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1
Kleiner, Eric J. "Evaluation of Enzyme-Linked Immunosorbent Assay (ELISA) Test Kits for the Quantitative Determination of Endocrine Disrupting Compounds (EDCs) in Aqueous Phase Environmental Samples." University of Cincinnati / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1282056078.
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Mello, Marcelle Bral de. "Padronização do Kit de ELISA Recombinante para o diagnóstico da doença de Chagas visando sua utilização nos Serviços de Hemoterapia." Instituto de Tecnologia em Imunobiológicos, 2009. https://www.arca.fiocruz.br/handle/icict/5875.
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Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.
Bio-Manguinhos produz, desde a década de 80, Kits para diagnóstico, inclusive para o diagnóstico da doença de Chagas. O kit EIE-Recombinante-Chagas, que utiliza na sua composição as proteínas recombinantes CRA e FRAdo T. cruzi, foi registrado como produto junto à ANVISA no ano de 1999. No período de renovação do registro, o Kit passou por uma avaliação prévia no INCQS, onde não foi possível alcançar condição satisfatória quanto aos níveis de sensibilidade. O presente trabalho teve por objetivo retomar a padronização deste Kitno âmbito do desenvolvimento tecnológico. Para tanto foi estabelecido a padronização de três modelos de protótipos, utilizando variações das construções disponíveis das proteínas recombinantes CRA e FRA, na forma de apresentação como antígeno e como conjugado. Posteriormente, foi avaliado o desempenho de cada protótipo, em três diferentes fases. Na primeira fase, o melhor desempenho foi apresentado pelo protótipo 3, cujos níveis de sensibilidade e especificidade foram respectivamente de 99% e 99,5% (IC 95%). Na segunda fase o nível de sensibilidade do protótipo 3 foi de 95.8% e o de especificidade 100%, mantendo a melhor condição de desempenho em relação aos demais. Na última fase, somente o protótipo 3 foi avaliado em função da indisponibilidade de volume das amostras selecionadas e os níveis de sensibilidade e especificidade encontrados de 100%.O desempenho do protótipo 2 foi insatisfatório nas duas primeiras fases de avaliação.Tendo em vista o bom desempenho alcançado pelo protótipo 3, propomos a continuação deste modelo na elaboração futura de lotes-piloto, de estudos multicêntricos e de validação, em ações conjuntas com o controle e a garantia da qualidade para a elaboração de documentação visando pedido de registro desse produto junto aos órgãos regulatórios.
Bio-Manguinhos produces, since the 80’s, a variety of kits for diagnosis, including the kit for Chagas’ disease diagnostic. The EIE-Recombinant-Chagas’ kit, which has in its constitution the recombinant proteins CRA and FRA from T. cruzi, was registered at ANVISA as a product in 1999. During the renewal of the registration period, the kit was submitted to a previous registration analysis by The National Institute for Quality Control in Health (INCQS), when it did not show satisfactory sensitivity levels. The present work aimed to remake the kit standardization process under the scope of the technological development. In order to do it, we established the standardization of three prototype models, each one using variations of available constructions from CRA and FRA recombinant proteins in the form of presentation as antigen and as conjugate. Afterwards, we evaluated the performanceof each prototype, in three different stages. In the first one, prototype 3 showed the best performance with 99% sensitivity and 99.5% specificity levels (CI 95%). In the second stage, prototype 3 levels sensitivity and specificity were 95.8% and 100% respectively, keeping the best performance condition as compared to the other two prototypes. In the last stage, only prototype 3 was assessed due to the lack of sample volumes. The reported sensitivity and specificity levels forsuch prototype were equal to 100%. The performance of prototype 2 was unsatisfactory in the first two assessment stages. Taking into account the performance achieved by prototype 3, we propose the continuation of this model for future development of pilot lots, multicenter and validation studies, in joint actions with quality assurance and quality control for the arrangement of the documents requested by the National Regulatory Agency for the registration of the product.
3
Andersson, Josefin. "Utvärdering av Malaria Antigen ELISA kit för diagnostik av malaria vid Christian Medical College and Hospital i Vellore, Indien. : en jämförande studie mellan Quantitative buffy coat och enzyme-linked immunosorbent assays (ELISA) metodik." Thesis, Örebro universitet, Institutionen för hälsovetenskap och medicin, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:oru:diva-4889.
Повний текст джерелаАнотація:
Malaria är ett globalt hälsoproblem som orsakar många dödsfall runt om i världen varje år och nästan hälften av jordens befolkning ligger i riskzonen att drabbas av sjukdomen. I Indien drabbas mellan 2-3 miljoner människor varje år och det inträffar omkring 900 dödsfall. Malaria orsakas av Plasmodium sp. som är en protozoe, och det finns fyra olika arter som är patogena för människor, P. vivax, P. ovale, P. falciparium samt P. malariae. Vanliga metoder för att diagnostisera malaria är genom tunna och tjocka blodutstryk som färgas till exempel med Giemsa, Fields eller Leishmans färgningsteknik och studeras mikroskopiskt, Quantitative Buffy Coat (QBC), PCR tester, acridinorange färgning samt olika immunologiska tester för detektion av antikroppar eller antigen som till exempel enzyme-linked immunosorbent assays (ELISA) test och dipstick test. Syftet med denna studie är att utvärdera om en användning av SD Bio Line Malaria Antigen ELISA kit ger en mer känslig, tillförlitlig, praktisk samt mindre kostsam diagnostikmetod för malaria hos patienter med misstänkt malariainfektion än den nuvarande guldstandardmetoden, QBC tillsammans med blodutstryk, vid Christian Medical College and Hospital i Vellore. Patientproverna har i både ELISA testet samt QBC testet tillsammans med utstryk erhållit samma resultat vilket tyder på att SD Bio Line Malaria Antigen ELISA kitet skulle kunna vara en lika bra diagnostikmetod som QBC testet för diagnos av malaria. ELISA kitet har dock fler nackdelar, i jämförelse med QBC testet, så därför är slutsatsen att SD Bio Line Malaria Antigen ELISA kitet inte är en mer lämplig diagnostisk metod för malaria än den som används vid CMCH. Men då ELISA testet ändå ger en säker diagnos, enligt resultatet i studien, kan den vara ett lämpligt test inom något annat användningsområde.
4
Wesolowski, Eva. "Monoclonal antibody enzyme-linked immunosorbent assay for carcinoembryonic antigen." Thesis, McGill University, 1987. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=66230.
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5
Kiote-Schmidt, Chrissoula. "Etablierung eines kompetitiven enzymgekoppelten Immunoassays zum Nachweis eines kleinen Peptids in Serum- und Liquorproben." [S.l. : s.n.], 2007. http://nbn-resolving.de/urn:nbn:de:bsz:289-vts-59192.
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6
Kim, In Soo. "A quantitative enzyme linked immunosorbent assay for polychlorinated biphenyls in transformer oil." Thesis, Cranfield University, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.323838.
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7
Meindel, Mandy J. "Enzyme-linked immunosorbent assay to measure serum ferritin in toucans (Ramphastidae sp.)." Thesis, Kansas State University, 2015. http://hdl.handle.net/2097/18896.
