased risk ...

  4. Clinical Significance of HLA-DQ Antibodies in the Development of Chronic Antibody-Mediated Rejection and Allograft Failure in Kidney Transplant Recipients.

    Science.gov (United States)

    Lee, Hyeyoung; Min, Ji Won; Kim, Ji-Il; Moon, In-Sung; Park, Ki-Hyun; Yang, Chul Woo; Chung, Byung Ha; Oh, Eun-Jee

    2016-03-01

    With the development of the single antigen beads assay, the role of donor specific alloantibody (DSA) against human leukocyte antigens in kidney transplantation (KT) has been highlighted. This study aimed to investigate the clinical significance of DQ-DSA detected at renal allograft biopsy. We evaluated 263 KT recipients who underwent allograft biopsy and DSA detection at the same time. Among them, 155 patients who were nonsensitized before transplantation were selected to investigate the role of de-novo DQ-DSA. Both the total and nonsensitized subgroup was categorized into 4 groups each according to DSA results as: DQ only, DQ + non-DQ, non-DQ, and no DSA. In the total patient group, post-KT DSA was positive in 79 (30.0%) patients and DQ-DSA was most prevalent (64.6%). In the nonsensitized subgroup, de-novo DSAs were detected in 45 (29.0%) patients and DQ-DSA was also most prevalent (73.3%). The DQ only group showed a significantly longer post-KT duration compared to the other groups (P < 0.05). The overall incidence of antibody-mediated rejection (AMR) was 17.9%. B-DSA, DR-DSA, and DQ-DSA were associated with AMR (P < 0.05), but in the analysis for chronic AMR, only DQ-DSA showed significance in both the total and the nonsensitized subgroup (P < 0.05). On comparison of Banff scores among groups, those representing humoral immunity were significantly dominant in all DSA positive groups compared to the no DSA group (P < 0.05), and higher scores of markers representing chronic tissue injury were more frequently detected in the groups with DQ-DSA. The worst postbiopsy survival was seen in the DQ + non-DQ group of the total patient group, and patients with de-novo DQ-DSA showed poorer graft survival in the nonsensitized subgroup compared to the no DSA group (P < 0.05). In the multivariate analysis, de-novo DQ-DSA was the only significant risk factor associated with late allograft failure (P < 0.05). Our study is the first to demonstrate

  5. Production of Monoclonal Antibodies Against the Challenge Strain of Infectious Laryngotracheitis Virus of Chickens and Their Use in an Indirect Immunofluorescent Diagnostic Test

    Directory of Open Access Journals (Sweden)

    Ferhat Abbas*, James Andreasen1, Rockey Becker1, Masroor Ahmed, M Arif Awan, Abdul Wadood and Anita Sonn1

    2010-07-01

    Full Text Available The objective of the present research was to produce monoclonal antibodies (MCAs against the USDA challenge strain of infectious laryngotracheitis virus and to perform an initial investigation of their use in an indirect immunofluorescence diagnostic test. Fourteen-day old chicken embryo liver cells were grown in tissue culture plates. Confluent monolayers were obtained after 48 hours. Monolayers were infected with the USDA challenge strain of infectious laryngotracheitis virus (ILTV. Cytopethic effect of the virus in the form of syncytial formation and clumping of cells was observed after 24 hours. The virus from the tissue culture flasks was collected and purified using discontinuous sucrose gradient. A clear band of the virus from sucrose gradient was obtained. The refractory index and the density measured were 1.410 and 1.20 g/cm3, respectively. Spectrophotometry of the purified virus showed 68.117 ug/ml of protein and 9.8948 ug/ml of nucleic acid concentration. Spleen cells from immunized mice with pure virus were fused with myeloma cells and hybridomas were obtained after 10 days. Screening was performed using indirect immunofluorescence antibody test (IFAT using rabbit anti-mouse immunoglobulins as secondary antibodies. Three hybridomas, 2D1D8, 2E11G2 and 2C6C7 were found producing antibodies against ILTV. All monoclonal antibodies were of isotype IgM and reacted with different strains of ILTV (ILTV USDA, S 88 00224, 86-1169 in IFAT. None of the monoclonals reacted with Parrot herpesvirus and avian adenovirus 301 in IFAT.

  6. An Investigation into the Development of a Spoken English Test

    Institute of Scientific and Technical Information of China (English)

    付期棉

    2014-01-01

    This paper investigates the development of a spoken English test. The nature of speaking test and its design principles are first reviewed. Then the procedure of the test development is elaborated in detail,namely,design stage,construction stage and try out stage. The challenges facing the development of spoken test are finally discussed.

  7. Clinical Comparison of the Treponema pallidum CAPTIA Syphilis-G Enzyme Immunoassay with the Fluorescent Treponemal Antibody Absorption Immunoglobulin G Assay for Syphilis Testing

    OpenAIRE

    Halling, V. W.; Jones, M. F.; Bestrom, J. E.; Wold, A D; Rosenblatt, J E; Smith, T. F.; Cockerill, F R

    1999-01-01

    Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphilis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results of rapid plasmid reagin (RPR) testing of the same specimens. Borderline CAPTI...

