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目的 :建立用于诊断HCMV活动性感染的原位免疫PCR方法。方法 :HCMVAD16 9株感染的人胚肺成纤维细胞或先天畸形胎儿肾组织石蜡切片 ,依次加入抗HCMV单抗、生物素化二抗、链霉亲合素以及作为指示元件的生物素化DNA ,随后进行原位PCR扩增 ,经酶显色反应判断结果。结果 :应用本法检测HCMV感染的细胞可于感染后 8h清晰检出阳性信号 ;检测先天畸形胎儿肾组织HCMV感染 ,阳性率显著高于免疫组化法。结论 :本方法具有敏感性高、特异性好及精确组织定位等特点 ,可用于HCMV活动性感染诊断
Objective: To establish an in situ immuno-PCR method for diagnosis of active HCMV infection. Methods: HCMVAD16 infected human embryonic lung fibroblasts or congenital malformation fetus kidney tissue paraffin section, followed by anti-HCMV monoclonal antibody, biotinylated secondary antibody, streptavidin and as an indicator element biotinylated DNA, Followed by in situ PCR amplification, the enzyme chromogenic reaction to determine the results. Results: HCMV infected cells were detected by this method 8 h after infection clear positive signals detected; detection of congenital malformation fetal kidney HCMV infection, the positive rate was significantly higher than immunohistochemistry. Conclusion: The method has the characteristics of high sensitivity, good specificity and precise tissue localization, which can be used in diagnosis of active HCMV infection