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使用简并逆转录酶PCR引物对埃可病毒30分离株进行型特异性检测。

Type-specific detection of echovirus 30 isolates using degenerate reverse transcriptase PCR primers.

作者信息

Kilpatrick D R, Quay J, Pallansch M A, Oberste M S

机构信息

Respiratory and Enteric Viruses Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.

出版信息

J Clin Microbiol. 2001 Apr;39(4):1299-302. doi: 10.1128/JCM.39.4.1299-1302.2001.

Abstract

Following an approach used to specifically identify polioviruses and enterovirus 71, we have developed reverse transcriptase (RT) PCR primers containing mixed-base residues or deoxyinosine at positions of codon degeneracy. These primers permit specific RT-PCR amplification of echovirus 30 (E30) sequences by targeting sites that encode conserved amino acid motifs within the major capsid protein, VP1. All 221 E30 strains tested, isolated in 16 countries over a 44-year period, yielded the predicted 158-bp PCR product. No specific products were obtained by PCR assays containing templates from any of the other 63 EV serotypes. Inosine-containing degenerate primers may be widely applicable to the identification of echovirus serotypes by PCR.

摘要

采用一种专门用于鉴定脊髓灰质炎病毒和肠道病毒71型的方法,我们开发了在密码子简并位点含有混合碱基残基或脱氧肌苷的逆转录酶(RT)PCR引物。这些引物通过靶向编码主要衣壳蛋白VP1内保守氨基酸基序的位点,允许对埃可病毒30型(E30)序列进行特异性RT-PCR扩增。在44年期间从16个国家分离出的所有221株E30毒株,均产生了预测的158 bp PCR产物。包含来自其他63种肠道病毒血清型中任何一种模板的PCR检测均未获得特异性产物。含肌苷的简并引物可能广泛适用于通过PCR鉴定埃可病毒血清型。

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