  8. Clinical appraisal of patients and detection of serum antibodies by ELISA and CIA tests in late periods of Trichinella sp. invasion

    Directory of Open Access Journals (Sweden)

    Kociecka W.

    2001-06-01

    Full Text Available Results of our studies using ELISA and competitive inhibition assay (CIA tests fully confirmed the previously experienced trichinellosis and reflected persistent stimulation of antibody production due to the antigen release from Trichinella larvae, which had survived longer and undergone gradual destruction in the muscles. The studies proved that the tests complement each other, yielding concordant results in 86.7 % of cases. Due to its higher specificity, the CIA test can help in interpreting pathological signs/symptoms and in evaluating humoral response activity at late and distant in time periods following the invasion.

  9. Production of Monoclonal Antibodies Against the Challenge Strain of Infectious Laryngotracheitis Virus of Chickens and Their Use in an Indirect Immunofluorescent Diagnostic Test

    OpenAIRE

    Ferhat Abbas*, James Andreasen1, Rockey Becker1, Masroor Ahmed, M Arif Awan, Abdul Wadood and Anita Sonn1

    2010-01-01

    The objective of the present research was to produce monoclonal antibodies (MCAs) against the USDA challenge strain of infectious laryngotracheitis virus and to perform an initial investigation of their use in an indirect immunofluorescence diagnostic test. Fourteen-day old chicken embryo liver cells were grown in tissue culture plates. Confluent monolayers were obtained after 48 hours. Monolayers were infected with the USDA challenge strain of infectious laryngotracheitis virus (ILTV). Cytop...

  10. Evaluation of new monoclonal antibody-based latex agglutination test for detection of cryptococcal polysaccharide antigen in serum and cerebrospinal fluid.

    OpenAIRE

    Kiska, D L; Orkiszewski, D R; Howell, D; Gilligan, P H

    1994-01-01

    We evaluated the performance of CRYPTO-LEX (Trinity Laboratories, Inc., Raleigh, N. C.), a new mouse immunoglobulin M monoclonal antibody latex agglutination reagent which reacts with the capsular polysaccharide of the four serogroups of Cryptococcus neoformans. This test was compared with CALAS (Meridian Diagnostics, Cincinnati, Ohio) for the ability to detect cryptococcal antigen in serum and cerebrospinal fluid (CSF). A total of 580 clinical specimens (327 serum and 253 CSF samples), prima...

  11. Humoral Immune Response Kinetics in Philander opossum and Didelphis marsupialis Infected and Immunized by Trypanosoma cruzi Employing an Immunofluorescence Antibody Test

    Directory of Open Access Journals (Sweden)

    Ana Paula Legey

    1999-05-01

    Full Text Available Philander opossum and Didelphis marsupialis considered the most ancient mammals and an evolutionary success, maintain parasitism by Trypanosoma cruzi without developing any apparent disease or important tissue lesion. In order to elucidate this well-balanced interaction, we decided to compare the humoral immune response kinetics of the two didelphids naturally and experimentally infected with T. cruzi and immunized by different schedules of parasite antigens, employing an indirect fluorescence antibody test (IFAT. Both didelphids responded with high serological titers to different immunization routes, while the earliest response occurred with the intradermic route. Serological titers of naturally infected P. opossum showed a significant individual variation, while those of D. marsupialis remained stable during the entire follow-up period. The serological titers of the experimentally infected animals varied according to the inoculated strain. Our data suggest that (1 IFAT was sensitive for follow-up of P. opossum in natural and experimental T. cruzi infections; (2 both P. opossum and D. marsupialis are able to mount an efficient humoral immune response as compared to placental mammals; (3 experimentally infected P. opossum and D. marsupialis present distinct patterns of infection, depending on the subpopulation of T. cruzi, (4 the differences observed in the humoral immune responses between P. opossum and D. marsupialis, probably, reflect distinct strategies selected by these animals during their coevolution with T. cruzi.

  12. Humoral immune response kinetics in Philander opossum and Didelphis marsupialis infected and immunized by Trypanosoma cruzi employing an immunofluorescence antibody test.

    Science.gov (United States)

    Legey, A P; Pinho, A P; Chagas Xavier, S C; Leon, L L; Jansen, A M

    1999-01-01

    Philander opossum and Didelphis marsupialis considered the most ancient mammals and an evolutionary success, maintain parasitism by Trypanosoma cruzi without developing any apparent disease or important tissue lesion. In order to elucidate this well-balanced interaction, we decided to compare the humoral immune response kinetics of the two didelphids naturally and experimentally infected with T. cruzi and immunized by different schedules of parasite antigens, employing an indirect fluorescence antibody test (IFAT). Both didelphids responded with high serological titers to different immunization routes, while the earliest response occurred with the intradermic route. Serological titers of naturally infected P. opossum showed a significant individual variation, while those of D. marsupialis remained stable during the entire follow-up period. The serological titers of the experimentally infected animals varied according to the inoculated strain. Our data suggest that (1) IFAT was sensitive for follow-up of P. opossum in natural and experimental T. cruzi infections; (2) both P. opossum and D. marsupialis are able to mount an efficient humoral immune response as compared to placental mammals; (3) experimentally infected P. opossum and D. marsupialis present distinct patterns of infection, depending on the subpopulation of T. cruzi, (4) the differences observed in the humoral immune responses between P. opossum and D. marsupialis, probably, reflect distinct strategies selected by these animals during their coevolution with T. cruzi. PMID:10348985

  13. Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of Xanthomonas hyacinthi

    Energy Technology Data Exchange (ETDEWEB)

    Doorn, J. van; Ojanen-Reuhs, T.; Hollinger, T.C.; Reuhs, B.L.; Schots, A.; Boonekamp, P.M.; Oudega, B.

