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	Multiplex asymmetric PCR-based oligonucleotide microarray for detection of drug resistance genes containing single mutations in Enterobacteriaceae
	Zhu, Ling-Xiang ; Zhang, Zhi-Wei ; Liang, Dong ; Jiang, Di ; Wang, Can ; Du, Ning ; Zhang, Qiong ; Mitchelson, Keith ; Cheng, Jing
	2010-05-11 ; 2010-05-11 ; OCT
关键词	SPECTRUM BETA-LACTAMASES THE-EXPONENTIAL (LATE)-PCR EXTENDED-SPECTRUM KLEBSIELLA-PNEUMONIAE ANTIBIOTIC-RESISTANCE DNA EPIDEMIOLOGY IDENTIFICATION COMBINATION INFECTIONS Microbiology Pharmacology & Pharmacy
中文摘要	A multiplex asymmetric PCR (MAPCR)-based microarray method was developed for the detection of 10 known extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC P-lactamase genes in gram-negative bacteria and for the typing of six important point mutations (amino acid positions 35, 43, 130, 179, 238, and 240) in the blas(SHV) gene. The MAPCR is based on a two-round reaction to promote the accumulation of the single-stranded amplicons amenable for microarray hybridization by employing multiple universal unrelated sequence-tagged primers and elevating the annealing temperature at the second round of amplification. A strategy to improve the discrimination efficiency of the microarray was constituted by introducing an artificial mismatch into some of the allele-specific oligonucleotide probes. The microarray assay correctly identified the resistance genes in both the reference strains and some 111 clinical isolates, and these results were also confirmed for some isolates by direct DNA sequence analysis. The resistance genotypes determined by the microarray correlated closely with phenotypic MIC susceptibility testing. This fast MAPCR-based microarray method should prove useful for undertaking important epidemiological studies concerning ESBLs and plasmid-mediated AmpC enzymes and could also prove invaluable as a preliminary screen to supplement phenotypic testing for clinical diagnostics.
语种	英语 ; 英语
出版者	AMER SOC MICROBIOLOGY ; WASHINGTON ; 1752 N ST NW, WASHINGTON, DC 20036-2904 USA
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/27565]
专题	清华大学
推荐引用方式 GB/T 7714	Zhu, Ling-Xiang,Zhang, Zhi-Wei,Liang, Dong,et al. Multiplex asymmetric PCR-based oligonucleotide microarray for detection of drug resistance genes containing single mutations in Enterobacteriaceae[J],2010, 2010, OCT.
APA	Zhu, Ling-Xiang.,Zhang, Zhi-Wei.,Liang, Dong.,Jiang, Di.,Wang, Can.,...&Cheng, Jing.(2010).Multiplex asymmetric PCR-based oligonucleotide microarray for detection of drug resistance genes containing single mutations in Enterobacteriaceae..
MLA	Zhu, Ling-Xiang,et al."Multiplex asymmetric PCR-based oligonucleotide microarray for detection of drug resistance genes containing single mutations in Enterobacteriaceae".(2010).

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