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用于检测肠道病毒的通用引物介导聚合酶链反应:在诊断常规感染和持续性感染中的应用

General primer-mediated polymerase chain reaction for detection of enteroviruses: application for diagnostic routine and persistent infections.

作者信息

Zoll G J, Melchers W J, Kopecka H, Jambroes G, van der Poel H J, Galama J M

机构信息

Department of Medical Microbiology, University of Nijmegen, The Netherlands.

出版信息

J Clin Microbiol. 1992 Jan;30(1):160-5. doi: 10.1128/jcm.30.1.160-165.1992.

Abstract

The aim of this study was to determine the applicability of the polymerase chain reaction (PCR) for routine diagnostic use and for the detection of persistent enteroviral infections. To this end, general primers were selected in the highly conserved part of the 5'-noncoding region of the enteroviral genome. They were tested on 66 different enterovirus serotypes. A specific fragment was amplified from 60 of 66 serotypes. An amplification product was not observed from coxsackievirus types A11, A17, and A24 and echovirus types 16, 22, and 23. Enteroviral RNA was detected by the PCR in routinely collected throat swabs and stool specimens that were found to be positive for enterovirus by isolation in tissue culture. Enteroviral RNA was detected in one of five myocardial biopsy specimens from patients with dilated cardiomyopathy, implicating virus persistence. No amplification product was obtained from eight control samples. Our results demonstrate the significance of the PCR for the detection of enteroviral RNA and, in particular, for the demonstration of persistent enteroviral infections.

摘要

本研究的目的是确定聚合酶链反应(PCR)在常规诊断用途以及检测持续性肠道病毒感染方面的适用性。为此,在肠道病毒基因组5'-非编码区的高度保守部分选择了通用引物。它们在66种不同的肠道病毒血清型上进行了测试。从66种血清型中的60种扩增出了特定片段。未从柯萨奇病毒A11型、A17型和A24型以及埃可病毒16型、22型和23型中观察到扩增产物。通过PCR在常规收集的咽拭子和粪便标本中检测到肠道病毒RNA,这些标本经组织培养分离发现对肠道病毒呈阳性。在扩张型心肌病患者的五份心肌活检标本中的一份中检测到肠道病毒RNA,提示病毒持续存在。从八个对照样本中未获得扩增产物。我们的结果证明了PCR在检测肠道病毒RNA方面的重要性,特别是在证明持续性肠道病毒感染方面。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3438/265013/1a0e4c91fbd1/jcm00025-0184-a.jpg

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