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应用多重 PCR 和高分辨率熔解曲线分析鉴定与侵袭性感染和抗生素耐药性相关的常见 A 组链球菌 emm 型。

Identification of group A streptococcal emm types commonly associated with invasive infections and antimicrobial resistance by the use of multiplex PCR and high-resolution melting analysis.

机构信息

Univ Paris Diderot, Sorbonne Paris Cité, EA 3105, Paris, France.

出版信息

Eur J Clin Microbiol Infect Dis. 2012 Oct;31(10):2817-26. doi: 10.1007/s10096-012-1635-5. Epub 2012 May 20.

Abstract

M/emm typing, based either on serotyping of the M protein or on sequencing of the emm gene, is a major tool for epidemiological studies of group A streptococci (GAS). In order to simplify M/emm typing, we designed two multiplex polymerase chain reaction (PCR) formats capable of identifying the most frequent GAS M/emm types involved in invasive infections and antimicrobial resistance. A heptaplex PCR procedure was first developed in a conventional format coupled with gel electrophoresis to identify emm types 1, 3, 4, 6, 12, 28, and 89, based on the size of the amplification products. The other method, designed to identify the same seven emm types, together with emm11, was based on a real-time PCR format coupled with high-resolution melting (HRM) analysis, allowing the rapid typing of large strain collections.

摘要

M/emm 分型,无论是基于 M 蛋白的血清型分析还是 emm 基因的测序,都是 A 组链球菌(GAS)流行病学研究的主要工具。为了简化 M/emm 分型,我们设计了两种多重聚合酶链反应(PCR)格式,能够鉴定与侵袭性感染和抗生素耐药性相关的最常见的 GAS M/emm 型。首先,我们开发了一种七重 PCR 程序,采用常规格式结合凝胶电泳,根据扩增产物的大小鉴定 emm 型 1、3、4、6、12、28 和 89。另一种方法旨在鉴定相同的七种 emm 型,以及 emm11,基于实时 PCR 格式结合高分辨率熔解(HRM)分析,允许快速对大量菌株进行分型。

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