HEV抗体检测中ELISA试剂盒的性能验证研究

Study on performance verification of ELISA kit for HEV antibody detection

目的探讨酶联免疫吸附试验(ELISA)检测中试剂盒的性能验证方法,并对该实验室戊型肝炎病毒(HEV)抗体检测中所使用的ELISA试剂盒进行性能验证,以判断所用试剂盒是否能满足实验室检测工作的基本要求。方法使用FAME全自动酶联免疫分析仪进行ELISA检测,通过计算和分析所用试剂盒的最低检出限、重复性、期间精密度、正确度,与试剂盒说明书提供的性能指标进行对比,并对试剂盒的CUTOFF值进行验证,判断其是否适合实验室常规检测人群,以此评价试剂盒的性能指标。结果 HEV ELISA检测中,万泰试剂盒最低检出限为0.5 U/m L,GBI试剂盒最低检出限为1 U/m L;重复性精密度实验中,万泰试剂盒在弱阳性浓度水平(2 U/m L)和临界值浓度水平的变异系数(CV)分别为4.33%和5.84%,GBI试剂盒在弱阳性浓度水平(2 U/m L)和临界值浓度水平的CV分别为5.39%和7.82%;期间精密度实验中,万泰试剂盒在弱阳性浓度水平(2 U/m L)和临界值浓度水平的CV分别为8.89%和12.55%,GBI试剂盒在弱阳性浓度水平(2 U/m L)和临界值浓度水平的CV分别为8.57%和12.52%;正确度验证方面,2种试剂盒的阴阳性符合率都达到100%;CUTOFF值验证实验中,万泰试剂盒检测的样本OD值的+3SD=0.190,小于试剂盒提供的CUTOFF值0.263,GBI试剂盒检测的样本OD值的+3SD=0.074,小于试剂盒提供的CUTOFF值0.150。结论实验室所使用的2种试剂盒均通过性能验证,适合实验室日常检测工作和受检人群,对提高实验室检测的准确度和检测质量起到了积极作用。

Objective To investigate the performance verification method of the ELISA kit,and to perform the performance verification of the ELISA kit for hepatitis E virus（HEV） antibody detection to judge whether the used kit could meet the basic requirements of laboratory detection work. Methods The FAME fully automatic enzyme-linked immunity analyzer was used to conduct the ELISA detection,the lowest limit of detection,repeatability,intermediate precision and accuracy by calculation and analysis were compared with the performance indexes provided by the kit instruction,and the cut off value of kit was verified for judging whether this kit being suitable for the detection of laboratory routine population,thus for evaluating the performance indexes of the kit. Results In the ELISA detection for HEV,the lowest detection limit of Wantai kit was 0.5 U/m L,which of GBI kit was 1 U/m L;in the precision tests of repeatability,CV of Wantai kit was 4.33% at the weakly positive concentration level（2 U/m L） and 5.84% at the critical value concentration level,which of GBI kit was 5.39% at the weakly positive concentration level（2 U/m L） and 7.82% at the critical value concentration level;in the intermediate precision test,CV of WANTAI kit was 8.89% at the weakly positive concentration level and 12.55% at the critical value concentration level,which of GBI kit was 8.57% at the weakly positive concentration level（2 U/m L） and 12.52% at the critical value concentration level;in the accuracy test,the negative and positive coincidence rates of both kits all reached 100%;in the cutoff value verification test,x＋3SD=0.190 in the OD value of WANTAI kit sample,which was less than the cutoff value 0.263 provided by the kits,and x＋3SD=0.074 in the OD value of GBI kit sample was 0.074,which was less than the cutoff value 0.150 provided by the kit. Conclusion Both ELISA kits used by the laboratory pass the performance verification,are suitable for the routine detection and detected groups,which plays an active role for increasing accuracy and quality of laboratory detection.