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利用聚合酶链反应DNA扩增技术检测新生儿尿液中的巨细胞病毒。

Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification.

作者信息

Demmler G J, Buffone G J, Schimbor C M, May R A

机构信息

Department of Pediatrics, Baylor College of Medicine, Texas Children's Hospital, Houston 77030.

出版信息

J Infect Dis. 1988 Dec;158(6):1177-84. doi: 10.1093/infdis/158.6.1177.

Abstract

Polymerase chain reaction (PCR) amplification was used to detect cytomegalovirus (CMV) in tissue culture and in urine specimens from newborns. Synthetic oligonucleotide primer pairs were used to amplify DNA from the major immediate-early and the late antigen genes of CMV. Amplified products were detected by gel electrophoresis and by dot-blot hybridization with oligonucleotide probes. Using one or both of the primer pairs and associated probes, we found 46 different tissue culture isolates of CMV that were positive; no reaction products were detected when the same primers and probes were used to amplify other herpes family viruses or human genomic DNA. Urine samples from 44 congenitally infected infants were positive when tested with one or both primer pairs and probes. When compared with tissue culture, detection by gel electrophoresis provided a sensitivity of 93%, a specificity of 100%, and a predictive value of a positive result of 100%. Dot-blot analysis raised the sensitivity to 100%. We conclude that PCR amplification may be a valuable tool for diagnosing congenital CMV infection.

摘要

采用聚合酶链反应(PCR)扩增技术检测组织培养物及新生儿尿液标本中的巨细胞病毒(CMV)。合成寡核苷酸引物对用于扩增CMV主要即刻早期抗原基因和晚期抗原基因的DNA。扩增产物通过凝胶电泳及与寡核苷酸探针的斑点杂交进行检测。使用一对或两对引物及相关探针,我们发现46种不同的CMV组织培养分离株呈阳性;当使用相同引物和探针扩增其他疱疹病毒家族病毒或人类基因组DNA时,未检测到反应产物。44例先天性感染婴儿的尿液样本在使用一对或两对引物及探针检测时呈阳性。与组织培养相比,凝胶电泳检测的敏感性为93%,特异性为100%,阳性结果预测值为100%。斑点杂交分析可将敏感性提高至100%。我们得出结论,PCR扩增可能是诊断先天性CMV感染的一项有价值的工具。

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