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间接酶联免疫吸附测定法在检测牛抗水疱性口炎病毒抗体中的应用。

Application of indirect ELISA for detection of bovine antibodies against vesicular stomatitis viruses.

作者信息

Afshar A, Dulac G C, Wright P F, Martin D

机构信息

Animal Diseases Research Institute, Agriculture Canada, Nepean, ON.

出版信息

J Vet Diagn Invest. 1993 Jan;5(1):26-32. doi: 10.1177/104063879300500107.

Abstract

Two indirect enzyme-linked immunosorbent assays (I-ELISAs) are described for the detection of bovine serum antibody to the New Jersey (NJ) and Indiana (IN) vesicular stomatitis viruses (VSV). Serum samples at a dilution of 1:200 were incubated with binary ethylenimine-inactivated VSV-NJ and VSV-IN type-specific antigens preadsorbed to microtiter plates. Bound antibodies were detected by a murine monoclonal antibody to bovine IgG1 conjugated with horseradish peroxidase. The performance of each I-ELISA in detecting homotypic and heterotypic antibodies to VSV-NJ and VSV-IN in sequential serum samples from calves experimentally infected with VSV-NJ or VSV-IN was evaluated. The I-ELISAs detected serotype-specific antibodies to either VSV-NJ or VSV-IN in calves infected with the homologous serotype. Homotypic but not heterotypic anti-VSV-NJ antibodies were first demonstrable by the VSV-NJ I-ELISA during the second week postinfection and remained at an elevated level for a period of 11 weeks, with a gradual decrease thereafter. Similar homotypic antibody profiles measured by the VSV-IN I-ELISA in calves inoculated with VSV-IN were observed. The performances of the I-ELISAs were compared using 1,495 microtiter serum neutralization (MTSN) test-negative bovine field sera collected from cattle in Canada (VS free) and 429 samples collected from cattle in the USA and Mexico (VS-epidemic and VS-endemic areas). The diagnostic specificities of the VSV-NJ and VSV-IN I-ELISAs for the Canadian samples relative to the MTSN test results were in the range of 99.8% and 99.7%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

本文描述了两种间接酶联免疫吸附测定法(I-ELISA),用于检测牛血清中针对新泽西(NJ)和印第安纳(IN)型水疱性口炎病毒(VSV)的抗体。将稀释至1:200的血清样本与预先吸附在微量滴定板上的经双乙烯亚胺灭活的VSV-NJ和VSV-IN型特异性抗原一起孵育。通过与辣根过氧化物酶偶联的抗牛IgG1鼠单克隆抗体检测结合的抗体。评估了每种I-ELISA在检测实验感染VSV-NJ或VSV-IN的犊牛连续血清样本中针对VSV-NJ和VSV-IN的同型和异型抗体方面的性能。I-ELISA在感染同源血清型的犊牛中检测到针对VSV-NJ或VSV-IN的血清型特异性抗体。在感染后第二周,VSV-NJ I-ELISA首次检测到同型而非异型抗VSV-NJ抗体,其水平在11周内一直升高,此后逐渐下降。在接种VSV-IN的犊牛中,通过VSV-IN I-ELISA观察到了类似的同型抗体谱。使用从加拿大(无VS)的牛采集的1495份微量滴定血清中和(MTSN)试验阴性的牛场血清以及从美国和墨西哥(VS流行和VS地方性流行地区)的牛采集的429份样本,比较了I-ELISA的性能。相对于MTSN试验结果,VSV-NJ和VSV-IN I-ELISA对加拿大样本的诊断特异性分别在99.8%和99.7%的范围内。(摘要截短至250字)

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