ELISA法检测乙肝标志物影响因素的探讨

目的 探讨酶联免疫吸附试验（ELISA）检测乙肝血清标志物的影响因素。方法 用ELISA法检测5项乙肝血清标志物，对影响检测结果的几个重要因素进行统计分析。结果 5项乙肝标志物的孔间差变异系数（CV）为5％～25％，常规质控CV为18％～65％，阳性／阴性对照CV为4％～31%。用ELISA法和微粒子酶免分析（MEIA）法分别检测抗HBc，其结果有极显著性差异（p＜0.01）。在HBBAg、抗HBe检测中周围孔和中央孔吸光度（A）值比较，P＜0．01和P〈0．05。终止反应后随比色时间的延长A值逐渐下降，并且HBeAg浓度越高其A值下降越快。不同稀释方法检测抗HPc，各组间A值比较差异有统计学意义。随样品与加入酶结合物间隔时间的延长，假阳性率逐渐增加。结论 ELISA检测结果的可靠性主要受试剂质量（尤其是抗HBe、抗HBc）和检测过程中操作误差的影响；质控物浓度的高低可以影响常规质控CV的大小。

Study on the influential factors of HBV markers detection by ELISA

Objective To analyze the influential factors of HBV markers detection by ELISA. Methods ELISA was used to detect the five HBV markers. The several important factors which influence the results were analyzed. Results The variance between reaction orifices of the five HBV markers was 5%-25%, the routine condition variance was 18%-65% and the positive/negative control variance was 4%-31%. The results of anti-HBc detected by ELISA and MEIA were different,P< 0.01 . In the detection of HBsAg and anti-HBc, the marginal results were not identical with the central results,P< 0.01 ,P< 0.05 , respectively. After adding the stopping liquid, the absorbency values gradually decreased in a time-dependent-manner.The different dilution methods had significant influence on the anti-HBc results. As the interval between the sampling and the adding of conjugates protracted, the false positive rate increased. Conclusion The reliability of the ELISA detection of HBV markers mainly depends on the reagent quality and the operational error in the detection; the control concentration may influence the value of routine condition variance.