    1999-09-01

    The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy; however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths.

  14. Developing a Noninvasive Procedure Using Labeled Monoclonal Antibody Anti-VEGF (Bevacizumab for Detection of Endometriosis

    Directory of Open Access Journals (Sweden)

    Daniel Escorsim Machado

    2015-01-01

    Full Text Available The off-label use of bevacizumab labeled with 99mTc as a new radiopharmaceutical for imaging of endometriosis is a promising noninvasive, new clinical procedure. The bevacizumab in monoclonal antibodies targeted at vascular endothelial growth factor (VEGF is superexpressed in cases of endometriosis. In this study we evaluate the imaging of endometriosis lesion in rats (induced to endometriosis using bevacizumab-99mTc. The results showed that bevacizumab-99mTc imaged the lesion and support his use for Nuclear Medicine applied to gynecology. Also the results appointed that this radiopharmaceutical has a hepatobiliary excretion. It is important to notice that the dose used was almost 0,01% of the usual dose for the bevacizumab.

  15. Development of antibodies to protective antigen and lethal factor components of anthrax toxin in humans and guinea pigs and their relevance to protective immunity.

    OpenAIRE

    Turnbull, P. C.; Broster, M G; Carman, J A; Manchee, R J; Melling, J

    1986-01-01

    A competitive inhibition enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies in serum to the protective antigen (PA) and lethal factor (LF) components of anthrax toxin. Current human vaccination schedules with an acellular vaccine induce predictable and lasting antibody titers to PA and, when present in the vaccine, to LF. Live spore vaccine administered to guinea pigs in a single dose conferred significantly better protection than the human vaccines (P less than 0.00...

  16. SLAC physicists develop test for string theory

    CERN Multimedia

    Yajnik, Juhi

    2006-01-01

    "Under certain conditions, string theory solves many of the questions wracking the minds of physicists, but until recently it had one major flaw - it could not be tested. SLAC (Stanford Linear Accelerator Center) scientists have found a way to test this revolutionary theory, which posits that there are 10 or 11 dimensions in our universe" (1 page)

  17. Development of a specific monoclonal antibody-based ELISA to measure the artemether content of antimalarial drugs.

    Directory of Open Access Journals (Sweden)

    Suqin Guo

    Full Text Available Artemether is one of the artemisinin derivatives that are active ingredients in antimalarial drugs. Counterfeit and substandard antimalarial drugs have become a serious problem, which demands reliable analytical tools and implementation of strict regulation of drug quality. Structural similarity among artemisinin analogs is a challenge to develop immunoassays that are specific to artemisinin derivatives. To produce specific antibodies to artemether, we used microbial fermentation of artemether to obtain 9-hydroxyartemether, which was subsequently used to prepare a 9-O-succinylartemether hapten for conjugation with ovalbumin as the immunogen. A monoclonal antibody (mAb, designated as 2G12E1, was produced with high specificity to artemether. 2G12E1 showed low cross reactivities to dihydroartemisinin, artemisinin, artesunate and other major antimalarial drugs. An indirect competitive enzyme linked immunosorbent assay (icELISA developed showed a concentration causing 50% of inhibition for artemether as 3.7 ng mL⁻¹ and a working range of 0.7-19 ng mL⁻¹. The icELISA was applied for determination of artemether content in different commercial drugs and the results were comparable to those determined by high-performance liquid chromatography analysis. In comparison with reported broad cross activity of anti-artemisinin mAbs, the most notable advantage of the 2G12E1-based ELISA is its high specificity to artemether only.

  18. Development of a Specific Monoclonal Antibody for the Quantification of Artemisinin in Artemisia annua and Rat Serum.

    Science.gov (United States)

    Guo, Suqin; Cui, Yongliang; Wang, Kunbi; Zhang, Wei; Tan, Guiyu; Wang, Baomin; Cui, Liwang

    2016-03-01

    Artemisinin, extracted from Artemisia annua, and its derivatives are important frontline antimalarials. To produce specific antibodies for the detection and quantification of artemisinin, artemisinin was transformed to 9-hydroxyartemisinin by microbial fermentation, which was used to prepare a 9-succinate artemisinin hapten for conjugation with ovalbumin. A monoclonal antibody (mAb), designated as 3H7A10, was selected from hybridoma cell lines which showed high specificity to artemisinin. No competitive inhibition was observed with artesunate, dihydroartemisinin, and artemether for up to 20,000 ng mL(-1). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed, which showed a concentration causing 50% of inhibition (IC50) for artemisinin as 2.6 ng mL(-1) and a working range of 0.6-11.5 ng mL(-1). The icELISA was applied for the quantification of artemisinin in crude extracts of wild A. annua and the study of pharmacokinetics of artemisinin in rat serum after intraperitoneal injection. The results were highly correlated with those determined by HPLC-UV analysis (R(2) = 0.9919). In comparison with reported antiartemisinin mAbs which have broad cross-reactivity with other artemisinin derivatives, the high specificity of 3H7A10 for artemisinin will enable development of methods for quantification of artemisinin in Artemisia plants and antimalarial drugs such as Arco and for pharmacokinetic studies. PMID:26822789

