套式聚合酶链反应及限制酶分析检测新生儿肝炎巨细胞病毒感染

在人巨细胞病毒（HCMV）AD169株直接早期蛋白EcoRIJDNA片段内，自行设计二对引物，建立套式PCR检测HCMVDNA。经实验证明有高度的特异性与敏感性，对其它疙疹类病毒和正常人基因组DNA无交叉反应。一次性PCR检测HCMVAD169株基因组的最小检出量为100fg，而套式PCR可达10fg，经PStI酶切后，得到的片段与设计相符。对23例新生儿肝炎临床标本进行病毒分离，一次性PCR，套式PCR检测，结果表明套式PCR能进一步提高一次性PCR的特异性与敏感性，适用于新生儿肝炎HCMV感染的临测。

STUDY OF CYTOMEGAIOVIRAL INFECTION IN NEONATAL HEPATITIS WITH NESTED POLYMERASE CHAIN REACTION (nPCE)

nPCR was developed to detect conserved sequence from the immediate early gene of human cytomegalovirus (HCMV). The two primer sequences were from EcoRI J fragment of AD 169. The experiments showed that nPCR has highly speciflcity and sensitivity. There was no cross-reaction with human DNA or with DNA from other herpes viruses. With single PCR, it was possible to detect 100fg HCMV DNA, but with nPCR. 10fg were detectable, which further used by PstI endonucleases analyses was in accordance with the designed one. 23 clinic samples were analysed by single PCR. nPCR and virus isolation. The results suggest that nPCR may he used for rapid detection of HCMV DNA in the neonates with hepatitis of HCMV infection.