Повний текст джерелаАнотація:
Master of ScienceDepartment of Diagnostic Medicine/ Pathobiology
Lisa M. Pohlman
Background: Iron storage disease has proven to be a serious health concern for captive toucans. Physiologic mechanisms to efficiently extract iron from naturally iron-deficient diets appear the likely cause of iron overload when fed iron-sufficient diets in captivity. Iron overload can result in diabetes, heart failure, and even death. Serum ferritin concentrations are considered the most reliable screening tool to predict total body iron stores in many species, but an assay has not been available to measure serum ferritin in toucans. Objective: The purpose of this study was to develop an enzyme-linked immunosorbent assay (ELISA) to measure serum ferritin in toucans using a polyclonal antibody in a sandwich arrangement. Methods: Ferritin was isolated from toucan liver and used as a standard. A rabbit polyclonal anti-toucan antibody was used as the capture antibody and as a detection antibody conjugated to horseradish peroxidase. Linearity of toucan ferritin standards, effect of serum dilution, recovery of added ferritin standards, and intra- and inter-assay variability were determined. Results: Ferritin standards were linear from 0 to 50 ng/ml. The relationship between serum dilution and serum ferritin concentration was also linear. When 10, 20, 30, 40, or 50 ng/ml of purified toucan ferritin were added to diluted serum, the recoveries varied from 69% to 104%. The intra-assay variability for four test serum samples averaged 11% and the inter-assay variability for the same four samples averaged 11%. Conclusions: Although the results from the linearity and recovery studies are promising for assay development when viewed independently, preliminary ferritin concentrations from all toucans studied are much higher than expected. Upon further evaluation including Dot blot assays, Western blot assays, SDS-PAGE, and protein determination of the ferritin stock solution, it was determined that the ferritin stock solution did not contain a pure protein and therefore likely renders the assay invalid. Further testing is needed to confirm these findings.
8
Leung, Sau-man Sally. "Serodiagnostic utility of an ELISA assay based on a recombinant antigen MP1 of penicillium marneffei." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B2337309X.
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9
Carroll, Andrea D. "Development of bead injection methodology for immunoassays /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/8599.
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10
Kwok, Siu Kwong Alan. "Enzyme-linked immunosorbent assay development for advanced glycation end products and brochocin-C." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp05/mq22621.pdf.
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11
Campbell, Charles Nelson. "Separationless immunoassay and DNA sensing using wired enzyme based amperometric affinity electrodes /." Digital version:, 2000. http://wwwlib.umi.com/cr/utexas/fullcit?p9992761.
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12
Murphy, Jamie Ellen. "Development and evaluation of a sarcocystis neurona-specific IgM capture enzyme linked immunosorbent assay." The Ohio State University, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=osu1399903151.
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13
Nitaya, Poopyruchpong Vithoon Viyanant. "Detection of opisthorchis viverrini infection by enzyme-linked immunosorbent assay using partially purified antigens /." abstract, 1988. http://mulinet3.li.mahidol.ac.th/thesis/2531/31E-Nitaya-P.pdf.
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14
Östling, Jeanette. "An Approach to Improve the Detection System of a Diagnostic Enzyme-Linked Immunosorbent Assay." Thesis, KTH, Skolan för bioteknologi (BIO), 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-214620.
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15
Orchard, Robert Graham. "A flow-through enzyme-linked immunoassay for progesterone." The University of Waikato, 2007. http://hdl.handle.net/10289/2277.
Повний текст джерелаАнотація:
Bovine reproductive performance is one of the most important factors influencing dairy farm profitability. Present-day techniques for oestrus- and pregnancy-detection are unreliable and labour-intensive. Although measuring milk-progesterone at regular intervals allows the fertility status of a cow to be determined reliably, the labour cost of collecting and analysing samples is prohibitive. This project aimed to develop a progesterone sensing system that could be automated and integrated with the milking unit, thus minimising labour costs. The proposed system involved mixing the milk sample with an enzyme-antibody conjugate and then passing the sample through a column containing immobilised progesterone. Any progesterone in the milk would inhibit conjugate binding to the column. An enzyme substrate would then flow through the column and bound conjugate would be detected as a colour change at the column's outlet. Periodate-coupling was used to attach horseradish peroxidase enzyme to anti-progesterone antibody, and progesterone-3-carboxymethyloxime was immobilised on the polystyrene bead surface using amine-coupling. Both techniques are widely used. Initial experiments attempted to verify the success of these two reactions simultaneously, whereas later experiments focused on the bead-coating. Beads were suspended in a specially-constructed syringe and the antibody activity of the eluted solution was measured by SPR. However, a combination of non-specific binding and antibody stability and activity issues meant neither reaction was conclusively verified. Many trials were done to investigate how to overcome the problems encountered but a suitable, workable procedure was not developed. Despite poor progress, the problems encountered did not undermine the project's potential. There remains optimism of developing an on-line method if research were to continue.
16
Lövgren, Ulf. "Enzyme immunoassay in combination with liquid chromatography for sensitive and selective determination of drugs in biosamples." Lund : Dept. of Analytical Chemistry, Lund University, 1997. http://books.google.com/books?id=Ju1qAAAAMAAJ.
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17
梁秀雯 and Sau-man Sally Leung. "Serodiagnostic utility of an ELISA assay based on a recombinant antigen MP1 of penicillium marneffei." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31970096.
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18
de, Alwis Wathuthanthirige Uditha. "Analytical applications of chemically modified antibodies." Diss., The University of Arizona, 1988. http://hdl.handle.net/10150/184601.
Повний текст джерелаАнотація:
The components involved in an immunoassay were investigated in order to improve the detection limits of the ELISA and to make the assay adaptable to a flow injection analysis (FIA) configuration. The goal being the total automation of the ELISA procedure which is long, tedious and has high standard deviation. The antibody purification and cleavage methods were studied with special emphasis on obtaining products with highest immunological activity. The antibody-enzyme coupling reactions using homobifunctional reagents and heterobifunctional reagents were studied in order to attempt the preparation of highly characterized reagents. The fragments of IgG were coupled to polymeric supports via the hinge thiol groups to retain the maximum immunological activity. This method was found to be superior to those methods involving coupling via amino group. These reagents were used in the development of a sandwich ELISA for bovine IgG. The range of assay was in the 20-1000 femtomole range with a linear dynamic range of 2 orders of magnitude and an accuracy of 2-5%. A competitive ELISA based on the use of immobilized anti-human IgG Fab' fragments was developed. The linear dynamic range for this assay was found to be less than one order of magnitude. The detection limit was in the low picomole range with an accuracy of 2-5%. Based on the principle used in the two assays an enzyme immobilization scheme was developed for the reversible immobilization of these enzymes. Which was subsequently utilized in the determination of substrate in the picomole range in a reagent less FIA technique. The goals of this research project were realized in that the FIA system utilized in this work was capable of carrying out totally automated ELISA assays with an accuracy far surpassing the conventional plate ELISA assays.
19
Sheehan, C. P. "The application of the enzyme-linked immunosorbent assay (ELISA) to ABH grouping in forensic science." Thesis, University of Surrey, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.381661.
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20
Cheow, Lih Feng. "Development of an Enzyme-Linked ImmunoSorbent Assay (ELISA) with Enhanced Sensitivity in a Nanofluidic System." Thesis, Massachusetts Institute of Technology, 2009. http://hdl.handle.net/1721.1/55148.
Повний текст джерелаАнотація:
Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2009.Cataloged from PDF version of thesis.
Includes bibliographical references (p. 66-68).
