Abstract: Objective　Evaluate the performance of various mouse hepatitis virus (MHV) enzyme-linked immunosorbent assay (ELISA) kits, and compare the concordance rate of the detection result between ELISA and quantitative reverse transcription polymerase chain reaction (RT-qPCR) method.Methods In this experiment, 179 clinical serum samples of mice were used to compare the specificity, sensitivity, inter-batch and intra-batch differences, compliance rate and other performance differences between five Elisa kits and B kits. RT-qPCR method was used to detect 91 mice samples from tracheal+lung, liver and fecal samples, and the result were compared with the result of Elisa.Results　Using the test result of the B kit as A reference standard, the result showed that the A kit showed false positives in the negative samples and the C kit showed false negatives in the positive samples. The highest dilution gradient that the A and C Elisa kits could detect positive serum was both 2-5. The intra-batch and inter-batch differences of A kit were kept within 15%, and the intra-batch differences of C kit were kept within 15%, while the inter-batch differences significantly exceeded 15%. Compared with B kit, the positive coincidence rate of A kit was 96%, the negative coincidence rate was 98%, and the comprehensive coincidence rate was 97%. Compared with B kit, C kit had a positive coincidence rate of 93%, a negative coincidence rate of 92%, and a comprehensive coincidence rate of 92%. Secondly, the coincidence rate of MHV detection by Elisa and RT-qPCR was 69%, and the MHV positive result detected by RT-qPCR were all detected in fecal samples. Rest 3 kits did not show positive result. Conclusion　With the test result of B kit as the reference standard, Among the five MHV Elisa test kits, kit A had the best comprehensive performance, such as specificity, sensitivity, difference between and within batches and coincidence rate. The coincidence rate of MHV detection by Elisa and RT-qPCR was 69%. Rest 4 kits did not show positive result accurately.

Key words: MHV, Elisa kit, RT-qPCR, comparison test

摘要： 目的　评估多种小鼠肝炎病毒(MHV)酶联免疫吸附试验(Elisa)检测试剂盒的性能,并对比Elisa与定量反 转录聚合酶链反应(RT-qPCR)两种方法检测结果的符合率。方法　本实验采用179份临床小鼠血清,比较5种 Elisa试剂盒与B试剂盒的特异性、敏感性、批间和批内差异、符合率等性能差异。并利用RT-qPCR方法检测91份 小鼠气管+肺、肝和粪便样品,对比Elisa检测的结果。结果　以B试剂盒的检测结果作为参考标准,结果显示,A 试剂盒在阴性样品中检测出假阳性,C试剂盒在阳性样品中检出假阴性。A和C两种MHV Elisa试剂盒可检出阳 性血清的最高稀释梯度均为2-5。A试剂盒的批内和批间差异均保持在15%以内;C试剂盒的批内差异均保持在 15%以内,而批间差异明显超出了15%。A试剂盒与B试剂盒相比,阳性符合率为96%,阴性符合率为98%,综合 符合率为97%;C试剂盒与B试剂盒相比,阳性符合率为93%,阴性符合率为92%,综合符合率为92%。其次,Elisa 和RT-qPCR方法的MHV检测结果符合率为69%,且RT-qPCR方法检出的MHV阳性均是在粪便样品中检出。其 余3种试剂盒未显示阳性结果。结论　以B试剂盒的检测结果作为参考标准,5种MHV Elisa检测试剂盒中A试 剂盒的特异性、敏感性、批间和批内差异、符合率等综合性能最优;Elisa和RT-qPCR的MHV检测结果符合率为 69%。其余4种试剂盒未能准确显示阳性结果。

关键词: MHV, Elisa试剂盒, RT-qPCR, 比对实验