孕妇及新生儿巨细胞病毒感染的聚合酶链反应及限制酶分析

Polymerase chain reaction and restriction enzyme analysis of human cytomegalovirus infection in pregnant women and neonates

目的 建立套式 聚合酶链反应 (Nested 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Nested PCR检测HCMVDNA ,同时结合病毒分离检测临床标本。结果 2 3例新生儿肝炎综合征患儿中 ,10例病毒分离及Nested PCR均阳性 ;1例病毒分离阴性 ,但Nested PCR阳性。对 5 8例妊娠早期孕妇血标本进行检测 ,Nested PCR阳性率为 9% ,病毒分离阳性率则为 7%。结论 Nested

Objective To establish an analytic method which combines nested PCR and restriction enzyme analysis to detect human cytomegalovirus (HCMV). Methods We designed two primers according to the sequence of HCMV immediate early protein EcoRIJ gene to detect HCMV DNA using nested PCR. Meanwhile, virus isolations were also conducted to test clinical samples. Results Among 23 cases of neonatal hepatitis syndrome, HCMV was found in 10 cases using nested PCR and virus isolation. One case showed only positive with nested PCR. Among 58 cases of early gravida, the positive rates of nested PCR and virus isolation were 8.6% and 6.9% respectively. Conclusions Nested PCR combined with restriction enzyme analysis would be effective in rapid detection of HCMV.