Experimental studies were performed to evaluate the kinetics and equilibrium binding constants of biomolecules in nanofluidic channels. Binding events in the nanochannel were detected using electrical and fluorescence methods. We concluded that antibody-antigen binding constants in nanochannels were similar to experiments performed in microtiter plates at low antigen concentrations; however the bound fraction in nanochannels at high antigen concentration decreased due to steric hindrance. Binding kinetics in nanochannels was limited by convective transport of analytes, instead of diffusion or reaction. We also found that enzymatic reactions in nanochannels were very effective due to short diffusion length and high surface area to volume ratio. A bead based ELISA was developed to exploit the rapid binding reactions in the bulk and efficient enzymatic conversion in the nanochannels. Additionally, electrokinetic concentrators were integrated with multiplexed bead based ELISA to further improve the detection sensitivity of a sandwich immunoassay.
by Lih Feng Cheow.
S.M.
21
Lupica, Samuel J. "Nitrate Toxicity to Common Carp Measured Noninvasively by Novel Enzyme-linked Immunosorbent Assay for Cortisol." University of Toledo Health Science Campus / OhioLINK, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=mco1226956326.
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22
Botha, Elizabeth Magdelena. "Molecular characterization of South African lineage II West Nile virus isolates ltime PCR assay." Pretoria : [s.n.], 2008. http://upetd.up.ac.za/thesis/available/etd-06122008-130924/.
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23
Gossen, Ulrike. "Indikation der Bodenqualität mittels immunologischer Quantifizierung der Enzymkonzentration am Beispiel der Urease in schwermetallkontaminierten Böden /." Großbeeren [u.a.] : IGZ, 2002. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=010272469&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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24
Lack, Pamela. "Bestimmung von Immunglobulin G und M im Serum neugeborener Kälber während der ersten zehn Lebenstage unter besonderer Berücksichtigung des Fütterungsregimes." Giessen : VVB Laufersweiler, 2006. http://geb.uni-giessen.de/geb/volltexte/2007/4107/index.html.
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25
Liu, Huiqing. "Characterization of two-component organothiol mixed monolayers on gold and quantification of nonspecific adsorption on mixed SAM biosensor platforms using electrochemical enzyme immunology." Auburn, Ala., 2007. http://repo.lib.auburn.edu/2007%20Spring%20Theses/LIU_HUIQING_29.pdf.
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26
Tse, Maying Tsemay, and 謝美盈. "Evaluation of three commercially available influenza A type-specific blocking enzyme-linked immunosorbent assays for seroepidemiologicalstudies of influenza A virus infection in pigs." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B4827382X.
Повний текст джерелаАнотація:
The emergence of the pandemic H1N1 2009 virus of swine-origin and its transmission back to swine highlighted the need for global surveillance of swine influenza. Serology can help to address the epidemiological situation of influenza infection. Since typical serology tests such as hemagglutination inhibition or microneutralization assays are subtype and partially virus-lineage specific, it is important to select appropriate viral antigens for such studies. A poorly chosen panel of antigens will lead to underestimation of the seroprevalence. The choice of well-matched antigen is difficult if there is no prior virological surveillance in that area and even if there was virological surveillance data, transient infections may go undetected. Hence an universal influenza A type reactive serological test is needed.
While such tests are available for poultry, there is little published data on the performance of these commercial influenza ELISA assays for serology on swine sera. In this study we evaluated 3 commercially available competitive ELISA assays, IDEXX? Influenza A Ab test, IDEXX? AI MultiS-Screen Ab Test and IDVet ID Screen? Influenza A Antibody Competition ELISA kit for detecting influenza type A reactive antibodies in swine. The virus antigens and the serum samples were obtained from a 14-year systematic abattoir-based virological and serological surveillance for swine influenza in southern China. The performance was evaluated by ROC curve and scatter plot, together with other statistical parameters including the Youden index to optimize the cut-off levels. Using the optimized cut-off levels, sensitivity and specificity of the IDEXX? Influenza A Ab test was 86% and 89% respectively; for IDEXX? AI MultiS-Screen Ab Test was 91% and 87% and for IDVet ID Screen? Influenza A was 95% and 79%, respectively. These findings help to provide different cut-off levels to maximize the sensitivity or specificity to suit different purposes. We found that the ELISA assay was useful in detecting serum samples that may be positive for influenza antibody but missed in the serology screening tests due to limitations in the chosen antigen panel. The ELISA assay maybe helpful in global swine influenza surveillance programs.published_or_final_version
Microbiology
Master
Master of Medical Sciences
27
Eggemann, Gabriele. "Untersuchungen zum Vorkommen von Chlamydieninfektionen in Zuchtsauenbeständen und deren Bedeutung für das Fruchtbarkeitsgeschehen /." Giessen : Köhler, 1999. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=008937022&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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Jahn, Carsten. "Analytik cuticulagebundener Rückstände des Fungicids Chlorthalonil unter besonderer Berücksichtigung immunchemischer Nachweisverfahren /." Aachen : Shaker, 2000. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=009061215&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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29
Hitt, John Michael 1952. "DETERMINATION OF THE EFFICACY OF THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) IN CHARACTERIZING CROTALUS SNAKE VENOM AT THE SPECIES LEVEL." Thesis, The University of Arizona, 1986. http://hdl.handle.net/10150/276352.
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30
Leelayawat, Chanvit. "Evaluation of antibody elution techniques using enzyme-linked antiglobulintest (ELAT) /." abstract, 1986. http://mulinet3.li.mahidol.ac.th/thesis/2529/29E-Chanvit-L.pdf.
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31
Goodall, Stephen Francis. "The preparation and characterisation of an enzyme-monoclonal antibody conjugate : a thesis." Thesis, Queensland University of Technology, 1989. https://eprints.qut.edu.au/35948/1/35948_Klick_1989.pdf.
Повний текст джерелаАнотація:
Potentially life threatening thromboembolic disorders can be detected and monitored by determining the level of D-Dimer in human plasma and serum. Assays employing antibodies provide sensitive and highly specific analytical methods for biological analytes. The use of monoclonal antibody technology provides the basis for the D-Dimer Enzyme ImmunoAssay manufactured by AGEN Biomedical Ltd. The assay uses an enzyme-antibody conjugate for the detection of an analyte immunologically
bound to the well of a microtitre plate. This conjugate consists of a specific monoclonal antibody bound to the enzyme, horseradish peroxidase(HRPO). Evaluation of the periodate conjugation procedure used in the commercial preparation of the assay indicated that this method
was unsatisfactory for routine production of HRPO-antibody conjugates because of inconsistent
yields and variable activities of the conjugate. The heterobifunctional reagent N-succinimidyl
3-(2- pyridyldithio)propionate (SPDP) was evaluated as a cross-linking reagent for HRPO-antibody
conjugation. The conjugation method was developed with a series of characterization tests to monitor the reactions used in the conjugation and the performance of the final product. A conjugation method was developed which resulted in a conjugate with an activity approximately five times that of the periodate conjugate, a variation in the conjugate activity of only 22% between
preparations conjugates and equivalent D-dimer analyses for human plasmas. The SPDP conjugation method provided a routine procedure for the consistent preparation of a commercial conjugate consisting of HRPO and an anti D-dimer monoclonal antibody.