  19. Application Value of Irregular Antibody Test Before Transfusion%输血前不规则抗体检验的应用价值分析

    Institute of Scientific and Technical Information of China (English)

    连海燕; 李正花

    2015-01-01

    目的:分析输血前不规则抗体检验的临床价值,为临床安全输血提供有力指导。方法整群选取需输血治疗患者2278例,输血前均展开不规则抗体筛查。结果2278例输血患者共16例不规则抗体检验阳性,阳性率为0.702%;女性患者不规则抗体阳性率明显高于男性,孕妇不规则抗体阳性率明显高于非孕妇。16例不规则抗体阳性患者抗体类型为:非特异性抗体1例,抗-C2例,抗-E3例,抗-D4例,抗-M6例。结论在输血前为患者展开不规则抗体检验,有助于提高输血安全性,降低输血不良反应发生风险,值得推广。%Objective To analyze the clinical value of irregular antibody test before blood transfusion and provide guidance for transfusion safety. Methods 2278 patients who needed blood transfusion underwent irregular antibody test before blood transfusion. Results Out of 2278 patients, 16 patients (0.702%) were found with positive irregular antibody in which there were more female than male, more pregnant women than non-pregnancy women, and the antibody types included 1 case of non-specific antibodies, 2 cases of anti-C, 3 cases of anti-E, 4 cases of anti-D, 6 cases of anti-M. Conclusion Irregular antibody test before blood trans-fusion can improve transfusion safety and reduce the risk of adverse reaction, therefore it is worthy of promotion.

  20. A pilot study comparing the development of EIAV Env-specific antibodies induced by DNA/recombinant vaccinia-vectored vaccines and an attenuated Chinese EIAV vaccine.

    Science.gov (United States)

    Meng, Qinglai; Lin, Yuezhi; Ma, Jian; Ma, Yan; Zhao, Liping; Li, Shenwei; Yang, Kai; Zhou, Jianhua; Shen, Rongxian; Zhang, Xiaoyan; Shao, Yiming

    2012-12-01

    Data from successful attenuated lentiviral vaccine studies indicate that fully mature Env-specific antibodies characterized by high titer, high avidity, and the predominant recognition of conformational epitopes are associated with protective efficacy. Although vaccination with a DNA prime/recombinant vaccinia-vectored vaccine boost strategy has been found to be effective in some trials with non-human primate/simian/human immunodeficiency virus (SHIV) models, it remains unclear whether this vaccination strategy could elicit mature equine infectious anemia virus (EIAV) Env-specific antibodies, thus protecting vaccinated horses against EIAV infection. Therefore, in this pilot study we vaccinated horses using a strategy based on DNA prime/recombinant Tiantan vaccinia (rTTV)-vectored vaccines encoding EIAV env and gag genes, and observed the development of Env-specific antibodies, neutralizing antibodies, and p26-specific antibodies. Vaccination with DNA induced low titer, low avidity, and the predominant recognition of linear epitopes by Env-specific antibodies, which was enhanced by boosting vaccinations with rTTV vaccines. However, the maturation levels of Env-specific antibodies induced by the DNA/rTTV vaccines were significantly lower than those induced by the attenuated vaccine EIAV(FDDV). Additionally, DNA/rTTV vaccines did not elicit broadly neutralizing antibodies. After challenge with a virulent EIAV strain, all of the vaccinees and control horses died from EIAV disease. These data indicate that the regimen of DNA prime/rTTV vaccine boost did not induce mature Env-specific antibodies, which might have contributed to immune protection failure. PMID:23171359

  1. Texas Hospitals Develop Rapid Test for Zika Virus Detection

    Science.gov (United States)

    ... 157415.html Texas Hospitals Develop Rapid Test for Zika Virus Detection But it will only be available at ... News) -- The first rapid detection test for the Zika virus has been developed by teams at two Texas ...

  2. Dot-ELISA affinity test: an easy, low-cost method to estimate binding activity of monoclonal antibodies

    Czech Academy of Sciences Publication Activity Database

    Svobodová, Z.; Jankovičová, B.; Horák, Daniel; Bílková, Z.

