应用增强化学发光法检测PCR扩增的人巨细胞病毒核酸

本文报道用增强化学发光法（ＥＣＬ）检测ＰＣＲ扩增的人巨细胞病毒（ＨＣＭＶ）核酸。该检测系统用戊二醛将辣根过氧化物酶复合物（ＨＲｐ－ＰＢＱ－ＰＥＩ＋－ＮＨ３＋）与探针结合制成酶标基因探针。探针与靶ＤＮＡ结合后ＨＲＰ可催化检测系统中的发光底物，经增强剂将光信号放大，杂交信号通过Ｘ光片显影。实验证明，用ＥＣＬ检测ＰＣＲ扩增的ＨＣＭＶＤＮＡ产物，敏感性可达０．０１ｆｇ的ＤＮＡ片段，且仅与ＨＣＭＶＤＮＡ的扩增产物杂交。临床应用表明，此方法具有快速、简便、敏感、安全的特点。

DETECTYION OF THE AMPLIFIED PRODUCTS OF HUMAN CYTOMEGALOVIRUS DNA BY ENHANCED CHEMICAL LUMINESCENCE

The PCR amplified products of human cytomegalovirus(HCMV)DNA was detected by enhanced chemical lu-minescence(ECL) using probes directly conjugated with the activated horseradish peroxidase complexes(HRP-PBQ-PEI -NH3 ).The HRP bound to the probe acted on substrates hydrogen peroxide and luminol in the presenceof an enhancer,which resulted in a sustained emission of light.The chemiluminescence was recorded by exposure toX-ray film. The experiments showed that the lowest detection level was 0.0lfg DNA fragment and the probe onlyhybridlzed to the amplified products of HCMV DNA. The result of clinical application showed the methods of de-tection of PCR amplified products by ECL was rapid,simple,sensitive and specific.