用于检测肠杆菌科细菌中blaSHV、blaTEM和blaCTX-M基因的多重PCR扩增检测法。

Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae.

作者信息

Monstein H-J, Ostholm-Balkhed A, Nilsson M V, Nilsson M, Dornbusch K, Nilsson L E

机构信息

Division of Clinical Microbiology, University Hospital Linköping, Linköping, Sweden.

出版信息

APMIS. 2007 Dec;115(12):1400-8. doi: 10.1111/j.1600-0463.2007.00722.x.

DOI:10.1111/j.1600-0463.2007.00722.x

PMID:18184411

Abstract

Extended-spectrum beta-lactamases (ESBLs) are often mediated by (bla-)SHV, (bla)TEM and (bla)CTX-M genes in Enterobacteriaceae and other Gram-negative bacteria. Numerous molecular typing methods, including PCR-based assays, have been developed for their identification. To reduce the number of PCR amplifications needed we have developed a multiplex PCR assay which detects and discriminates between (bla-)SHV, (bla)TEM and (bla)CTX-M PCR amplicons of 747, 445 and 593 bp, respectively. This multiplex PCR assay allowed the identification of (bla-)SHV, (bla)TEM and (bla)CTX-M genes in a series of clinical isolates of Enterobacteriaceae with previously characterised ESBL phenotype. The presence of (bla)SHV, (bla)TEM and (bla)CTX-M genes was confirmed by partial DNA sequence analysis. Apparently, the universal well-established CTX-M primer pair used here to reveal plasmid-encoded (bla)CTX-M genes would also amplify the chromosomally located K-1 enzyme gene in all Klebsiella oxytoca strains included in the study.

摘要

超广谱β-内酰胺酶（ESBLs）通常由肠杆菌科细菌及其他革兰氏阴性菌中的（bla-）SHV、（bla）TEM和（bla）CTX-M基因介导。包括基于PCR的检测方法在内，已经开发出多种分子分型方法用于其鉴定。为减少所需的PCR扩增次数，我们开发了一种多重PCR检测方法，该方法可分别检测和区分大小为747 bp、445 bp和593 bp的（bla-）SHV、（bla）TEM和（bla）CTX-M PCR扩增子。这种多重PCR检测方法能够在一系列具有先前已鉴定的ESBL表型的肠杆菌科临床分离株中鉴定出（bla-）SHV、（bla）TEM和（bla）CTX-M基因。通过部分DNA序列分析证实了（bla）SHV、（bla）TEM和（bla）CTX-M基因的存在。显然，此处用于揭示质粒编码的（bla）CTX-M基因的通用且成熟的CTX-M引物对，也会扩增本研究中所有产酸克雷伯菌菌株中位于染色体上的K-1酶基因。