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[应用聚合酶链反应体外基因扩增技术快速检测尿样中人类巨细胞病毒DNA]

[Rapid detection of human cytomegalovirus DNA in urine samples by polymerase chain reaction in vitro gene amplification].

作者信息

Dong F, Chen S

出版信息

Zhonghua Yi Xue Za Zhi. 1990 Oct;70(10):560-2, 40.

PMID:2176565
Abstract

In the detection of human cytomegalovirus (HCMV) DNA in the urine by polymerase chain reaction (PCR). We have designed and synthesized two pairs of oligonucleotide primers (patent pending) corresponding to the specific and conserved DNA sequences of HCMV AD 169 strain. The sequences of the primers were checked that they did not have cross reaction with either normal human genomic DNA or with other herpesviruses. As less as 1fg of HCMV DNA (equal to the amount of DNA from four HCMV virions) in urine can be detected by the method A total of 47 urine specimens from infants suspected of HCMV infection was examined by PCR and conventional virus isolation simultaneously 31 samples were positive in both PCR and virus isolation; 12 samples were negative in either; 4 samples were positive in PCR but negative in virus isolation. So the samples were further examined by serology and the data showed that there was HCMV-specific IgM in all the virus isolation negative sera examined. The result suggests that HCMV in the above samples is possibly underdetected by virus isolation. Therefore the method described in the paper is a specific, sensitive, rapid conventional and non radioisotopic one for detection of HCMV in urine samples.

摘要

在通过聚合酶链反应(PCR)检测尿液中的人巨细胞病毒(HCMV)DNA时。我们设计并合成了两对与HCMV AD 169株的特异性和保守DNA序列相对应的寡核苷酸引物(专利申请中)。对引物序列进行了检查,确保它们与正常人基因组DNA或其他疱疹病毒均无交叉反应。该方法可检测到尿液中低至1fg的HCMV DNA(相当于四个HCMV病毒粒子的DNA量)。同时对47份疑似HCMV感染婴儿的尿液标本进行了PCR检测和传统病毒分离。31份样本PCR和病毒分离均呈阳性;12份样本两者均为阴性;4份样本PCR呈阳性但病毒分离呈阴性。因此对这些样本进一步进行血清学检查,数据显示在所有检测的病毒分离阴性血清中均存在HCMV特异性IgM。结果表明上述样本中的HCMV可能未被病毒分离法充分检测到。因此本文所述方法是一种用于检测尿液样本中HCMV的特异性、灵敏、快速、传统且非放射性同位素的方法。

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