PCR Classification of CTX-M-Type β-Lactamase Genes Identified in Clinically Isolated Gram-Negative Bacilli in Japan

Shibata, Naohiro; Kurokawa, Hiroshi; Doi, Yohei; Yagi, Tetsuya; Yamane, Kunikazu; Wachino, Jun Ichi; Suzuki, Satowa; Kimura, Kouji; Ishikawa, Satoshi; Kato, Haru; Ozawa, Yoshiyuki; Shibayama, Keigo; Kai, Kumiko; Konda, Toshifumi; Arakawa, Yoshichika

doi:10.1128/aac.50.2.791-795.2006

Cited by 128 publications

(71 citation statements)

References 28 publications

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“…In this study, all the isolates produced ESBLs that belonged to the CTX-M-1 group; this is consistent with the findings of recent surveys, which state that the CTX-M-1 group ESBLs are the most common and are ubiquitous in Europe (12) as well as in Japan (13). However, we cannot deny the possibility that CTX-M-1 group ESBLs may be prevalent in our hospital and horizontal transmission is ongoing.…”

supporting

confidence: 81%

Proportion of Rectal Carriage of Extended-Spectrum ^|^beta;-Lactamase-Producing Enterobacteriaceae in the Inpatients of a Pediatric Tertiary Care Hospital in Japan

et al. 2012

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supporting

confidence: 81%

Proportion of Rectal Carriage of Extended-Spectrum ^|^beta;-Lactamase-Producing Enterobacteriaceae in the Inpatients of a Pediatric Tertiary Care Hospital in Japan

et al. 2012

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“…Two isolates that had originally been shown to harbor bla SHV or bla KPC presumably lost plasmids during cryopreservation, as they were PCR negative for these genes; they were included as negative controls. Discordant results were resolved with additional conventional PCR testing (25,26). The CT103 or CT103 XL was repeated at least once if falsely negative results were obtained (i.e., discordant with the known genotype of the isolate).…”

mentioning

confidence: 99%

Evaluation of the Check-Points Check MDR CT103 and CT103 XL Microarray Kits by Use of Preparatory Rapid Cell Lysis

2016

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Using a rapid bacterial lysis method, the Check MDR CT103 and CT103 XL microarrays demonstrated accuracies of 98.1% and 94.2%, respectively, for detection of known resistance genes in 108 multidrug-resistant Gram-negative bacilli. In 45 isolates, 49 previously unrecognized extended-spectrum ␤-lactamase or plasmid AmpC targets were detected and confirmed by conventional PCR. Multidrug-resistant Enterobacteriaceae (MDRE), especially those producing carbapenemases, are of concern to physicians and laboratorians worldwide. MDRE have acquired resistance through plasmid transfer, with or without chromosomal mutation (1). The ability of clinical laboratories to rapidly detect and characterize MDRE is a sentinel defense against these threatening pathogens. Current laboratory tools directed at MDRE and their associated resistances are largely focused on phenotypic methods, such as selective indicator media (e.g., commercial chromogenic media), antimicrobial susceptibility testing (AST) systems to assess MICs, and confirmatory tests for enzyme expression (e.g., modified Hodge test, Carba NP test) (2-5).A multitude of PCR assays have been described for detecting resistance genes in isolated bacteria (6-10). Sensitivity and specificity are generally excellent; however, such assays typically target a relatively small number of ␤-lactamase genes based largely on prevalence. MDRE may carry one or more plasmids and may simultaneously harbor multiple antimicrobial resistance genes and resistance mechanisms that affect the same antimicrobial agents (11). Given this, a comprehensive approach to detect resistance genes may be appropriate for MDRE. Studies employing microarray-based molecular tests that target a large number of resistance genes suggest that such platforms may serve this role (5, 12-15). Cuzon et al. evaluated the Check-Points Check MDR CT103 (Check-Points Health B.V., Wageningen, The Netherlands) microarray kit by using 187 clinical isolates and reported excellent specificity (16). This kit can detect 6 AmpC genes/gene groups (CMY-I/MOX, ACC, DHA, ACT/MIR, CMY-II, FOX), 5 carbapenemase genes/gene groups (KPC, NDM, VIM, IMP, OXA-48-like), and genes encoding CTX-M-1, -2, -8, -9, and -25 groups and can detect and distinguish wild-type bla SHV and bla TEM alleles from those with point mutations conferring extended-spectrum ␤-lactamase (ESBL) production (16).(This study was presented in part at the 54th Interscience Conference on Antimicrobial Agents and Chemotherapy, Washington, DC, 5 to 9 September 2014.)We evaluated the Check MDR CT103 kit and the expanded CT103 XL kit (Check-Points Health B.V.) following the manufacturer's suggested protocol but substituting a previously described time-and reagent-saving rapid bacterial lysis step (6) for DNA preparation. The CT103 XL kit contains additional targets beyond those included in the CT103 kit, including carbapenemases most commonly found in nonfermenting Gram-negative bacilli rather than Enterobacteriaceae, specifically GES, GIM, SPM, OXA-23-like, OXA-24/40-like, and OXA-...

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“…En la detección de genes bla CTX-M , se utilizaron los iniciadores CTX-MA y CTX-MB para el grupo 1 (17), los iniciadores TOHO1A y TOHO1B para el grupo 2 (18) y los C1 y C2 para el grupo 9 (19). Para la detección de los genes bla CTX-M del grupo 8 se utilizaron los iniciadores CTX-M-8-F y CTX-M-8-R (20). Se realizaron mezclas de reacción con concentraciones de 0,2 mM de dNTPs, 1,5 mM de MgCl 2 , 1 µM cada iniciador y 0,04 U/µl unidades de TaqDNA polimerasa (Invitrogen®).…”

Section: Detección De Genes Blaunclassified

Distribución de genes codificadores de β-lactamasas de espectro extendido en aislamientos de Klebsiella pneumoniae de hospitales de Bogotá, D.C., Colombia

et al. 2011

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PCR Classification of CTX-M-Type β-Lactamase Genes Identified in Clinically Isolated Gram-Negative Bacilli in Japan

Cited by 128 publications

References 28 publications

Proportion of Rectal Carriage of Extended-Spectrum ^|^beta;-Lactamase-Producing Enterobacteriaceae in the Inpatients of a Pediatric Tertiary Care Hospital in Japan

Proportion of Rectal Carriage of Extended-Spectrum ^|^beta;-Lactamase-Producing Enterobacteriaceae in the Inpatients of a Pediatric Tertiary Care Hospital in Japan

Evaluation of the Check-Points Check MDR CT103 and CT103 XL Microarray Kits by Use of Preparatory Rapid Cell Lysis

Distribución de genes codificadores de β-lactamasas de espectro extendido en aislamientos de Klebsiella pneumoniae de hospitales de Bogotá, D.C., Colombia

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