酶联免疫试验两步法检测乙型肝炎病毒表面抗原的影响因素分析

摘要

Objective To discuss the influencing factors on two-step enzyme linked immunosorbent assay (ELISA) detection of hepatitis B virus surface antigen (HBsAg ) .Methods Positive ,negative and weakly positive mixed serum were detected under different dilution ,plate washing time ,incubation time and color time .Results With one drop or without dilute solution ,incubation time shortened respectively to 45 and 15 min after adding serum or en-zyme conjugates ,color time changed to 15 min with chromogenic agents ,the weakly positive result was statistically different with standard methods .The positive ,weakly positive and negative results were also statistically different with standard methods when plate was washed after incubation for 1 h .Conclusion The changes of ELISA condi-tions might have obvious effect on weakly positive samples especially those within critical value range (gray zone) , though the effect could be ignored in positive ones .%目的探讨酶联免疫吸附试验（ELISA ）两步法检测乙型肝炎病毒表面抗原（HBsAg ）的影响因素。方法分别在稀释液的量、洗板时机、孵育时间、显色时间等不同条件下对阳性混合血清、阴性混合血清和弱阳性对照血清进行检测。结果使用滴瓶滴加1滴稀释液、不加入稀释液、加入血清后温育时间缩短为45 min 、加入酶结合物后孵育时间缩短为15 min 、加入显色剂后显色时间缩短为15 min ，弱阳性结果较标准方法差异有统计学意义；加入样本孵育1 h 后洗板，阳性、弱阳性和阴性结果均较标准方法差异有统计学意义。结论 ELISA 条件的改变，就强阳性标本而言或许影响不大，但弱阳性标本的结果会被显著影响，尤其是处于临界值（灰区）的标本所受影响可能就更为明显。