32
Leitch, Robert A. "Analysis of immunodominant epitopes of Neisseria gonorrhoeae lipopolysaccharide using a competitive inhibition enzyme linked immunosorbent assay." Thesis, University of Ottawa (Canada), 1985. http://hdl.handle.net/10393/4930.
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33
Booth, Ronald A. Carleton University Dissertation Biology. "Development of cloth-enzyme immunoassays for the detection of E. coli O157 and verotoxin." Ottawa, 1996.
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34
Johnson, Raymond Camille Joseph. "Detection of nepoviruses by ELISA in tissue-cultured and field-grown grapevines." Thesis, University of British Columbia, 1988. http://hdl.handle.net/2429/27968.
Повний текст джерелаАнотація:
The detection by serology of nematode-transmitted polyhedral viruses (nepoviruses) in grapevines is often unreliable. Nepoviruses were detected by enzyme-linked immunosorbent assay (ELISA) in tissue-cultured and field-grown grapevines. Nepovirus detection in in vitro plants (plantlets) was affected by virus distribution and growth room temperature. The reliability of virus detection in field-grown grapevines was improved when modified grinding buffers were used.
Arabis mosaic virus (AMV) was detected by ELISA, for the first time, in in vitro grapevines initiated from field-and screenhouse-grown plants throughout the summer. The virus was not reliably and repeatedly detected in in vitro plantlets grown at 25°C.
AMV and grapevine fanleaf virus (GFLV) distribution was not uniform throughout the plantlets. This distribution was affected by the culture room temperature. The best plant parts to sample for virus detection came from the zones of rapidly proliferating shoots. The viruses were sometimes not detected in samples taken from other tissues.
Growth room temperature had an important effect on virus detection in plantlets. The highest virus titres were found in plants growing at 15°C. Temperature increases in 5°C steps to 30°C reduced virus titre. AMV became undetectable in nearly all plantlets growing at 30°C for as little as 30 days. Growth at 30°C reduced ELISA absorbance values by 76% after 8 days and after 21 days the values were at 4% of pre-treatment levels. The virus titre dropped below detectable levels in most plantlets. AMV could not be detected in plantlets or rooted explants after being placed in a 30°C treatment for 2 months.
Tomato ringspot virus was detected by ELISA, for the first time, in in vitro grapevine plants. The virus was repeatedly detected in in vitro plants growing at 20°C.
Under the typical summer conditions experienced at Sidney, B.C., modifying the standard ELISA grinding buffer (0.01 M phosphate buffered saline, pH 7.4, 0.05% Tween-20, 0.2% ovalbumin, 2% polyvinylpyrrolidone) was essential for reliable detection of AMV or GFLV. AMV was reliably detected by ELISA in foliar samples from field or screenhouse plants throughout the summer when the grinding buffer was modified by increasing the pH to at least 8.2 and adding 1% nicotine or 0.15 M phosphate buffered saline. The most reliable results with GFLV were obtained with the nicotine enhanced buffers.
In comparison, because of the increased workload associated with growing plants in vitro and the unreliable detection of viruses in these plants, it remained preferable to detect nepoviruses in field plants by ELISA.Land and Food Systems, Faculty of
Graduate
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Sattler, Tatjana, Eveline Wodak, Sandra Revilla-Fernández, and Friedrich Schmoll. "Comparison of different commercial ELISAs for detection of antibodies against porcine respiratory and reproductive syndrome virus in serum." Universitätsbibliothek Leipzig, 2015. http://nbn-resolving.de/urn:nbn:de:bsz:15-qucosa-158536.
Повний текст джерелаАнотація:
Background: In recent years, several new ELISAs for the detection of antibodies against the porcine reproductive and respiratory disease virus (PRRSV) in pig serum have been developed. To interpret the results, specificity and sensitivity data as well as agreement to a reference ELISA must be available. In this study, three commercial ELISAs (INgezim PRRS 2.0 - ELISA II, Priocheck® PRRSV Ab porcine – ELISA III and CIVTEST suis PRRS E/S PLUS - ELISA IV, detecting PRRSV type 1 antibodies) were compared to a standard ELISA (IDEXX PRRS X3 Ab Test - ELISA I). The serum of three pigs vaccinated with an attenuated PRRSV live vaccine (genotype 2) was tested prior to and several times after the vaccination. Furthermore, serum samples of 245 pigs of PRRSV positive herds, 309 pigs of monitored PRRSV negative herds, 256 fatteners of assumed PRRSV negative herds with unknown herd history and 92 wild boars were tested with all four ELISAs. Results: ELISAs II and III were able to detect seroconversion of vaccinated pigs with a similar reliability. According to kappa coefficient, the results showed an almost perfect agreement between ELISA I as reference and ELISA II and III (kappa > 0.8), and substantial agreement
between ELISA I and ELISA IV (kappa = 0.71). Sensitivity of ELISA II, III and IV was 96.0%, 100% and 91.5%, respectively. The specificity of the ELISAs determined in samples of monitored PRRSV negative herds was 99.0%, 95.1% and 96.4%, respectively. In assumed negative farms that were not continually monitored, more positive samples were found with ELISA II to IV. The reference ELISA I had a specificity of 100% in this study. Conclusions: All tested ELISAs were able to detect a PRRSV positive herd. The specificity and sensitivity of the tested commercial ELISAs, however, differed. ELISA II had the highest specificity an ELISA III had the highest sensitivity in comparison to the reference ELISA. ELISA IV had a lower sensitivity and specificity than the other ELISAs.
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Bates, Kimberly M. "The biology of dictyocaulus viviparus in Missouri /." free to MU campus, to others for purchase, 1997. http://wwwlib.umi.com/cr/mo/fullcit?p9841262.
Повний текст джерела
37
Dell, Kerstin. "Entwicklung eines indirekten ELISA zum serologischen Nachweis von Opisthorchis felineus (Rivolta, 1884) und Metorchis bilis (Braun, 1790) (Trematoda: Opisthorchiidae) beim Fuchs : mit einem Beitrag zu den Leberenzymaktivitäten bei experimenteller Opisthorchiidose /." Friedland : Bielefeld, 2001. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=009538526&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
Повний текст джерела
38
Tandy, Mark Wayne. "Early diagnosis of systemic Candida infection : development of an enzyme immunoassay for mycelial antigen and comparison with other methods." Thesis, Queensland University of Technology, 1986. https://eprints.qut.edu.au/36810/1/36810_Tandy_1986.pdf.
Повний текст джерелаАнотація:
An Enzyme immunoassay (ELISA) using monoclonal antibodies was developed to detect antigen associated with Candida albicans mycelial cytoplasm. One hundred and fourteen sera from patients with and at risk of invasive Candida infection were assayed by this and five other laboratory diagnostic techniques; Counterimmunoelectrophoresis, Latex agglutination for mannan antigen, Passive haemagglutination for mannan antibody, Serum arabinitol levels, and Serum arabinitol/creatinine ratios. Sensitivites of the five tests were 22%,11%,44%,89%,and 78% respectively. Sensitivity for the ELISA was 44%. Specificities for the five tests ranged from 84% to 100%. Specificity for the ELISA was 100%, being positive for 4 of 9 systemically infected patients and none of 12 colonised and 83
non infected patients (chi-square=43.9; df=l; P=0.0000). This suggests that detection of antigens of mycelial cytoplasm origin are related to invasive disease. The low sens ivity of the ELISA test may be improved by better definition and characterisation of the antigens associated with invasive Candida infection and production of specific antibodies directed against them. Comparison of all the above tests for the detection of patients suffering systemic Candida infection showed that screening of patients using serum arabinitol levels and confirmation of positive results with serum arabinitol/creatinine ratios offered the best approach for laboratory diagnosis of invasive disease.
