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目的建立套式聚合酶链反应(PCR)加限制性酶切分析方法检测人巨细胞病毒(HCMV)。方法在HCMV直接早期蛋白EcoRIJDNA片段内,自行设计两对引物,建立套式PCR检测HCMVDNA,同时结合病毒分离检测临床标本。结果23例新生儿肝炎综合征患儿中,10例病毒分离及套式PCR均阳性;1例病毒分离阴性,但套式PCR阳性。对58例妊娠早期孕妇血标本进行检测,套式PCR阳性率为9%,病毒分离阳性率则为7%。结论套式PCR加限制性酶切分析是一种临床检测HCMV的快速有效手段,值得临床推广。
Objective To establish a nested polymerase chain reaction (PCR) and restriction enzyme digestion assay to detect human cytomegalovirus (HCMV). Methods Two pairs of primers were designed in HCMV direct early EcoRIJDNA fragment to establish nested PCR for detection of HCMV DNA. Meanwhile, clinical isolates were detected by virus isolation. Results Of the 23 cases of neonatal hepatitis syndrome, 10 cases were positive for virus isolation and nested PCR. One case was negative for virus isolation but positive for nested PCR. 58 cases of pregnant women in early pregnancy blood samples were detected, nested PCR positive rate was 9%, the positive rate of virus isolation was 7%. Conclusion Nested PCR plus restriction analysis is a rapid and effective method for clinical detection of HCMV and is worthy of clinical promotion.