乙型肝炎两对半酶联免疫吸附试验检测条件的优化及实验研究

Optimizing of two semi hepatitis B enzyme linked immunosorbent assay

目的分析和探讨本院乙型肝炎两对半酶联免疫吸附试验(ELISA)检测条件的优化途径。方法比较洗板机洗板后用蒸馏水或干净自来水冲洗反应板与不冲洗反应板的临界值率。结果 1洗板后用蒸馏水再次冲洗反应板数秒后、用干净自来水再次冲洗反应板与不冲洗反应板的表面抗原临界值率(临界阴性和弱阳性)分别为0.52%、2.60%和5.56%;表面抗体临界值率(临界阴性和弱阳性)分别为5.52%、8.02%和14.86%;e抗原临界值率(临界阴性和弱阳性)分别为2.82%、2.92%和3.55%。2表面抗原三者两两比较差异有统计学意义(P<0.05),表面抗体三者两两比较差异有统计学意义(P<0.05),e抗原三者两两比较差异无统计学意义(P>0.05)。结论 1洗板结束后用蒸馏水或干净自来水再次冲洗反应孔边缘可降低因酶标试剂或显色剂污染反应孔边缘导致的高临界值率结果。2用蒸馏水冲洗比用干净自来水更适合作为反应孔边缘的冲洗液。

Objective To analyze and approach the optimizing of two semi hepatitis B enzyme linked immuno-sorbent assay （ELISA）. Methods To compare the critical value rate of flushing with distilled water, flushing with cleaning tap water and flushing without anything. Results The HBsAg critical value rate of flushing with distilled water, flushing with cleaning tap water and flushing without anything was 0.52%, 2.60% and 5.56%; The HBsAb critical value rate of flushing with distilled water, flushing with cleaning tap water and flushing without anything was 5.52%, 8.02%and 14.86%; The HBeAg critical value rate of flushing with distilled water, flushing with cleaning tap water and flushing without anything was 2.82%, 2.92%and 3.55%. The HBsAg critical value rate had distinct differentiation （P〈0.05）;The HBsAb critical value rate had distinct differentiation（P〈0.05）; The consistency of HBeAg was well（P〈0.05）. Conclusion Flushing with distilled water or flushing with cleaning tap water can reduce the high critical value rate because of enzyme or developer pollution. Flushing with distilled water exhibits more efficacy than flushing with cleaning tap water.