分批加酶试剂法消除操作时差对竞争法检测抗-HBc的假阳性影响

Avoiding Fault Positive of Anti-HBc Detection Caused by Operation Time Difference with Plusing Enzyme Reagent in Turn

目的 探讨操作时差对竞争法检测抗-HBc的假阳性影响及消除办法,提高检验质量.方法 方法1：在竞争法检测抗-HBc酶标孔中加入血清标本后放置5,10,15,20,30 min再加酶试剂,然后进行下一步操作;方法2：同时加入血清标本及酶试剂后放置5,10,15,20,30 min,然后再进行下一步操作;方法3：同时加入血清标本及酶试剂,立即进行下一步操作（放置0 min,作为标准对照组）.用上述三种方法对36例抗-HBc阴性标本及30例抗-HBc阳性标本进行实验,对实验结果进行统计分析,探讨操作时差对竞争法检测抗-HBc假阳性的影响及消除办法.结果 方法1：36例抗-HBc阴性标本5,10,15,20,30 min各组吸光度低于对照组,假阳性率分别为16.7%,27.8%,33.3%,36.1%,38.9%;30例抗-HBc阳性标本5,10,15,20,30 min各组吸光度低于对照组,但未造成结果的假阳性及假阴性.方法 2：36例抗-HBc阴性标本及30例抗-HBc阳性标本5,10,15,20,30 min各时间段组与对照组比较吸光度差异均无统计学显著性意义,无一例出现假阳性或假阴性.结论 同时加入血清标本及酶试剂,放置不同时间,将血清标本与酶试剂的不公平竞争时差转变为公平竞争时差,可消除操作时差对竞争法检测抗-HBc的假阳性影响.工作中可在加完一批样本即加入酶试剂,然后加下批标本及酶试剂,即分批加酶试剂法,可保证检测结果的准确性.

Objective To survey fault positive influence of operation time difference to anti-HBc detection and avoidance. Methods Method 1：Plus sample and place them 5,10, 15, 20 and 30 minutes in room temperature then plus enzyme reagent ,next operation was fallowed;Method 2：Plus sample and enzyme reagent immediately and place them 5,10,15,20 and 30 minutes in room temperature,and next operation was fallowed ;Method 3 ：Plus sample and enzyme reagent immediately and do next operation immediately （set as control）. Compare their absorbency and fault positive ratio. Results Method 1 ,absorbency drop obviously and fault positive ratio rise in 36 cases of anti-HBe negative samples ,while in 30 anti-HBc positive samples,absorbency drop in each group, but fault positive and fault negative did not appear. Method 2,absorbency and positive ratio showed no significantly difference compared with control group. Conclusion Operation time difference can cause fault positive influence of anti-HBc detection and can avoid it by pulsing enzyme reagent in turn.