Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae

被引:1363
作者
Dallenne, Caroline [1 ]
Da Costa, Anaelle [1 ]
Decre, Dominique [1 ,2 ]
Favier, Christine [3 ]
Arlet, Guillaume [1 ,4 ]
机构
[1] Univ Paris 06, Fac Med Pierre & Marie Curie, Dept Bacteriol, Paris, France
[2] Hop St Antoine, Hop Paris, Microbiol Serv, F-75571 Paris, France
[3] Biorad Labs Inc, Marnes La Coquette, France
[4] Hop Tenon, Hop Paris, Serv Bacteriol, F-75970 Paris, France
关键词
OXA-1-like beta-lactamases; ESBLs; plasmid-mediated AmpC beta-lactamases; carbapenemases; molecular epidemiology; RAPID DETECTION;
D O I
10.1093/jac/dkp498
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
To develop a rapid and reliable tool to detect by multiplex PCR assays the most frequently widespread beta-lactamase genes encoding the OXA-1-like broad-spectrum beta-lactamases, extended-spectrum beta-lactamases (ESBLs), plasmid-mediated AmpC beta-lactamases and class A, B and D carbapenemases. Following the design of a specific group of primers and optimization using control strains, a set of six multiplex PCRs and one simplex PCR was created. An evaluation of the set was performed using a collection of 31 Enterobacteriaceae strains isolated from clinical specimens showing a resistance phenotype towards broad-spectrum cephalosporins and/or cephamycins and/or carbapenems. Direct sequencing from PCR products was subsequently carried out to identify beta-lactamase genes. Under optimized conditions, all positive controls confirmed the specificity of group-specific PCR primers. Except for the detection of carbapenemase genes, multiplex and simplex PCR assays were carried out using the same PCR conditions, allowing assays to be performed in a single run. Out of 31 isolates selected, 22 strains produced an ESBL, mostly CTX-M-15 but also CTX-M-1 and CTX-M-9, SHV-12, SHV-5, SHV-2, TEM-21, TEM-52 and a VEB-type ESBL, 6 strains produced a plasmid-mediated AmpC beta-lactamase (five DHA-1 and one CMY-2) and 3 strains produced both an ESBL (two SHV-12, one CTX-M-15) and a plasmid-mediated AmpC beta-lactamase (DHA-1). We report here the development of a useful method composed of a set of six multiplex PCRs and one simplex PCR for the rapid screening of the most frequently encountered beta-lactamases. This method allowed direct sequencing from the PCR products.
引用
收藏
页码:490 / 495
页数:6
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