    2013-01-01

    Roč. 4, č. 3 (2013), 1000168_1-1000168_5. ISSN 2155-9872 R&D Projects: GA MŠk 7E12053; GA MŠk 7E09109 EU Projects: European Commission(XE) 246513 - NADINE; European Commission(XE) 228980 - CAMINEMS Institutional support: RVO:61389013 Keywords : dot-ELISA * affinity * monoclonal antibody Subject RIV: CD - Macromolecular Chemistry

  3. TSH receptor antibodies in subjects with type 1 diabetes mellitus.

    Science.gov (United States)

    Unnikrishnan, Ambika G; Kumaravel, Velayutham; Nair, Vasantha; Rao, Ananth; Jayakumar, Rohini V; Kumar, Harish; Sanjeevi, Carani B

    2006-10-01

    The research was undertaken to study the prevalence of TSH receptor antibody positivity in patients with type 1 diabetes. A total of 74 subjects with type 1 diabetes were enrolled in this cross-sectional study. Thyroid function test and assessment of thyroid autoimmunity with anti-TPO and TSH receptor antibody were done in all patients. A total of 33 males and 41 females with type 1 diabetes were studied. The prevalence of TSH receptor antibody positivity alone was 18%. The prevalence of thyroid autoimmunity with anti-TPO as a marker was 28%; the prevalence increased to 43% when TSH receptor antibody was also measured. Majority of the subjects with antithyroid antibody positivity were also positive for GAD65 antibodies. As a significant proportion of type 1 diabetic subjects have positivity to TSH receptor antibody, we suggest that larger studies should be conducted to study the benefits of TSH receptor antibody-based screening for thyroid dysfunction in type 1 diabetic subjects. As the TSH receptor antibodies could be of the stimulating or of the blocking type, subjects with antibody positivity could be at risk of developing hyperthyroidism or hypothyroidism. PMID:17130558

  4. Penetration Testing in Agile Software Development Projects

    OpenAIRE

    Tomanek, Martin; Klima, Tomas

    2015-01-01

    Agile development methods are commonly used to iteratively develop the information systems and they can easily handle ever-changing business requirements. Scrum is one of the most popular agile software development frameworks. The popularity is caused by the simplified process framework and its focus on teamwork. The objective of Scrum is to deliver working software and demonstrate it to the customer faster and more frequent during the software development project. However the security requir...

  5. Test driven development with Symfony2

    OpenAIRE

    Škrlep, Matej

    2014-01-01

    With the advance of technologies used in the software development process, the applications become more complex. Classic methodologies of software development are becoming a big hurdle because of their rigidity. As a consequence, agile software development methodologies are becoming more popular. They demand more communication between the customer and the development team. As a result, both parties better understand the application which leads to better customer satisfaction. With short deve...

  6. Deep sequencing and human antibody repertoire analysis.

    Science.gov (United States)

    Boyd, Scott D; Crowe, James E

    2016-06-01

    In the past decade, high-throughput DNA sequencing (HTS) methods and improved approaches for isolating antigen-specific B cells and their antibody genes have been applied in many areas of human immunology. This work has greatly increased our understanding of human antibody repertoires and the specific clones responsible for protective immunity or immune-mediated pathogenesis. Although the principles underlying selection of individual B cell clones in the intact immune system are still under investigation, the combination of more powerful genetic tracking of antibody lineage development and functional testing of the encoded proteins promises to transform therapeutic antibody discovery and optimization. Here, we highlight recent advances in this fast-moving field. PMID:27065089

  7. Development and evaluation of diagnostic tests for the serological diagnosis of brucellosis in swine

    Directory of Open Access Journals (Sweden)

    Tiziana Di Febo

    2012-06-01

    Full Text Available A competitive enzyme-linked immunosorbent assay (c-ELISA, an indirect ELISA (i-ELISA and a dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA were developed to test for antibodies to Brucella suis in pig and wild boar sera. An anti-Brucella-LPS monoclonal antibody (MAb 4B5A (c-ELISA and DELFIA and an anti-swine IgG monoclonal antibody (MAb 10C2G5 (i-ELISA were used for the three assays. The specificity (Sp and sensitivity (Se of the assays gave the following results: Se and Sp = 100% at a cut-off value of 61.0% (B/B0% for c-ELISA; Sp = 99.1% and Se = 100% at a cut-off value of 21.7% (percentage positivity: PP% for i-ELISA; Sp = 91.0% and Se = 75% at a cut-off value of 37.0% (B/B0% for DELFIA. In addition, the performance of a commercial fluorescence polarisation assay (FPA, standardised for bovine sera, was evaluated in swine sera. The specificity and sensitivity obtained were both 100% at a cut-off value of 99.5 (millipolarisation unit values. These results suggest that the combination of c-ELISA, i-ELISA and FPA can be used to improve the serological diagnosis of swine brucellosis.

  8. Comparison between sensitivity of autologous skin serum test and autologous plasma skin test in patients with Chronic Idiopathic Urticaria for detection of antibody against IgE or IgE receptor (FcεRIα).

