Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA-1 genes in Enterobacteriaceae
Simple and reliable multiplex PCR assay for detection of blaTEM, blaSHV and blaOXA-1 genes in Enterobacteriaceae
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DOI：
10.1016/s0378-1097(03)00306-9
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发表时间：
2003-06-27
影响因子：
2.1
通讯作者：
Cisterna, R
中科院分区：
文献类型：
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作者：
Colom, K;Pérez, J;Cisterna, R
肠杆菌科细菌对第三代头孢菌素耐药通常由TEM型和SHV型β-内酰胺酶介导。TEM型和OXA-1酶是引起大肠埃希菌对阿莫西林-克拉维酸耐药的主要质粒型β-内酰胺酶。我们建立了一种快速、简便的多重聚合酶链式反应(PCR)方法，通过分别产生516、392和619个片段来区分blA(TEM)、blA(SHV)和blA(OXA-1)基因。对51株阿莫西林-克拉维酸耐药的大肠埃希菌进行多重聚合酶链式反应分析，分别在45株和2株中检测到blA(TEM)和blA(SHV)基因，仅1株携带blA(OXA-1)基因。在40株对头孢噻肟耐药的肠杆菌科分离株中，有23株扩增产物的大小与bla(TEM)基因(13株)、bla(Shv)基因(6株)或两种基因的关联(4株)兼容。通过菌落杂交验证了上述结果。因此，多重聚合酶链式反应是一种适合于肠杆菌科细菌bla基因初步快速筛选的工具。(C)2003年欧洲微生物学会联合会。爱思唯尔科学公司出版。版权所有。
Third-generation cephalosporin resistance is often mediated by TEM- and SHV-type beta-lactamases in Enterobacteriaceae. TEM-type and OXA-1 enzymes are the major plasmid-borne beta-lactamases implicated in amoxicillin-clavulanic acid resistance in Escherichia coli isolates. We have developed a rapid and simple multiplex polymerase chain reaction (PCR) which discriminates bla(TEM), bla(SHV) and bla(OXA-1) genes by generating fragments of 516, 392 and 619 by respectively. Multiplex PCR analysis of 51 amoxicillin-clavulanate resistant E. coli isolates detected bla(TEM) and bla(SHV) genes in 45 and two strains, respectively, and only one strain harboured a bla(OXA-1) gene. Twenty-three of the 40 cefotaxime-resistant Enterobacteriaceae isolates produced amplicons with a size compatible with the presence of bla(TEM) (13 strains), bla(SHV) (six strains) genes or the association of both genes (four strains). These results were verified by colony hybridisation. Therefore, multiplex PCR is a suitable tool for initial rapid screening of bla genes in Enterobacteriaceae. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.