College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
College of Liberal Arts and Sciences, Iowa State University, Ames, Iowa, USA.
mSphere. 2019 Mar 13;4(2):e00017-19. doi: 10.1128/mSphere.00017-19.
This study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples ( = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue ( = 12), TGEV strain Miller ( = 12), PRCV ( = 12), or with virus-free culture medium ( = 12). ELISA results were evaluated both with "suspect" results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (0.05) cross-reactivity between TGEV and PRCV serum antibodies, particularly during early stages of the infections. Serologic cross-reactivity between TGEV and PRCV seemed to be TGEV strain dependent, with a higher percentage of PRCV-false-positive results for pigs inoculated with TGEV Purdue than for TGEV Miller. Moreover, the overall proportion of false positives was higher when suspect results were interpreted as positive, regardless of the ELISA kit evaluated. Current measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals. However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis.
本研究使用血清样本( = 528)比较了三种商业传染性胃肠炎病毒/猪呼吸道冠状病毒(TGEV/PRCV)阻断酶联免疫吸附试验(ELISA)的性能,这些样本是在接种 TGEV 菌株 Purdue( = 12)、TGEV 菌株 Miller( = 12)、PRCV( = 12)或无病毒培养基( = 12)的猪在 49 天观察期内采集的。ELISA 结果分别用“疑似”结果解释为阳性和阴性进行了评估。当检测来自接种无病毒培养基的猪的样本时,所有商业试剂盒均显示出优异的诊断特异性(99%至 100%)。然而,分析表明试剂盒在诊断敏感性(TGEV 或 PRCV 血清抗体阳性的猪百分比)方面存在差异,所有试剂盒均显示 TGEV 和 PRCV 血清抗体之间存在显著(0.05)交叉反应,特别是在感染的早期阶段。TGEV 和 PRCV 之间的血清交叉反应似乎取决于 TGEV 株,与接种 TGEV 米勒的猪相比,接种 TGEV 普渡的猪的 PRCV 假阳性结果比例更高。此外,无论评估哪种 ELISA 试剂盒,当将疑似结果解释为阳性时,假阳性的总体比例更高。目前预防 TGEV 进入无疫区的措施包括检疫和检测 TGEV 血清阴性动物。然而,由于 PRCV 的交叉反应,TGEV 血清学变得复杂,PRCV 在世界上大多数猪群中呈亚临床传播。常规血清学检测不能区分 TGEV 和 PRCV 抗体;然而,使用含有 PRCV S 蛋白氨基末端大缺失的抗原的阻断 ELISA 可以区分 PRCV 和 TGEV 抗体。有几种商业 TGEV/PRCV 阻断 ELISA 可用,但最近的研究没有报告性能比较。本研究表明,TGEV 和 PRCV 之间的血清交叉反应会影响商业阻断 ELISA 的准确性。必须谨慎解释个别检测结果,尤其是在出现疑似结果的情况下。因此,商业 TGEV/PRCV 阻断 ELISA 仅应在群体基础上应用。