PCR技术检测尿中人巨细胞病毒的方法学研究

廖利民; 石炳毅; 梁春泉; 庄玉辉; 沈荣森

Abstract

廖利民,石炳毅,梁春泉,庄玉辉,沈荣森.PCR技术检测尿中人巨细胞病毒的方法学研究[J].Chinese journal of Epidemiology,1995,16(2):111-113

PCR技术检测尿中人巨细胞病毒的方法学研究

Detection of Human Cytomegalovirus in Urine by Polymerase Chain Reaction

Received:May 16, 1994 Revised:August 16, 1994

DOI：

KeyWord: 巨细胞病毒肾移植聚合酶链反应

English Key Word: HCMV Renal transplantation PCR

FundProject:

Author Name	Affiliation
Liao Li-min	Dept. of Urology, The Beijing 309 th Hospital, Beijing 100091
Shi Bing-yi	Dept. of Urology, The Beijing 309 th Hospital, Beijing 100091
Liang Chun-quan	Dept. of Urology, The Beijing 309 th Hospital, Beijing 100091
庄玉辉	中国人民解放军结核病研究中心
沈荣森	中国人民解放军军事医学科学院二所PCR实验室

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Abstract:

应用HCMV早、晚期两对DNA引物,建立了聚合酶链反应(PCR)检测尿中人巨细胞病毒(HCMV)的方法。肾移植术后HCMV感染的临床尿样检测结果表明,该方法具有快速、特异性强、灵敏度高等优点,与血清HCMV-IgM检测结果符合率为83.3%。使用早、晚期基因组的双引物分别对临床尿样进行PCR扩增,减少了由于自然点突变所致的假阴性,且引物与HCMV同属其它病毒及组织DNA间无交叉反应;以HCMV组织培养液作模板,灵敏度可达625fg。本方法为诊断肾移植受者HCMV感染提供了新的手段,适于临床推广。

English Abstract:

Polymerase chain reaction (PCR) amplification was set up with double primer pairs of major immediateearly and late gene in order to detect human cytomegalovirus (HCMV) from urine. This method was applied to the detection of HCMV in clinical samples of urine from renal transplant recipients. The result indicated that the primers did not cross react with other members of the herpes family of virus and human genomic DNA; that the HCMV AD169 tissue culture mixture used and the dilutions to estimate the sensitivity of PCR relative to tissue culture, 2.5μl of a 10-3 dilution of the culture (625fg HCMV DNA) assayed were detected by direct gel analysis; that 25 of 30 samples of urine from the renal transplant recipients (serum HCMV IgM positive) were positive. It is conclude that PCR amplification is a valuable tool for diagnoses of HCMV infection in renal transplant recipients.

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