Afshar A, Shakarchi N H, Dulac G C
Animal Diseases Research Institute, Agriculture Canada, Nepean, Ontario.
J Clin Microbiol. 1993 Jul;31(7):1860-5. doi: 10.1128/jcm.31.7.1860-1865.1993.
Two competitive (C) enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of antibodies to vesicular stomatitis virus (VSV) in animal sera. The assays are based upon the availability of polyclonal antibodies (PAbs) from mouse ascitic fluids prepared against the New Jersey (NJ) and the Indiana (IN) VSV serotypes. The assays were performed by the immobilization of VSV-NJ and VSV-IN antigens on a solid phase (microtiter plate). Appropriately diluted test serum mixed with an equal volume of serotype-specific PAb was allowed to incubate in the presence of the relevant VSV antigen and finally with an enzyme-conjugated anti-mouse immunoglobulin. In the absence of anti-VSV antibodies in the test serum, the VSV antigen sites are reactive with the relevant PAb (NJ or IN) as indicated by color development after enzyme degradation of substrate. If the test serum contains the homologous VSV-NJ or VSV-IN antibodies, they compete with the relevant PAb for immobilized antigen sites and quantitatively block and inhibit the PAb reaction and subsequent color development. The performance of C-ELISAs in detecting anti-VSV antibodies in serum samples from four calves, two horses, four sheep, and seven pigs experimentally infected with VSV-NJ and VSV-IN was evaluated. The sensitivity and specificity of the C-ELISAs were compared with those of the standard microtiter serum neutralization (MTSN) tests. Homologous antibodies were demonstrable by C-ELISAs as early as 5 days postinfection (DPI) in one horse and one sheep infected with VSV-IN serotype. Seroconversion was demonstrable by C-ELISAs and MTSN tests in all animals by 9 DPI except in one sheep that received VSV-NJ and one horse inoculated with VSV-IN serotype which, on the basis of the MTSN test results, did not seroconvert until 14 and 11 DPI, respectively. The dynamics of homologous antibody response in all animals as revealed by the corresponding type-specific C-ELISAs paralleled the results of the MTSN tests. The type-specific antibodies to VSV serotypes increased exponentially during the first 2 to 4 weeks postinfection and remained relatively stable for about 6 months in some animals. The results suggest that the C-ELISAs offer many advantages over the MTSN tests and have potential applications as rapid and inexpensive tests in serodiagnosis of VSV infections in animals.
开发了两种竞争性（C）酶联免疫吸附测定法（ELISA），用于检测动物血清中针对水疱性口炎病毒（VSV）的抗体。这些测定法基于从针对新泽西州（NJ）和印第安纳州（IN）VSV血清型制备的小鼠腹水获得的多克隆抗体（PAbs）。通过将VSV-NJ和VSV-IN抗原固定在固相（微量滴定板）上来进行测定。将适当稀释的测试血清与等体积的血清型特异性PAb混合，在相关VSV抗原存在下孵育，最后与酶标记的抗小鼠免疫球蛋白孵育。如果测试血清中不存在抗VSV抗体，则VSV抗原位点与相关PAb（NJ或IN）反应，底物经酶降解后显色即表明这一点。如果测试血清含有同源的VSV-NJ或VSV-IN抗体，它们会与相关PAb竞争固定的抗原位点，并定量阻断和抑制PAb反应及随后的显色。评估了C-ELISA在检测实验感染VSV-NJ和VSV-IN的四头小牛、两匹马、四只绵羊和七头猪的血清样本中抗VSV抗体的性能。将C-ELISA的敏感性和特异性与标准微量滴定血清中和（MTSN）试验的敏感性和特异性进行了比较。在感染VSV-IN血清型的一匹马和一只绵羊中，早在感染后5天（DPI），C-ELISA就可检测到同源抗体。除了一只接种VSV-NJ的绵羊和一匹接种VSV-IN血清型的马外，在所有动物中，到9 DPI时，C-ELISA和MTSN试验均可检测到血清转化，根据MTSN试验结果，这只绵羊和这匹马分别直到14 DPI和ll DPI才发生血清转化。相应的型特异性C-ELISA所揭示的所有动物中同源抗体反应的动态与MTSN试验结果相似。VSV血清型的型特异性抗体在感染后的前2至4周呈指数增加，在一些动物中约6个月保持相对稳定。结果表明，C-ELISA比MTSN试验具有许多优势，并且作为动物VSV感染血清学诊断中的快速且廉价的检测方法具有潜在应用价值。
