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通过聚合酶链反应鉴定人巨细胞病毒分离株。

Identification of human cytomegalovirus isolates by the polymerase chain reaction.

作者信息

Zipeto D, Silini E, Parea M, Percivalle E, Zavattoni M, Di Matteo A, Milanesi G, Gerna G

机构信息

Istituto di Genetica, Biochimica ed Evoluzionistica del C.N.R., University of Pavia, Italy.

出版信息

Microbiologica. 1990 Oct;13(4):297-304.

PMID:1965006
Abstract

Fifty human cytomegalovirus (HCMV) isolates were recovered from different clinical specimens (buffy coat, throat washing and urine) obtained from fifty patients (23 AIDS patients, 20 heart transplant recipients, 1 bone marrow transplant recipient, 2 newborns with congenital HCMV infection and 4 immunocompetent individuals with acute HCMV infection). The isolates were previously identified by immunological methods and then examined for identification by the polymerase chain reaction. In parallel, reference HCMV strains as well as other human members of the Herpesviridae family (reference and wild strains) were examined as controls. Two pairs of primers relevant to the immediate-early and late regions of HCMV DNA, respectively, were used. The DNA amplification product was highly specific; in addition, all fifty HCMV isolates were amplified by both pairs of primers and thus identified as HCMV. These preliminary results show that selected pairs of primers are able to amplify DNA regions conserved enough to allow virus identification among a large number of HCMV strains. In addition, they are highly promising in view of the use of PCR for direct detection of HCMV DNA in clinical samples.

摘要

从50名患者(23名艾滋病患者、20名心脏移植受者、1名骨髓移植受者、2名先天性人巨细胞病毒感染的新生儿和4名具有急性人巨细胞病毒感染的免疫功能正常个体)的不同临床标本(血沉棕黄层、咽洗液和尿液)中分离出50株人巨细胞病毒(HCMV)毒株。这些毒株先前已通过免疫学方法鉴定,然后通过聚合酶链反应进行鉴定检查。同时,将人巨细胞病毒参考毒株以及疱疹病毒科的其他人类成员(参考毒株和野生毒株)作为对照进行检测。分别使用了与HCMV DNA即刻早期和晚期区域相关的两对引物。DNA扩增产物具有高度特异性;此外,所有50株HCMV毒株均能被这两对引物扩增,因此被鉴定为人巨细胞病毒。这些初步结果表明,所选的引物对能够扩增出足够保守的DNA区域,以便在大量HCMV毒株中进行病毒鉴定。此外,鉴于使用聚合酶链反应直接检测临床样本中的HCMV DNA,它们具有很大的前景。

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