蓝耳病病毒抗体ELISA检测试剂盒的“两步法”评价

Evaluation of ELISA Kits for PRRSV Antibody Detection by ‘Two Steps' Method

本研究利用"两步法"对蓝耳病试剂盒进行了使用效果评价:第一步,利用18份参考血清对试剂盒进行初筛,淘汰检测准确率低的试剂盒;第二步,利用参考血清和临床样品,分别开展最低检测限、重复性以及诊断敏感性和诊断特异性分析。结果显示,A、B、C、D 4种市售试剂盒的初筛符合率均为100%,诊断敏感性和诊断特异性分别为94.58%和97.14%、94.58%和89.2%、93.1%和90%、96.55%和85%。其中,A和B为通用型试剂盒,能够检测欧洲型和美洲型蓝耳病;C和D均为美洲型检测试剂盒;重复性上,D试剂盒批内与批间变异系数都较小,重复性较好。本研究首次提出了"两步法"试剂盒评价体系并利用这一体系对4种蓝耳病试剂盒进行了评价,不仅为蓝耳病试剂盒的选择提供了依据,也为今后其它动物疫病检测试剂盒的评价提供了参考。

This study aimed at evaluating four commercial ELISA kits for PRRSV antibody detection by using twosteps for the first time.Firstly,18 reference sera were used to screen.Secondly,6 reference sera were used to detect the limitation of detection（LOD）and repeatability（Re）,and 343 field samples were used to test sensitivity and specificity.The results showed that 4 kits were 100% identical to the reference seras in screening test in the first step,and the sensitivity and specificity of A,B,C and D were 94.58% and 97.14%,94.58% and 89.2%,93.1% and 90%,96.55% and 85% respectively in the second step.Kit A and B are universal kits for detecting antibody against EU and US type of PRRSV.Kit C and D can only detect US type virus antibody.D kit showed coefficient of variation was low among the batches and had good repeatability.The 4 PRRSV kits were varying in detection scale,validity and accuracy.By applying twostep method,this study supplied evidences for the selection of PRRSV antibodies ELISA detection kits,also gave the reference for the evaluation of animal disease detection.