酶联免疫法检测血清乙型肝炎表面抗原的空白限检出限及定量限的建立与评价

目的探讨酶联免疫吸附试验(ELISA)检测血清乙型肝炎表面抗原(HBsAg)结果检测限的问题,防止漏检弱阳性标本,建立本实验室的空白限(LOB)、检出限(LOD)及定量限(LOQ)。方法参考美国临床实验室标准化委员会(CLSI)发布的《确定检测低限和定量检测限的方案》(EP17-A)文件,将HBsAg的空白样本及一系列的低浓度样本进行ELISA检测,对得到的结果根据其分布状态,采用百分位数进行统计描述,建立ELISA法检测HBsAg的空白限、检出限及定量限。结果空白限为0.117μg/L,检出限为0.317μg/L,定量限为0.500μg/L。结论参考CLSIEP17-A文件建立的检测限比常规方法建立的检测限科学合理,更能满足临床实验室的要求。

The establishment and evaluation of limit of blank, limit of detection and limit of quantitation for detection of serum hepatitis B surface antigens by enzyme-linked immunosorbent assay

Objective To explore the question of serum hepatitis B surface antigens（HBsAg） results detected by enzyme-linked immunosorbent assay（ELISA） so as to prevent undetection of weakly positive samples, and to estab- lish limit of blank （LOB）, limit of detection （LOD） and limit of quantitation （LOQ）. Methods Refering to CLSI EP17-A file, the HBsAg blank sample and a series of samples with low concentrations were performed ELISA detection. According to the distribution, the resuhs were analyzed using the relative statistical method. ELISA was used to detect LOB, LOD and LOQ of HBsAg. Results The results were LOB=0.117μg/L, LOD=0.317μg/L and LOQ= 0.500μg/L. Conclusion The LOB established by referring to CLSI EP17-file was more scientific and reasonable than that established by conventional method and thus it might satisfy the clinical lab requirements better.