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Kellerer, Thomas Georg. "Entwicklung von immunochemischen und PCR-Methoden zum qualitativen Nachweis von Tilletia-Arten und Ustilago nuda in Saatgut." kostenfrei, 2009. https://mediatum2.ub.tum.de/node?id=739743.
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Sylvestre, Tatiane Fernanda [UNESP]. "Avaliação da acurácia do teste imunoenzimático e de sua contribuição no seguimento de pacientes com paracoccidioidomicose em tratamento e no diagnóstico de doença reativada." Universidade Estadual Paulista (UNESP), 2013. http://hdl.handle.net/11449/108556.
Повний текст джерелаАнотація:
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000753373.pdf: 543622 bytes, checksum: 568807a679f23679d4c54335db59a4d0 (MD5)O reaparecimento de manifestações clínicas após tratamento eficaz, neste texto identificado como recaída, tem sido pouco avaliado. Assim, foram estudados os casos de recaída observados em 400 pacientes, 93 com a forma aguda/subaguda (FA) e 307 com a crônica (FC), que já tinham apresentado cura aparente, isto é, com cura clínica, normalização da velocidade de hemossedimentação e cura sorológica – caracterizada pela presença de teste negativo à imunodifusão dupla em gel de agar por dois anos. Vinte e um (5,2%) desses pacientes apresentaram recaídas da doença. Três (14,3%) eram do sexo feminino e 18 (85,7%) do masculino, com razão de masculinidade de 6:1. Dos 21 pacientes com recaída, 15 (4,8%) apresentavam a FC e 6 (6,4%) a FA (p>0,05). As reativações ocorreram de 46 a 296 meses após introdução do tratamento (Md=96) e de 4 a 267 meses (Md= 60) após sua suspensão. As formas clínicas não diferiram quanto aos tempos decorridos até a reativação. O diagnóstico sorológico de recaída pela IDD foi feito em apenas 45% dos casos, o que levou à avaliação de outros testes para esse fim. Assim, foi realizado o enzimaimunoensaio (ELISA) e o immunoblotting (IB). A sensibilidade da IDD e do ELISA / 0,710 foi 76,1% em amostras de soro obtidos no pré-tratamento (p=0,25). No diagnóstico de recaída, a sensibilidade da IDD foi menor que no pré-tratamento (80% vs 45%; p=0,017), enquanto o ELISA / 0,710 foi igual (80% vs 65%;p=0,125). A sensibilidade do IB para diagnóstico de recaída foi de 12,5% na identificação da gp70 e 43,8% na da gp43 (p<0,05). A avaliação da acurácia do teste ELISA revelou sensibilidade de 96%, especificidade de 95%, valor preditivo positivo de 95%, valor preditivo negativo de 96% e acurácia de 95,5% quando o cut-off utilizado foi a densidade óptica de 0,710, obtido em função da construção da receiver operator characteristc – ROC, para um intervalo de confiança ...
The reappearance of clinical manifestations after efficacious treatment, here identified as relapse, has been rarely evaluated. Thus, the cases of relapse observed in a cohort of 400 patients, 93 with acute/subacute form (AF) and 307 with chronic form (CF) were studied. They had already reached apparent cure, characterized as clinical cure, normalization of the erythrocyte sedimentation rate and serological cure, with a negative double agar gel immunodiffusion test (DID) for two years after antifungal discontinuation. Twenty-one (5.2%) of these patients had relapses. Three (14.3%) were female and 18 (85.7%) were male, with a male:female ratio of 6:1. Of the 21 relapsed patients, 15 (4.8%) presented the CF and 6 (6.4%) the AF (p>0.05). The relapse occurred 46-296 months after introduction of the treatment (Md=8 yrs) and from 4 to 267 months (Md=5 yrs) after withdrawal. Clinical forms did not differ regarding to the time elapsed until relapse. DID was positive in only 45% of the relapsed cases, which led to the evaluation of other tests to diagnose this condition. Thus, the enzyme-linked immunosorbent assay (ELISA) was standardized and the cut off was determined using the curve receiver operator characteristic – ROC and confidence intervals of 95% and 99%, showing optical densities of 0.710 and 0.850, respectively. Then, serological evaluation was performed using ELISA/0.710 and ELISA/0.850, and immunoblotting identifying gp43 (IBgp43) and gp70 (IBgp70). ELISA 0.710 and DID showed the same sensitivity, 76.1%, in serum samples obtained before treatment (p=0.25). DID sensitivity was lower at relapse than before the initial treatment (45% vs 80%; p=0.017), whereas ELISA/0,710 was the same (65% vs 80%; p=0.125). IBgp70 showed a 12.5% and IBgp43 a 43.8% sensitivity for diagnosing relapse (p<0.05). ELISA/0.710 showed a 96% sensitivity, 95% specificity, 95% positive predictive value, 96% negative ...
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Lin, Yuh-Fen, та 林毓芬. "Development of Enzyme-Linked Immunosorbent Assay Residual Detection Kit for β-lactams". Thesis, 2007. http://ndltd.ncl.edu.tw/handle/60126167905599965859.
Повний текст джерелаАнотація:
碩士中興大學
獸醫學系暨研究所
95
The aim of this study is to develop a simple, rapid, and reliable enzyme-linked immunosorbent assay (ELISA) method for detecting the residues of β-lactams and as amoxicillin (AMX), ampicillin (AM), and penicillin G (PG) in the varietal samples. Using the carbodiimide coupling (CD), glutaraldehyde coupling (GA), and physiological coupling (PH) methods, the AMX, AM, and PG was productively conjugated with human serum albumin (HSA) and bovine serum albumin(BSA), respectively. According to the different absorption characteristics of β-lactams, BSA, HSA, and β-lactam-conjugated proteins, the three drug-carrier protein conjugations made using three different conjugated techniques were able to be identified by the high performance liquid chromatography (HPLC) with ultraviolet detector. All conjugated proteins were individually used for producing polyclonal antibodies in rabbits. The results showed that the titers of the serum antibodies from rabbit were 65,536-fold after five boosts. According our results, we suggested that the PH method was the most suitable technique to form the conjugated protein for producing polyclonal antibody. At the same time, we found that BSA was better than HSA as the carrier protein for β-lactams. We boosted the β-lactam-conjugated proteins by using the PH methods to producing the monoclonal antibody. The results showed that the titers of the serum antibodies from mice were 32,768-fold after seven boosts. From the mAb tests, our results indicated that the positive hybridoma ratio for anti-AMX, anti-AM, and anti-PG was 1.96%, 1.88%, and 1.67%, respectively. The lowest detection limit (LDL) of our developed ELISA kit for the AMX, AM, and PG in PBS, milk, fish, chicken, beef, and pork meats were (50, 50, 50, 50, 100 and 100) ng/mL, (0.5, 0.5, 0.5, 0.5, 10 and 10) µg/mL, and (50, 100, 50, 50, 100 and 100) ng/mL, respectively. The coefficient variation value of the intra-assay and inter-assay of our AMX, AM, and PG-ELISA kits were (3.72% and 6.4%), (8.83% and 11.29%), (10.27% and 9.0%), respectively. We found the cross-reactive ratio of the AMX, AM, and PG-ELISA plates for tested quinolones were below 0.01%. From above assay results, our developed ELISA kits seemed to be a simple, rapid, and reproducible method to detect the residues of the tested β-lactams in the chicken and fish meats.
