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目的：依据免疫－ＰＣＲ基本原理，建立检测人巨细胞病毒（ＨＣＭＶ）活动性感染的免疫－ＰＣＲ方法。方法：我们建立的免疫－ＰＣＲ方法与酶联免疫ＥＬＩＳＡ不同之处是，用ＰＣＲ扩增生物素化线性ｐＢＶ２２０ＤＮＡ代替ＥＬＩＳＡ的酶催化底物，经琼脂糖凝胶电泳溴化乙锭染色观察分析扩增产物特异条带，判断结果。结果：用本法检测ＨＣＭＶ的敏感性高于ＥＬＩＳＡ法１０３～１０５倍，可检测到５个感染ＨＣＭＶ细胞；而检测ＨＳＶ－１、ＶＺＶ、ＢＫＶ均呈阴性反应，显示良好特异性。结论：建立的免疫－ＰＣＲ方法具有高度敏感性，可用于检测微量ＨＣＭＶ抗原，诊断ＨＣＭＶ活动性感染
OBJECTIVE: To establish an immuno-PCR method for detecting active human cytomegalovirus (HCMV) infection based on the basic principles of immune-PCR. Methods: The immune-PCR method we established was different from enzyme-linked immunosorbent assay (ELISA) in that the enzyme-catalyzed substrate of biotinylated linearized pBV220 DNA instead of ELISA was amplified by PCR and analyzed by agarose gel electrophoresis Increased product specific bands to determine the results. Results: The sensitivity of this method to detect HCMV was 103- to 105-fold higher than that of ELISA method. Five HCMV infected cells could be detected. However, HSV-1, VZV and BKV were negative and showed good specificity. Conclusion: The established immune-PCR method is highly sensitive and can be used to detect trace HCMV antigen to diagnose active HCMV infection