Clavijo A, Zhou E M, Vydelingum S, Heckert R
National Center for Foreign Animal Disease, Winnipeg, Manitoba, Canada.
Vet Microbiol. 1998 Feb 28;60(2-4):155-68. doi: 10.1016/s0378-1135(98)00160-6.
An antigen-capture enzyme immunoassay (EIA) was developed to detect classical swine fever virus (CSFV) antigen directly from 10% w/v tissue suspension. The assay, based on the sandwich principle, uses a biotinylated monoclonal antibody bound to streptavidin-coated microplates as the capture system and a swine anti-CSFV antibody and rabbit anti-swine HRPO-conjugate as the detector system. The antigen-capture EIA was compared with conventional virus isolation and polymerase chain reaction (PCR) for detection of CSFV in tissues. The ability of the antigen-capture EIA to discriminate classical swine fever (CSF) from bovine viral diarrhea and African swine fever viruses was also tested. The assay was shown to detect 21 different strains of CSFV and was unreactive with tissues from uninfected animals. Signal to noise (S/N) ratios were calculated from the EIA absorbance values. Readings from samples positive by virus isolation (n = 47) averaged a S/N ratio of 5.34. In contrast, samples negative by virus isolation (n = 96) demonstrated a mean S/N ratio of 0.16. At S/N cut-off value of 1.0, all samples that yield virus isolation and PCR negative result were negative in the antigen-capture EIA. Compared with virus propagation in tissue culture using PK15 cells (followed by indirect peroxidase assay detection) and PCR, the EIA had a specificity of 98.7% and a sensitivity of 91.4%. The EIA is simple, can be performed in 4 h and lends itself to automation for screening of tissues sample from pigs suspected of CSFV infection.
开发了一种抗原捕获酶免疫测定法（EIA），用于直接从10%（w/v）组织悬液中检测经典猪瘟病毒（CSFV）抗原。该测定法基于三明治原理，使用与链霉亲和素包被的微孔板结合的生物素化单克隆抗体作为捕获系统，以及猪抗CSFV抗体和兔抗猪HRPO缀合物作为检测系统。将抗原捕获EIA与传统病毒分离法和聚合酶链反应（PCR）进行比较，以检测组织中的CSFV。还测试了抗原捕获EIA区分经典猪瘟（CSF）与牛病毒性腹泻和非洲猪瘟病毒的能力。结果表明，该测定法可检测21种不同的CSFV毒株，且与未感染动物的组织无反应。根据EIA吸光度值计算信噪比（S/N）。病毒分离呈阳性的样本（n = 47）的读数平均信噪比为5.34。相比之下，病毒分离呈阴性的样本（n = 96）的平均信噪比为0.16。在S/N截止值为1.0时，所有病毒分离和PCR结果为阴性的样本在抗原捕获EIA中均为阴性。与使用PK15细胞在组织培养中进行病毒增殖（随后通过间接过氧化物酶测定法检测）和PCR相比，EIA的特异性为98.7%，灵敏度为91.4%。该EIA操作简单，4小时即可完成，适合自动化操作，用于筛查疑似感染CSFV的猪的组织样本。