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Liu, An-Chi, and 劉銨錤. "Development of Enzyme-Linked Immunosorbent Assay ( ELISA ) Kit for Detection of BEF Antibodies." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/23530324979948094698.
Повний текст джерелаАнотація:
碩士國立屏東科技大學
獸醫學系所
96
Bovine ephemeral fever virus (BEFV) is classified under the Ephemerovirus genus of the Rhabdoviridae family. It causes an acute febrile disease in cattle and water buffalo. The G glycoprotein is one of five structural proteins of BEFV and is the major protective antigen. The clinical symptoms of this disease are the sudden onset of fever, stiffness, lameness, serous oral and nasal discharges, joint pain, depression, and reduced milk production. It is of major economic importance to the dairy industry. The major means of preventing BEF is through vaccination and monitoring the antibody titer to make sure that the cattle have optimal immune status to resist the disease. The traditional method for BEFV detection is serum neutralization test which is both time-consuming and cumbersome when a lot of samples have to be analysed. In this study, the G1 epitope of the glycoprotein G was expressed by E. coli expression system. The G1 fusion protein was analyzed by SDS-PAGE and its antigenicity demonstrated by Western blot. The purified proteins were used as antigens for development an ELISA kit. BALB/c mice were immunized with BEFV, and the conformation and antigenicity of the G1 fusion protein was proved by Western blot. positive/negative ratio (P/N) of box titration is significance level. Our results demonstrated that the G1 fusion protein can be a good candidate for development of a diagnostic kit. Finally, the G1 fusion protein was used to detect BEF serum from clinical samples in Taiwan. The results also showed that the newly developed ELISA kit have high sensitivity.
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Yiing, Shyh-Shuan, and 殷世栓. "Development of enzyme-linked immunosorbent assay diagnostic kit for fowl cholera in Taiwan." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/14044381933318672448.
Повний текст джерелаАнотація:
碩士國立屏東科技大學
獸醫學系
93
The purpose of this research is to develop an ELISA kit applied to dia- gnose the fowl cholera. CHC-00*01 antigen which is the combination of CHC-00 and CHC-01 strain through Indirect ELISA Test, is compared with the antigen mixed with KSC-98 or KSD-99 strain, in order to detect the ab- sorbent rate of the fowl serum. The result of the biochemical evaluation uti- lizing the Biolog''s GN2 MicroPlateTM points out that 99% are Pasteurella multocida bacterium. The cross infective test shows the interactive infection of P. multocida strain between the chicken and the duck. Box titration is uti- lized to gain the ratio of the antigen and the antibody, and is applied 96-wells ELISA plate as the positive and negative controlling group. The final result indicates that, based on 625 nm wave length, P. multocida binary antigen the absorbent rate is 0.25, is diluted into 1：512 and it is the best ratio that the serum from the chicken is diluted into 1：64. According to the ratio, the OD ( optical density ) value of the positive controlling group of the chicken seru- m is 0.416; the negative one is 0.100. The ratio of the positive and negative of the chicken serum is 0.416/0.100＝4.16≥ 4, which is the distinctive varie- ty. Duncan''s multiple range test for variable is applied for analyzing the resu- lt and it shows that there are the distinctive variety ( P＜0.01 ) between CHC -00*01 antigen and CHC-00*01*KSC-98 antigen; the distinctive variety bet- ween ( P＜0.01 ) CHC-00*01 antigen and CHC-00*01*KSD-99 antigen.The result of applying the ELISA kit to detect the serum of 200 chickens from the fields provides that the positive rate of the chicken serum is 29.5% ( 59/200 birds ) and it is coordinated with the clinical symptom. After 141 known gro- ups with the negative serum sample are tested, if the range of S/N ratio is on 1.0; the reliability of the positive and negative serum is 99% confidence level. i. e., the single absorbent rate is diluted into 1：64; S/N ratios is limited on ＞1.0; defined as a positive serologic response; with the acknowledgment of a 1% false positive error rate at S/N＝1.0. This homologous antigen which is the combination of CHC-00 and CHC-01 strain could be the best antigen for preparing ELISA Kit in improving the diagnosis of fowl cholera immunorea- ctions or infections of chickens in Taiwan.
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Chen, Bo-Rui, and 陳柏叡. "Development of Enzyme-Linked Immunosorbent Assay Residual Detection Kit for Ciprofloxacin and oxolinic acid." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/12793612958520710832.
Повний текст джерелаАнотація:
碩士中興大學
獸醫學系暨研究所
95
The aim of this study is to develop a simple, rapid, and reliable enzyme-linked immunosorbent assay (ELISA) method for detecting the residues of ciprofloxacin (Cipro)and oxolinic acid (OA)in the varietal samples. By using the N-hydroxysuccinimide ester method, the Cipro and OA was productively conjugated with HSA, BSA, and HRP, respectively. Successfully, the tested conjugations were able to be detected by the HPLC with UV and fluoresce detectors. The study showed that the titers of the serum antibodies from rabbit and mice were 65,536-fold and 16,384-fold after six boosters, respectively. The lowest detection limit (LDL) of the Cipro and OA(polyclone and monoclone antibody) in PBS was 0.78 ng/mL and 0.56 ng/mL by the ELISA method, respectively. The coefficient variation value of the intra-assay and inter-assay of the polyclonal and monoclonal antibodies (mAb) Cipro ELISA kit were (5.82%, 7.38%) and (9.58%, 7.46%), respectively. According our results, the LDL of the Cipro ELISA and OA ELISA kits in milk, Fetal Bovine Serium, fish, pork, chicken, shrimp, and beef meats were (1.08 ppb, 0.4 ppb) , (1.37 ppb and 0.47 ppb), (0.83 ppb and 0.9 ppb), (1.1 ppb and 0.5 ppb), (0.88 ppb and 0.43 ppb), (1.32 ppb and 1.17 ppb), and (1.14 ppb and 0.54 ppb), respectively. Meanwhile, our data shown that the cross reaction ratioes of the both kits to frequently used chemical agents were below 1%, except the Cipro ELISA Kits to enrofloxacin (58%) and norfloxacin (37%). From the mAb tests, our results indicated that the positive hybridoma ratio for anti-Cipro and anti-OA was 4.16% and 3.01%, respectively. The coefficient variation values of the intra-assay and inter-assay of OA ELISA kit and mAb OA ELISA kit were (8.19%, 6.706%) and (8.20%, 11.76%), respectively. Our result indicate the LDL of the mAb-Cipro ELISA and mAb-OA ELISA kits in milk, Fetal Bovine Serium, fish, and pork were (0.93 ppb, 0.37 ppb) , (1.03 ppb and 0.43 ppb), (0.87 ppb and 0.86 ppb), (0.85 ppb and 0.51 ppb), respectively. We found the cross-reactive ratio of the OA ELISA plate to our tested non-quinolones was below 0.01%. Our results suggested the LDL of the mAb Cipro and mAb OA in PBS were 0.82 ppb and 0.4 ppb. Meanwhile, the cross-reacted ratio of the bothe mAb Cipro and mAb OA ELISA plates to our tested agents was 0.005. Therefore, our developed Cipro and OA ELISA kit had very good precision. According to these results, our developed Cipro ELISA kit seemed to be a simple and reproducible tool for Cipro residue detection for the different animal productions.
