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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus
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Validation of a Blocking Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus

机译：用于检测针对猪繁殖与呼吸综合征病毒的抗体的封闭酶联免疫吸附法的验证

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Porcine reproductive and respiratory syndrome (PRRS) continues to be one of the most significant diseases of swine. IDEXX HerdChek PRRS, a commercially available enzyme-linked immunosorbent assay (ELISA), has become the industry standard for the detection of antibodies against PRRS virus (PRRSV). The need to accurately determine the PRRSV serostatus of herds and individual animals has prompted the development of several follow-up assay methods. A highly specific and repeatable blocking ELISA (bELISA) was developed on the basis of the use of an expressed PRRSV nucleocapsid (N) protein as the antigen and a biotinylated monoclonal antibody. Validation of the bELISA used sera from 316 animals experimentally and naturally infected with North American PRRSV and sera from 370 uninfected animals. Receiver operating characteristic analysis of the data calculated a diagnostic sensitivity of 97.8% and a diagnostic specificity of 100%. The between-run coefficient of variation of an internal quality control serum was 4.24%. The bELISA was able to detect seroconversion as well as the IDEXX ELISA and the indirect fluorescent antibody (IFA) assay; kappa values were 0.94 and 0.96, respectively. A collection of 133 serum samples with unexpected positive IDEXX ELISA results was obtained from 4,038 diagnostic samples submitted from farms from which PRRS-negative results were expected. The bELISA identified 97% of the samples as PRRS seronegative, while the IFA identified 100% as seronegative. The anticipated use of the bELISA is as a follow-up test to the IDEXX ELISA for determining the PRRSV serostatus of individual animals with unexpected positive test results from swine herds from which negative results are expected.
机译：猪繁殖与呼吸综合症（PRRS）仍然是猪中最重要的疾病之一。 IDEXX HerdChek PRRS，一种市售的酶联免疫吸附测定（ELISA），已成为检测PRRS病毒（PRRSV）抗体的行业标准。准确确定牛群和个别动物的PRRSV血清状况的需求促使了几种后续检测方法的发展。在使用表达的PRRSV核衣壳蛋白（N）作为抗原和生物素化单克隆抗体的基础上，开发了高度特异性和可重复的封闭ELISA（bELISA）。通过实验验证了316种动物的bELISA血清的有效性，并通过自然感染了北美PRRSV和370种未感染动物的血清进行了自然感染。数据的接收者工作特征分析计算出诊断灵敏度为97.8％，诊断特异性为100％。内部质量控制血清的批间变异系数为4.24％。 bELISA能够检测血清转化，IDEXX ELISA和间接荧光抗体（IFA）分析。卡伯值分别为0.94和0.96。从从农场提交的4,038个诊断样本中获得了133个血清的IDEXX ELISA阳性结果，这些样本具有令人惊讶的PRRS阴性结果。 bELISA将97％的样品鉴定为PRRS血清阴性，而IFA将100％的样品鉴定为血清阴性。 bELISA的预期用途是作为IDEXX ELISA的后续测试，用于确定单个动物的PRRSV血清状况，猪群具有出乎意料的阳性测试结果，而预期的阴性结果。

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