黄利君 常凡 莫展

[摘要] 目的 驗证时间分辨荧光分析法检测乙肝表面抗原（HBsAg）的检出限（LOD），评价参考区间的合适性。 方法 自2017年10月～2018年1月对HBsAg二级标准物倍比稀释后采用时间分辨荧光分析法及酶联免疫吸附法（ELISA）进行配对检测，标准物浓度除以稀释后检测结果出现阴性的上一稀释度为LOD，验证厂商的声明，比较两种方法检测同一稀释度样品结果，评估厂商声明的参考区间的合适性;根据美国临床实验室标准委员会（CLSI）EP17-A2规定的方法建立空白限（LOB）及LOD，评价已建立的LOD作为参考区间上限的合适性。 结果 时间分辨荧光法的LOD验证结果为0.1 U/mL，与厂商声明相符。如使用时间分辨荧光检测系统厂商声明的参考区间，标准物稀释3倍后结果判为假阴性，与ELISA检测结果不符。重庆医科大学附属永川医院（以下简称“我院”）检验科建立的HBsAg的LOB及LOD分别为0.048 ng/mL及0.137 ng/mL，如以我院检验科建立的LOD制订参考区间，稀释3倍后两种方法结果一致，均为阳性。 结论 时间分辨免疫荧光分析系统声明的LOD可以接受，但厂商声明的参考区间不适宜，易导致假阴性。检验科应建立自己的LOD来制订参考区间。建议在对病毒标志物定量检测项目进行性能验证时增加参考区间的验证。

[关键词] 乙肝表面抗原;EasyCuta全自动时间分辨免疫荧光分析;LOD;参考区间

[中图分类号] R446.1          [文献标识码] A          [文章编号] 1673-7210（2019）03（a）-0149-04

[Abstract] Objectives To verify the detection limit （LOD） of hepatitis B surface antigen （HBsAg） by time-resolved fluorimmunoassay， and to evaluate the suitability of the reference interval. Methods From October 2017 to January 2018， the HBsAg secondary standard substance was diluted and then paired with time-resolved fluorescence method and enzyme-linked immunosorbent assay （ELISA） for detection， the detection limit was the last dilution degree with negative test results after standard substance concentration divided by dilution， to verify the manufacturer′s statement， the results of samples with the same dilution degree detected by the two methods was compared， and the suitability of the reference interval declared by the manufacturer was evaluated. According to the American clinical laboratory standards board （CLSI） EP17-A2 method， create blank limits （LOB） and LOD， and the suitability of the established LOD as the upper limit of the reference interval was evaluated. Results The detection limit of time-resolved fluorimmunoassay method was 0.1 U/mL， which was consistent with the manufacturer′s statement. If the reference interval declared by the manufacturer of the time-resolved fluorimmunoassay detection system was used， the standard substance was diluted 3 times and the result was found to be false negative， which was inconsistent with the ELISA test result， the LOB and LOD of HBsAg established in Department Clinical Laboratory， Yongchuan Hospital of Chongqing Medical University （hereinafter heferred to as "our hospital"）， were 0.048 ng/mL and 0.137 ng/mL， respectively. If the reference interval was established with the LOD established chinical laboratory of our hospieal， the results of the two methods were consistent after 3 times of dilution， and both were positive. Conclusion The detection limit declared by the time-resolved fluorimmunoassay analysis system is acceptable， but the reference interval declared by the manufacturer is not appropriate， which is likely to lead to false negative. The clinical laboratory should establish its own LOD to establish the reference interval. It is suggested to increase the validation of the reference interval in the performance verification of the quantitative detection items of virus markers.

[Key words] Hepatitis B surface antigen; EasyCuta automatic time-resolved fluorescence immunoassay; Limit of detection; Reference interval

目前定量检测病毒标志物的方法（如时间分辨荧光分析法及化学发光方法）虽声称是一个定量实验，但其本质就是一个通过参考区间对定量结果进行阴阳性判断的定性实验，其声明的参考区间的上限其实就是检出限（LOD）。……