    Science.gov (United States)

    Sajedi, Vahid; Movahedi, Masoud; Aghamohammadi, Asghar; Aghamohamadi, Asghar; Gharagozlou, Mohammad; Ghareguzlou, Mohammad; Shafiei, Alireza; Soheili, Habib; Sanajian, Nahal

    2011-06-01

    Intradermal injection of autologous serum and plasma elicit a cutaneous reactivity in almost 45-60% of patients with Chronic Idiopathic Urticaria (CIU). This reactivity is associated with the presence of auto antibodies against IgE or IgE receptors. This study was carried out to compare the cutaneous reactivity of autologous serum and plasma skin tests in a series of patients with CIU for diagnosis of auto antibodies against IgE or IgE receptor. Fifty eight patients with CIU were injected intradermally with autologous serum and plasma (anticoagulated by citrate). Histamine was used as positive control and normal saline as negative control. The study group was checked by routine laboratory tests (CBC, U/A etc), allergens with skin prick tests, and serum IgE level, and auto antibodies against thyroid as well. Duration of urticaria was another factor which was assessed.There was no significant difference between positive ASST and positive APST patients for the above mentioned tests. 77.6% of the patients were Positive for APST and 65.5% were ASST positive. Duration of urticaria was longer in patients with positive ASST and APST than ASST and APST negative patients, although the difference was not statistically significant.Autologus serum skin test (ASST) and autologous plasma skin test (APST) could be used for estimation of duration and severity of urticaria and planning for the treatment. PMID:21625019

  9. Development and Comparative Evaluation of a Competitive ELISA with Rose Bengal Test and a Commercial Indirect ELISA for Serological Diagnosis of Brucellosis

    OpenAIRE

    Mythili, Tadepalli; Rajendra, Lingala; Bhavesh, Trangadia; Thiagarajan, Dorairajan; Srinivasan, Villuppanoor Alwar

    2011-01-01

    The development of a competitive ELISA for the detection of brucella-specific antibodies in bovines is described. Anti-brucella guinea pig serum was used as a source of competing antibodies. Lipo-polysaccharide purified from inactivated B. abortus S19 culture was used as antigen for the development of the assay. Sera from cattle were used in the competitive ELISA, rose bengal test and a commercial indirect ELISA. The following cattle sera were tested: (i) known positive sera (n = 80) (ii) kno...

  10. Development of a multiplexed fluorescent immunoassay for the quantitation of antibody responses to four Neisseria meningitidis serogroups.

    Science.gov (United States)

    Martins, Thomas B; Jaskowski, Troy D; Tebo, Anne; Hill, Harry R

    2009-03-15

    Neisseria meningitidis is a gram-negative bacterium causing disease world wide with a fatality rate of 5-10%. Five serogroups, A, B, C, Y and W-135 are responsible for virtually all cases of the disease in humans. We have developed a multiplexed assay for the simultaneous quantitation of IgG antibody responses to the four most immunogenic (A, C, Y, and W-135) N. meningitidis serogroups. A simple and less manipulative method was employed for conjugation of the capsular polysaccharide antigens to the microspheres. The multiplex assay compared well with traditional individual ELISAs, but demonstrated greater than 1 log increase in dynamic range and sensitivity. Specificity studies of the multiplex assay showed greater than 95% homologous inhibition and less than 5% heterologous inhibition for all four serogroups. Intra and inter-assay CVs were generally less than 10% and the limit of detection was <600 pg/ml. The multiplexed assay proved to be reproducible as well as specific and sensitive when compared to the standardized ELISAs. Advantages included a greater dynamic range and simultaneous detection of antibody responses to the four serogroups contained in the tetravalent meningococcal polysaccharide vaccine. PMID:19159627

  11. Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies.

    Science.gov (United States)

    Tamošiūnas, Paulius Lukas; Petraitytė-Burneikienė, Rasa; Lasickienė, Rita; Akatov, Artiomas; Kundrotas, Gabrielis; Sereika, Vilimas; Lelešius, Raimundas; Žvirblienė, Aurelija; Sasnauskas, Kęstutis

    2014-01-01

    Porcine parvovirus (PPV) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the major capsid protein VP2 of PPV was amplified using viral nucleic acid extract from swine serum and inserted into yeast Saccharomyces cerevisiae expression plasmid. Recombinant PPV VP2 protein was efficiently expressed in yeast and purified using density gradient centrifugation. Electron microscopy analysis of purified PPV VP2 protein revealed the self-assembly of virus-like particles (VLPs). Nine monoclonal antibodies (MAbs) against the recombinant PPV VP2 protein were generated. The specificity of the newly generated MAbs was proven by immunofluorescence analysis of PPV-infected cells. Indirect IgG ELISA based on the recombinant VLPs for detection of PPV-specific antibodies in swine sera was developed and evaluated. The sensitivity and specificity of the new assay were found to be 93.4% and 97.4%, respectively. In conclusion, yeast S. cerevisiae represents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance. PMID:25045718

  12. Generation of Recombinant Porcine Parvovirus Virus-Like Particles in Saccharomyces cerevisiae and Development of Virus-Specific Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Paulius Lukas Tamošiūnas

    2014-01-01

    Full Text Available Porcine parvovirus (PPV is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. The gene coding for the major capsid protein VP2 of PPV was amplified using viral nucleic acid extract from swine serum and inserted into yeast Saccharomyces cerevisiae expression plasmid. Recombinant PPV VP2 protein was efficiently expressed in yeast and purified using density gradient centrifugation. Electron microscopy analysis of purified PPV VP2 protein revealed the self-assembly of virus-like particles (VLPs. Nine monoclonal antibodies (MAbs against the recombinant PPV VP2 protein were generated. The specificity of the newly generated MAbs was proven by immunofluorescence analysis of PPV-infected cells. Indirect IgG ELISA based on the recombinant VLPs for detection of PPV-specific antibodies in swine sera was developed and evaluated. The sensitivity and specificity of the new assay were found to be 93.4% and 97.4%, respectively. In conclusion, yeast S. cerevisiae represents a promising expression system for generating recombinant PPV VP2 protein VLPs of diagnostic relevance.

