三种ELISA试剂盒检测猪流行性乙型脑炎病毒血清IgG抗体比较

目的 比较国内常用商品化ELISA试剂盒检测猪流行性乙型脑炎病毒血清IgG抗体的诊断效果。方法 选用3种猪乙脑病毒血清IgG抗体ELISA检测试剂盒(Keqian,Lvshiyuan,Tianchen),以微量中和试验为金标准,对90份猪血清进行检测分析。结果 中和试验的阳性检出率为34.4%(31/90),Keqian,Lvshiyuan和Tianchen阳性检出率分别为50.0%(45/90), 47.8%(43/90)和 38.9%(35/90)。灵敏度最高的是Keqian,为90.32%,但其特异度只有71.19%;Tianchen的灵敏度和特异度分别为83.87%和79.66%;Lvshiyuan的灵敏度和特异度分别仅为41.94%和16.95%。与中和试验比较,Tianchen试剂盒的κ系数最高为0.63,其次是Keqian为0.57,Lvshiyuan仅为0.04。显示3种试剂盒具稳定性。结论 目前国内商品化猪乙脑病毒血清IgG抗体ELISA检测试剂盒诊断结果差异较大,与中和试验相比存在较高的假阴性,质量有待提高。

Comparison of three commercial ELISA kits for detection of antibodies against Japanese encephalitis virus in swine sera

The aim of this study was to compare three kinds of commercial enzyme-linked immunosorbent assay (ELISA) kits for detection of antibodies against Japanese encephalitis virus (JEV) in swine sera. A total of 90 swine serum samples were tested simultaneously by three kinds of commercial JEV IgG ELISA kits (Keqian, Lvshiyuan and Tianchen), which were used with the most frequently recorded in domestic literatures. Diagnostic test methods were used for evaluatlon. Neutralization test was used as the gold standard’ test. Results showed that all the three kinds of commercial ELISA kits had higher positive detection rate than that of neutralization test (34.4%, 31/90), the rates of Keqian, Lvshiyuan and Tianchen were 50.0% (45/90), 47.8% (43/90) and 38.9% (35/90), respectively. Both of Lvshiyuan and Tianchen had a result of suspicious positive, the rates were 32.2% (29/90) and 4.4% (4/90) respectively. Sensitivity and specificity of Tianchen was 83.87% and 79.66%, respectively, with the highest positive likelihood ratio (4.12); Keqian had the highest sensitivity (90.32%) and a lower specificity (71.19%), with the highest negative likelihood ratio (0.14); while Lvshiyuan had the lowest sensitivity and specificity, which was 41.94% and 16.95%, respectively. There was no significant difference of detected rates between Tianchen and the gold standard’ serum neutralization test with the highest goodness of fit (κ = 0.63). Comparison results of Keqian and Lvshiyuan with neutralization test were significantly different: the κ coefficient was 0.57 and 0.04, respectively. Additionally, internal consistency analysis suggested that all the ELISA kits had good stability. In conclusion, currently commercially available ELISA kits for detection of JEV IgG antibody in swine sera are quite various. A high false negative was still found in these kits compared with neutralization test. Our results suggest the need to improve the insure of authenticity and reliability in use of commercial JEV IgG ELISA kits.