人巨细胞病毒基因工程糖蛋白52kd抗原的临床初步应用

作者在构建人巨细胞病毒(HCMV)糖蛋白52 kd抗原编码区段的表达质粒pHCMV-52基础上,从含该克隆基因的大肠杆菌中分离并纯化其表达产物作为抗原,与已知ELISA-HCMV-IgM为阳性(12例)和阴性(4例)的血清反应,采用辣根过氧化物酶(HRP)标记的马抗人抗体取代同位素检测特异抗原—抗体复合物,两种方法结果一致者10例(10/12)。该临床初步结果显示,基因工程技术制备HCMV抗原可用于HCMV感染的临床诊断,并具有抗原制备方便、产量高、临床实用和易于推广等优点。

Preliminary Use of Recombinant Glycoprotein 52kd as an Antigen in the Diagnosis of Human Cytomegalovirus Infection

Interest in the human cyto-megalovirus(HCMV) mainly derives fromits associations with congenital malforma-tions, mental retardation, and severe orfatal infections in immunosuppressed indi-viduals such as transplant patients, tumorand AIDS patients. It is evidenced thatthere has been a need for a rapid and sen-sitive methods to detect an ongoing acuteinfection. The recent studies showed thatnigh titers of antibody to the glycoprotein52kd are present in sera of patients under-going acute HCMV infection. However,purification of individual glycoprotein fromHCMV-infected cells is a daunting prospect.HCMV glycoprotein 52 kd expressed via re-combinant DNA techniques are a promisingapproach to solve this problem. In orderto evaluate the diagnostic value of the re-combinant glycoprotein 52 kd antigenic coderegion for HCMV infection, we have usedthe polymerase chain reaction(PCR) andrecombinant DNA techniques to constructsuccessfully the high-level expression plas-mid pHCMV containing the HCMV GP-52kd antigenic code region,with the predict-ed protein at levels up to 20% in total bac-terial protein. The expressed protein waspurified from SDS-PAGE, used as an anti-gen in Western-blot, and reacted with 12cases of the positive sera, 4 cases of thenegative sera, following by reaction withHRP-labelled horse IgG antibody againsthuman. The results indicated that the ap-proach we are using to detect antibody toHCMV acute infection are as sensitive asgeneral serological mathods such as ELISA,with the advantages of easy preparation ofantigen with high quantity, and clinicalpracticability.