  13. Correlation between the presence of neutralizing antibodies against porcine circovirus 2 (PCV2 and protection against replication of the virus and development of PCV2-associated disease

    Directory of Open Access Journals (Sweden)

    Bøtner Anette

    2006-01-01

    Full Text Available Abstract Background In a previous study, it was demonstrated that high replication of Porcine circovirus 2 (PCV2 in a gnotobiotic pig was correlated with the absence of PCV2-neutralizing antibodies. The aim of the present study was to investigate if this correlation could also be found in SPF pigs in which PMWS was experimentally reproduced and in naturally PMWS-affected pigs. Results When looking at the total anti-PCV2 antibody titres, PMWS-affected and healthy animals seroconverted at the same time point, and titres in PMWS-affected animals were only slightly lower compared to those in healthy animals. In healthy animals, the evolution of PCV2-neutralizing antibodies coincided with that of total antibodies. In PMWS-affected animals, neutralizing antibodies could either not be found (sera from field studies or were detected in low titres between 7 and 14 DPI only (sera from experimentally inoculated SPF pigs. Differences were also found in the evolution of specific antibody isotypes titres against PCV2. In healthy pigs, IgM antibodies persisted until the end of the study, whereas in PMWS-affected pigs they quickly decreased or remained present at low titres. The mean titres of other antibody isotypes (IgG1, IgG2 and IgA, were slightly lower in PMWS-affected pigs compared to their healthy group mates at the end of each study. Conclusion This study describes important differences in the development of the humoral immune response between pigs that get subclinically infected with PCV2 and pigs that experience a high level of PCV2-replication which in 3 of 4 experiments led to the development of PMWS. These observations may contribute to a better understanding of the pathogenesis of a PCV2-infection.

  14. Development of [{sup 201}Tl](III)-DTPA-human polyclonal antibody complex for inflammation detection

    Energy Technology Data Exchange (ETDEWEB)

    Jalilian, A.R.; Kamali-Dehghan, M.; Kamrani, Y.Y. [Nuclear Research Center for Agriculture and Medicine, Karaj (Iran). Cyclotron and Nuclear Medicine Dept.; Khorrami, A.; Tavakoli, M.B. [Medical Sciences Univ. of Isfahan (Iran). Medical Physics and Engineering Dept.

    2007-07-01

    Thallium-201 (T{sub 1/2}=3.04 d) in Tl{sup +} form was converted to Tl{sup 3+} cation in presence of O{sub 3} in 6 M HCl controlled by RTLC/gel electrophoresis methods and used in the labeling of human polyclonal antibody (HIgG) after conjugation with freshly prepared cyclic DTPA-dianhydride. The best results of the conjugation were obtained by the addition of 1 mL of a HIgG pharmaceutical solution (5 mg/ml, in phosphate buffer, pH=7) to a glass tube pre-coated with DTPA-dianhydride (0.01 mg) at 25 C with continuous mild stirring for 30 min. The final isotonic [{sup 201}Tl](III)-DTPA-HIgG complex was checked by radio-TLC using several solvent systems to ensure the formation of only one species followed by filtration through a 0.22 {mu} filter (specific activity= 33.7 TBq/mM, radiochemical purity >95%). Preliminary bio-distribution studies in normal and inflammation-bearing rats were performed. The target/skin and target/blood ratios were 4 and 6 after 28 h respectively, showing the selectivity of the radiopharmaceutical for the inflammatory lesions. (orig.)

  15. Measurement of tumour reactive antibody and antibody conjugate by competition, quantitated by flow cytofluorimetry.

    Science.gov (United States)

    Robins, R A; Laxton, R R; Garnett, M; Price, M R; Baldwin, R W

    1986-06-24

    Binding of unlabelled monoclonal antibody preparations has been assessed by competition at saturation with fluorochrome labelled homologous antibody for binding to antigen bearing target cells. The extent of competition was measured by quantitative flow cytofluorimetry, and simple mathematical procedures have been developed to allow the interpretation of competition data in terms of antibody binding activity. In the system studied, non-specific (non-competitive) fluorescence was minimal, but an iterative method to calculate its contribution to the measured signal is given. This approach has the advantage that the antibody preparation to be tested does not need to be labelled or modified; this is particularly important when evaluating the binding activity of therapeutic antibody conjugates. Comparison with a well characterized standard antibody preparation provides a rapid, sensitive and accurate quality control procedure. This test is also simple to perform, requiring only the mixing of labelled and unlabelled antibodies with target cells, a single incubation, followed by analysis without washing of the target cells. PMID:2424997

  16. Pengembangan Antibodi Poliklonal dari Stadium Oosista, Sporosista, dan Sporozoit Eimeria tenella (THE DEVELOPMENT OF POLYCLONAL ANTOBODY FROM EIMERIA TENELLA OOCYST, SPOROCYST, AND SPOROZOITE STADIUM

