聚合酶链反应限制酶切分析及病毒分离检测巨细胞病毒感染

The Detection of Human Cytomegalovirus in Gravida with the Early Trimester in Pregnancy and Neonates by Nested Polymerase Chain Reaction with Restriction Endonuclease and Virus Isolation

在人类巨细胞病毒（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。结果发现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９ ％ ，病毒分离阳性率则为７ ％ 。结论显示套式ＰＣＲ 加限制性内切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。

To establish a new rapid method for combination of nested polymerase chain reaction(PCR) with restriction endonuclease digestion to detect the human cytomegalovirus (HCMV),we designed two primer sequences from the DNA fragment which coding the immediate early protein Eco RIJ,and established nested PCR for detecting HCMV DNA.Meanwhile,the nested PCR and virus isolation were used to detect the HCMV in clinical samples.The results showed that 10 out of 23 sera samples from neonates with neonatal hepatitis syndrome were positive HCMV by using nested PCR and virus isolation method,one of them was negative with virus isolation,but it was positive with nested PCR technique.Among 58 samples of early gravida,the HCMV positive retes of nested PCR and virus isolation were 8.6% and 6.9%;respectively.We believe that the nested PCR combined with restriction endonuclease digestion could be useful for rapid detection of the HCMC in clinical samples test.