乙型肝炎表面抗原(HBsAg)酶联免疫吸附试验检测灰区设置的探讨

Discussion on the Setting of Gray Zone for Detection of Hepatitis B Surface Antigen(HBsAg)Enzyme-linked Immunosorbent Assay

目的对乙型肝炎表面抗原(HBsAg)酶联免疫吸附试验检测结果分析,探讨酶联免疫吸附试验(ELISA)灰区设置的意义。方法对2016年1月—2017年6月无偿献血者27214份标本同时采用两个厂家酶联免疫吸附试验(ELISA)试剂检测,该实验室两种试剂均设有灰区,0.5≤S/CO<1,双试剂有反应性,则报告有反应性;单试剂有反应性,双孔复查,复查只要其中一孔有反应性,则报告有反应性,两孔均为无反应性,则报告无反应性。报告为反应性标本。所有检验标本均同时进行HBV-DNA检测。结果共检测27214份血液标本,双试剂有反应性57份,双试剂有反应性中核酸检测有反应性43份;单灰区标本70份,核酸检测均为无反应性;单试剂反应性标本84份,核酸检测均为无反应性;双试剂酶免检测均无反应性标27003份,核酸检测有反应性23份。临界值(Cot off)S/CO为0.5时,A试剂ROC曲线下面积为0.845,95%信置区间为0.719~0.970,B试剂ROC曲线下面积为0.837,95%信置区间为0.725~0.984;临界值S/CO为1时,A试剂ROC曲线下面积为0.792,95%信置区间为0.632~0.953,B试剂ROC曲线下面积为0.792,95%信置区间为0.653~0.930。该实验室试剂A和试剂B最佳临界值(Cut off)为(S/CO值)0.9。结论随着检测技术的提高,乙型肝炎表面抗原(HBsAg)酶联免疫吸附试验(ELISA)检测灰区的设置是否取消,需要进行科学的性能评价,寻找适合该实验室的最佳临界值。

Objective To analyze the detection results of hepatitis B surface antigen(HBsAg)enzyme-linked immunosorbent assay,and to explore the significance of the gray zone setting of the enzyme-linked immunosorbent assay(ELISA).Methods 27214 samples of unpaid blood donors from January 2016 to June 2017 were tested with enzyme-linked immunosorbent test(ELISA)reagents from two manufacturers at the same time.Both reagents in the laboratory had gray areas,0.5≤S/CO<1,if the two reagents were reactive,the report had reactivity;the single reagent had reactivity,double-hole recheck,recheck as long as one of the holes was reactive,then the report was reactive,and both holes had no reactivity.Reported as a reactive specimen.All test specimens were tested for HBV-DNA at the same time.Results A total of 27,214 blood samples were tested,57 were reactive with dual reagents,43 were reactive with nucleic acid detection in dual reagents,and 70 samples from single gray zone were non-reactive in nucleic acid detection;single reagents were reactive.There were 84 specimens,all of which were non-reactive in nucleic acid detection;27,003 samples were non-reactive in double reagent enzyme immunoassay,and 23 were reactive in nucleic acid detection.When the cutoff value(Cot off)S/CO was 0.5,the area under the ROC curve of reagent A was 0.845,the 95%confidence interval was 0.719-0.970,the area under the ROC curve of reagent B was 0.837,and the 95%confidence interval was 0.725-0.984;When the critical value S/CO was 1,the area under the ROC curve of reagent A was 0.792,and the 95%confidence interval was 0.632-0.953,the area under the ROC curve of reagent B was 0.792,and the 95%confidence interval was 0.653-0.930.The best cut off of reagent A and reagent B in this laboratory was(S/CO value)0.9.Conclusion With the improvement of detection technology,whether the setting of the gray area for detection of hepatitis B surface antigen(HBsAg)enzyme-linked immunosorbent assay(ELISA)is cancelled,scientific performance evaluation is needed to find the best cutoff value suitable for our laboratory.