    Directory of Open Access Journals (Sweden)

    Galuh Tresnani

    2013-09-01

    Full Text Available The research on developing diagnostic method, vaccine, and drugs for coccidiosis has been focused onthe finding of the immunogenic molecule in Eimeria. The identification of this agent will need the antibodywhich can recognize the biomolecule in the antigen. Antibody that has been developed for this purposeshould be analyzed first, and one of the simple methods for analyzing this antibody is through dot blotanalysis. The objective of this research was to analyze the polyclonal antibody which developed from theoocyst, sporocyst, and sporozoite of  E. tenella using dot blot analysis. The antigen for this polyclonalantibody was made from each of the E. tenella stadium by sonication. Fifteen mice, divided into 3 groups,were then injected subcutaneously with each antigen. The sera from these mice were then collected, analyzedby using ELISA and then it will be used for the dot blot analysis. The research result showed that thepolyclonal antibody which has been developed in mice from each antigen can react with the antigen itself.From this result it can be concluded that the developing of this antibody is successful and it can be used forfurther research in immunoproteomic.

  17. Transfusion management of patients with red blood cell antibodies

    OpenAIRE

    Bujandrić Nevenka B.; Grujić Jasmina N.; Krga-Milanović Mirjana M.

    2013-01-01

    Introduction. Red blood cell antibodies may cause a positive result of pre-transfusion blood compatibility testing (crossmatch test). It can be a problem to provide suitable blood units for patients with clinically significant antibodies to high-frequency antigens as well as for those with multiple alloantibody specificities. This study was aimed at identifying transfused patients in the population of South-Backa who had developed clinically significant red...

  18. Test the Artifact - Develop the Organization

    DEFF Research Database (Denmark)

    Bossen, Claus

    2006-01-01

    Objective and methods: The paper aims to develop further insights into the process of implementationof IT in health care by describing findings from a study of a trial implementationof a newly developed electronic medication plan (EMP) in three hospitals in a county in Denmark.A sociotechnical...

  19. DEVELOPING COMMUNICATIVE LANGUAGE TESTS FOR SENIOR HIGH SCHOOL

    Directory of Open Access Journals (Sweden)

    Y. M. Harsono

    2005-01-01

    Full Text Available The Communicative Approach of teaching English in senior high school in Indonesia has been adopted since the implementation of The 1984 Curriculum, but the tests–the communicative language tests–(CL Tests have not been developed and used properly. The objective of the study is to develop CL Tests for senior high school. The procedure of conducting the study consists of three major steps, that is, identifying the objectives, developing the test specification, and developing the CL Tests. The development of the CL Tests in detail consists of fifteen sub-steps from determining what language skills tested, selecting the suitable source materials, up to rewriting the CL Tests to be used as CL Tests alternative for senior high school. The results of the test development reveal that there are fifteen CL Tests consisting of three tests of listening, three reading, three speaking, and three writing tests. The whole tests have construct and content validity, no complete evidence of concurrent validity with report marks and semester test scores, high to very high inter-rater reliability, and no complete practicality.

  20. Development of Rapid Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Burkholderia pseudomallei

    Science.gov (United States)

    Suttisunhakul, Vichaya; Wuthiekanun, Vanaporn; Brett, Paul J.; Khusmith, Srisin; Day, Nicholas P. J.; Burtnick, Mary N.; Limmathurotsakul, Direk

    2016-01-01

    Burkholderia pseudomallei, the causative agent of melioidosis, is an environmental bacillus found in northeast Thailand. The mortality rate of melioidosis is ∼40%. An indirect hemagglutination assay (IHA) is used as a reference serodiagnostic test; however, it has low specificity in areas where the background seropositivity of healthy people is high. To improve assay specificity and reduce the time for diagnosis, four rapid enzyme-linked immunosorbent assays (ELISAs) were developed using two purified polysaccharide antigens (O-polysaccharide [OPS] and 6-deoxyheptan capsular polysaccharide [CPS]) and two crude antigens (whole-cell [WC] antigen and culture filtrate [CF] antigen) of B. pseudomallei. The ELISAs were evaluated using serum samples from 141 culture-confirmed melioidosis patients from Thailand along with 188 healthy donors from Thailand and 90 healthy donors from the United States as controls. The areas under receiver operator characteristic curves (AUROCC) using Thai controls were high for the OPS-ELISA (0.91), CF-ELISA (0.91), and WC-ELISA (0.90), while those of CPS-ELISA (0.84) and IHA (0.72) were lower. AUROCC values using U.S. controls were comparable to those of the Thai controls for all ELISAs except IHA (0.93). Using a cutoff optical density (OD) of 0.87, the OPS-ELISA had a sensitivity of 71.6% and a specificity of 95.7% for Thai controls; for U.S. controls, specificity was 96.7%. An additional 120 serum samples from tuberculosis, scrub typhus, or leptospirosis patients were evaluated in all ELISAs and resulted in comparable or higher specificities than using Thai healthy donors. Our findings suggest that antigen-specific ELISAs, particularly the OPS-ELISA, may be useful for serodiagnosis of melioidosis in areas where it is endemic and nonendemic. PMID:26912754