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Shih, Ching-Wen, and 施清文. "Development and Application of the Enzyme-Linked Immunosorbent Assay Residual Detection Kit for Fluoroquinolones." Thesis, 2003. http://ndltd.ncl.edu.tw/handle/79058630126022438921.
Повний текст джерелаАнотація:
碩士國立中興大學
獸醫學系
91
The aim of this study is to develop a simple, rapid, and reliable enzyme-linked immunosorbent assay (ELISA) method for detecting the residues of the fluoroquinolones (enrofloxacin, ofloxacin, danofloxacin, and orbifloxacin) in the meats of fish, pork, beef, chicken, and milk. By the N-hydroxysuccinimide ester method, the four tested chemical agents conjugated with BSA, HAS, and HRP had been done, respectively. Successfully, the tested conjugations were able to be detected by the HPLC with UV and fluoresce detector. Moreover, our results shown that the antibody titers by the subcutance immunization way was significantly higher than that intraspleen way in rabbit. Consequently, the titer of the subcuatance immunization was more than 32,768-fold after the fourth booster at four-weeks intervals. The lowest detection limit (LDL) of the four fluoroquinolones (danofloxacin, ofloxacin, orbifloxacin, and enrofloxacin) in PBS, beef, chicken, milk, pork, fish, and FBS were (0.14, 0.60, 1.49, 0.16, 0.61, 0.62, 1.18 ng/mL), (1.18, 3.41, 1.33, 0.22, 3.49, 1.65, 0.27 ng/mL), (0.41, 0.64, 0.70, 0.21, 0.66, 0.78, 0.3 ng/mL), and (2.38, 3.77, 3.61, 3.54, 4.11, 3.59, 4.59 ng/mL) by the ELISA method, respectively. Therefore, the danofloxacin ELISA kit was the most sensitivity in our developed kits. However, it is sufficient sensitivity for the tested fluoroquinolones residues in biological matrices with these kits. The rates of the cross-reaction of the developed danofloxacin, orbifloxacin, ofloxacin, and enrofloxacin ELISA kits to danofloxacin, enrofloxacin, norfloxacin, ofloxacin, orbifloxacin, oxolinic acid, and sarafloxacin were (100%, <0.01%, <0.01%, <0.01%, <0.01%, <0.01%, <0.01%), (<0.01%, <0.01%, 0.01%, <0.01%, 100%, <0.01%, <0.01%), (<0.01%, 0.14%, <0.01%, 100%, <0.01%, <0.01%, <0.01%), and (0.13%, 100%, 0.18%, <0.01%, 0.04%, <0.01%, 0.02%), respectively. Therefore, the above results shown that the best specificity of our developed kit was danofloxacin, then orbifloxacin, ofloxacin, and enrofloxacin. Especially, the antibody of danofloxacin did not have cross-reactivity for the other fluoroquinolones (enrofloxacin, norfloxacin, ofloxacin, orbifloxacin, and sarafloxacin). However, the other antibodies (enrofloxacin, ofloxacin, and orbifloxacin) had low cross-reaction with each other quionlones. To elucidate the variability of standard curves for different assays, standard solutions were prepaerd independently with PBS and analyzed by the ELISA methods. The intra-assay and inter-assay coefficients of variation of danofloxacin, ofloxacin, enrofloxacin, and orbifloxacin ELISA kit were (3.35%, 3.51%), (2.93%, 4.35%), (8.16%, 10.1%), and (7.15%, 9.28%), respectively. Therefore, the best precision of our developed kit was danofloxacin, then ofloxacin, enrofloxacin, and orbifloxacin. From above assay results, our developed ELISA kits shown a simple, rapid, and reproducible method to detect the residues of the tested fluoroquinolones in the meats.
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Wang, Mei-Ying, and 王玫瑛. "Development of enzyme - linked immunosorbent assay ( ELISA ) kit for the diagnosis of Helicobacter pylori infection." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/48779724298183383337.
Повний текст джерелаАнотація:
碩士大仁科技大學
生物科技研究所
94
Helicobacter pylori survives and proliferates in the human gastric mucosa. H pylori infections are usually diagnosed by culture, histology and rapid urease tests of gastric biopsy specimens obtained by endoscopy and noninvasive methods. Urease tests such as enzyme – linked immunosorbent assay ( ELISA ) are commercially available, easy to perform, and inexpensive. Serological diagnoses by ELISA have been widely used in epidemiological studies. The purpose of this study was to use the expressed UreA ( 54 kDa ) and UreB ( 90 kDa ) protein cloned from H. pylori, to evaluate IgA and IgG using ELISA. We compared the IgA and IgG of 40 patients, verified by endoscopy, to determine the presence of H. pylori infection as the gold standard control group. However, totally 157 patients including 40 patients of positive control group were tested. The UreA sensitivity for IgA and IgG were 87 ～ 88 %, and 91 ～ 100 % respectively and the specificity for both IgA and IgG were 100 % and the accuracy for IgA and IgG were 90 ～ 95 %, and 92 ～ 100 % respectively. The UreB sensitivity for IgA and IgG were 87 ～ 88 %, and 94 % respectively and the specificity for IgA and IgG were 95 ～ 100 % and 100 % respectively the accuracy for IgA and IgG were 89 ～ 95 %, and 94 ～ 95 % respectively. The resulting data showed that instead of endoscopy the specific reactivity to IgA or IgG by using ELISA is clinically feasible for the diagnosis of H. pylori infections in humans.
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Wu, Jian-Yu, and 吳建諭. "Development of an enzyme-linked immunosorbent assay (ELISA) kit for the diagnosis of porcine viral diseases by using monoclonal antibody." Thesis, 2012. http://ndltd.ncl.edu.tw/handle/76436343585117854046.
Повний текст джерелаАнотація:
碩士大仁科技大學
生物科技研究所
100
Porcine circovirus type 2 (PCV-2) and porcine reproductive and respiratory syndrome virus (PRRSV) caused the pig sows abortion, weakness of piglets and lymphocyte depletion in lymphoid tissues in infected pigs. These diseases threaten the production of pig and caused huge loss of economics in pig industry. In this study, it focuses on the development of an enzyme-linked immunosorbent assay (ELISA) diagnostic kit by using the purified recombinant protein derived from ORF-7 of PRRSV（rORF-7, 42 kDa）and ORF-2 of PCV-2（rORF-2, 54 kDa）also. An ELISA for detection of the specific antibody against PRRSV by using rORF-7 was carried out very well and the resulting data revealed 100% in sensitivity, 95% in specificity and 97% in accuracy respectively when compared with the commercialized Ingezim PRRS universal ELISA kit produced by Ingenasa Inc. in Spain. Moreover, a monoclonal antibody, 3F12（IgM, κ）, against PRRSV and the specific IgY to PCV-2 were developed and evaluated by using Western blotting assay. The monoclonal antibody 3F12（IgM, κ）and the specific IgY against PCV-2 can be used for the development of an Ag-capture ELISA to detect PRRSV antigens and PCV-2 antigen respectively. The detection limit revealed 1x10-8 TCID50 and 18.5 ng/ml of the purified rORF-7 recombinant protein of PRRSV in terms of the diagnosis of PRRS. By using the specific IgY to PCV-2, an Ag-capture ELISA was also developed and the detection limit showed 39 ng/ml of the purified rORF-2 recombinant protein from PCV-2. The ELISA kits described above could be used for the diagnosis of PRRS or PCV-2. Furthermore, these ELISA kits could also be used for detection of viremia, or evaluation of immune responses after vaccination with PRRSV or PCV-2 vaccines, and for the surveillance of epidemiology of PRRSV or PCV-2 infections in pigs.
