Evaluation of a DNA Microarray, the Check-Points ESBL/KPC Array, for Rapid Detection of TEM, SHV, and CTX-M Extended-Spectrum β-Lactamases and KPC Carbapenemases

被引:71
作者
Naas, T. [1 ]
Cuzon, G.
Truong, H.
Bernabeu, S.
Nordmann, P.
机构
[1] Hop Bicetre, INSERM, Serv Bacteriol Virol, Emerging Resistance Antibiot U914, F-94275 Le Kremlin Bicetre, France
关键词
REAL-TIME PCR; ROUTINE IDENTIFICATION; KLEBSIELLA-PNEUMONIAE; GENES; ENTEROBACTERIACEAE; POLYMORPHISM; ASSAY; EPIDEMIOLOGY; PERFORMANCE; SALMONELLA;
D O I
10.1128/AAC.01298-09
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Extended-spectrum beta-lactamases (ESBLs) and Klebsiella pneumoniae carbapenemases (KPC carbepenemases) have rapidly emerged worldwide and require rapid identification. The Check-Points ESBL/KPC array, a new commercial system based on genetic profiling for the direct identification of ESBL producers (SHV, TEM, and CTX-M) and of KPC producers, was evaluated. Well-characterized Gram-negative rods ( Enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter baumannii) expressing various beta-lactamases (KPC-2, SHV, TEM, and CTX-M types) were used as well as wild-type reference strains and isolates harboring beta-lactamase genes not detected by the assay. In addition, phenotypically confirmed ESBL producers isolated in clinical samples over a 3-month period at the Bicetre hospital were analyzed using the Check-Points ESBL/KPC array and by standard PCR. The Check-Points ESBL/KPC array allowed fast detection of all TEM, SHV, and CTX-M ESBL genes and of the KPC-2 gene. The assay allowed easy differentiation between non-ESBL TEM and SHV and their ESBL derivatives. None of the other tested beta-lactamase genes were detected, underlining its high specificity. The technique is suited for Enterobacteriaceae but also for P. aeruginosa and A. baumannii. However, for nonfermenters, especially P. aeruginosa, a 1: 10 dilution of the total DNA was necessary to detect KPC-2 and SHV-2a genes reliably. The Check-Points ESBL/KPC array is a powerful high-throughput tool for rapid identification of ESBLs and KPC producers in cultures. It provided definitive results within the same working day, allowing rapid implementation of isolation measures and appropriate antibiotic treatment. It showed an interesting potential for routine laboratory testing.
引用
收藏
页码:3086 / 3092
页数:7
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