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Elji, Rana. "Metodjämförelse mellan två olika enzyme-linked immunosorbent assays (Medizym ICA screen och 2Screen islet cell autoantibody ELISA-kit) för mätning av islet cell antibodies, ICA." Thesis, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:hkr:diva-15298.
Повний текст джерелаАнотація:
Type 1 diabetes (T1D) is regarded as an autoimmune disease. Beta cells, which produces insulin in pancreas are attacked by islet cell antibodies (ICA). This leads to gradual destruction of the beta cell, which in turn cause high level of glucose in the blood because the regulator "insulin" has disappeared. In that case the patient needs to be treated lifelong with insulin. It has been shown that the ICA reactivity consisting of reactivities against different autoantigens such as: insulin autoantigen (IAA), glutamic acid autoantigen (GAD), insulinoma antigen-2 autoantigen (IA-2) and most likely also zinc transporter autoantigen (ZnT8). Determination of ICA in serum samples is important for the classification of diabetes, prediction of T1D and the development of autoimmune therapies. Nowadays screening of ICA is performed with ”Medipan ICA screen” which is a commercial enzyme- linked immunosorbent assay (ELISA). Positives samples are further analysed by ELISA with the indirect immunofluorescence method (IF) to ensure a final positive answer. The purpose of this study was to evaluate and compare a new commercial ELISA kit ”RSR 2screen” with the Medipan ICA screen for use it in routine analysis to evalute if it has the same / higher specificity and sensitivity, and lower price compared with Medipan ICA screen. Serum samples from a control group (n = 199) and a patient’s group diagnosed with T1D (n = 100 were analyzed with both ELISA methods. The results were statistically evaluated to set a threshold value for positivity and to evaluate the method's sensitivity and specificity. The result showed that both ELISA- methods gave the same sensitivity (93%) and specificity (97.5%) and a high concordance (98.7%) was achieved. Analytical price per sample for the RSR 2screen was 4.2% lower than for the Medipan ICA screen. RSR 2screen can be used instead of Medipan ICA screen.
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Yeh, Li-Chun, and 葉俐君. "Development of Amoxicillin Enzyme-Linked Immunosorbent Assay." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/66506324246340310706.
Повний текст джерелаАнотація:
碩士國立中興大學
獸醫學系
92
The residues of β-lactam antibiotics in consumable meat and milk products pose potential risks for public health because of their propensity to induce allergic reactions in individuals and the increasing incidence of microbial resistance against these compounds. Amoxicillin is a β-lactam antibiotics widely used in human and veterinary clinical medicine. The aim of this study was to develop an enzyme -linked immunosorbent assay (ELISA) suitable for detection of amoxicillin in biological fluids. Amoxicillin polyclonal antibody was raised from rabbits immunized with amoxicillin conjugated to bovine serum albumin (BSA) or carboxyl polystyrene particles by either carbodiimide or glutaraldehyde (GA) method. Polystyrene particle was further tested for their efficacy as adjuvant. The results indicated that the highest anti-amoxicillin titer (>102,400 fold dilution) was obtained from rabbits immunized with amoxicillin-BSA conjugates by GA method. Polystyrene particle did not significantly improve the immune response elicit with Freund’s adjuvant. The optimal dilution of amoxicillin antibody for the developed assay was 1,600 fold. The assay has a limit of detection of 1 ng/mL and limit of quantitation of 5 ng/mL in bovine serum, urine and milk. Cross reactivity with penicillin G, ampicillin, oxacillin, cloxacillin, 6-aminopenillinic acid (6-APA), carbenicillin, cefadroxil, cefazolin and dicloxacillin were 77.39％, 56.87％, 51.41％, 48.79％, 37.71％, 36.00％, 7.40％,＜1％ and ＜1％, respectively. The ELISA assay was combined with microdialysis sampling to measure venous and muscle tissue concentrations of amoxicillin in pigeons. After a single intramuscular injection of 25 mg ∕kg amoxicillin, the blood concentration peaked at 4.9 g ∕mL in 20 minutes and declined to about 1 g ∕mL after 6 hours. Amoxicillin concentrations in femoral and breast muscle tissues exhibited a delayed and minor increase compared to those in the blood. The peak concentration in femoral and breast reached 1.1 g ∕mL and 0.7 g ∕mL, respectively, at 60 minutes after drug administration. The protein binding of amoxicillin to pigeon serum was 28 ± 6％. In conclusion, we have developed a sensitive amoxicillin ELISA that could efficiently detect amoxicillin and structurally related β-lactam antibiotics in various biological matrices. The application of this assay in combination with microdialysis has successfully demonstrated its use in tissue pharmacokinetic studies.
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Huang, Hann-shyang, and 黃漢翔. "Development of enzyme-linked immunosorbent assay for dexamethasone." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/17669407357735364037.
Повний текст джерелаАнотація:
碩士國立中興大學
獸醫學系
88
A simple ELISA was developed for the detection of dexamethasone in equine urine, serum and milk. Dexamethasone oxime was prepared by refluxing with -aminooxy acetic acid in ethanol, and was conjugated to bovine serum albumin (BSA) by mixed anhydride method. Anti-dexamethasone serum was obtained after five immunizations. Enzyme conjugate was produced with oxime and horseradish peroxidase (HRP). Checkerboard method was used to determine optimal concentrations of antibodies and enzyme conjugate. A standard curve was established suggesting that it is a competitive immunoassay. Working antiserum dilution ranged from 3200x to 12800x, and 3200x for enzyme conjugate. The detection limit was 1.09ng/mL based on the formula of B/B0 100-2SD%. The detectable range was 1-1000 ng/mL in horse serum and urine, and 1-100 ng/mL in milk. The crossreactivity with flumethasone, betamethasone and triamcinolone was 298%, 9.89% and 5.26%, respectively. The dexamethasone antiserum cross-reacted with 8 natural steroid hormones less than 1%. Intra- and interassay variations were 7.85% and 8.14% in PBS, 8.36% and 8.92% in horse urine, 9.34% and 9.79% in horse serum, 9.23% and 9.91% in raw milk, and 5.20% and 8.57% in fresh milk. A horse was injected via jugular vein with 5mg of dexamethasone phosphate, and blood and urine samples were collected. Urine and serum concentration 30 minutes following dexamethasone administration was 52.75 ng/mL, and 27.42 ng/mL, relatively. Urine and serum levels of dexamethasone remained detectable for 12 and 6 hours, respectively. A dairy cow injected Intramuscularly with 30 mg of dexamethasone phosphate. Milk levels of dexamethasone were 6.8 ng/mL, 6.02 ng/mL, 5.33 ng/mL, 2.19 ng/mL, 1.81 ng/mL, at 1 hour, 4 hours, 12 hours, 24 hours and 36 hours, respectively. Results show that a sensitive and simple ELISA was developed for the detection in horse urine, serum and cow milk without extraction.