在一次实时聚合酶链反应中对blaVIM和blaIMP金属β-内酰胺酶基因进行特异性检测。
Specific detection of blaVIM and blaIMP metallo-beta-lactamase genes in a single real-time PCR.
作者信息
Bisiklis A, Papageorgiou F, Frantzidou F, Alexiou-Daniel S
机构信息
Department of Clinical Microbiology, AHEPA University Hospital, School of Medicine, Thessaloniki, Greece.
出版信息
Clin Microbiol Infect. 2007 Dec;13(12):1201-3. doi: 10.1111/j.1469-0691.2007.01832.x. Epub 2007 Oct 23.
This study describes the development of a real-time PCR protocol for rapid detection of the most common bla(VIM) (bla(VIM-1), bla(VIM-2), bla(VIM-3), bla(VIM-4), bla(VIM-5), bla(VIM-6), bla(VIM-10), bla(VIM-11), bla(VIM-12)) and bla(IMP) (bla(IMP-1), bla(IMP-2), bla(IMP-6), bla(IMP-8), bla(IMP-10), bla(IMP-15), bla(IMP-19), bla(IMP-20)) genes in a single reaction. The genes were specifically detected and clearly differentiated into four groups, i.e., (i) bla(VIM-1)-like (bla(VIM-1), bla(VIM-4), bla(VIM-5), bla(VIM-12)); (ii) bla(VIM-2)-like (bla(VIM-2), bla(VIM-3), bla(VIM-6), bla(VIM-10), bla(VIM-11)); (iii) bla(IMP-1)-like (bla(IMP-1), bla(IMP-6), bla(IMP-10)); and (iv) bla(IMP-2)-like (bla(IMP-2), bla(IMP-8), bla(IMP-15), bla(IMP-19), bla(IMP-20)), by melting curve analysis of the real-time PCR products. The protocol was used to screen positive bla(VIM-1), bla(VIM-2) and bla(IMP-1) control strains, 70 Gram-negative isolates resistant to carbapenems, and 30 Gram-negative isolates susceptible to carbapenems (negative controls).
本研究描述了一种实时荧光定量PCR方法的开发，该方法可在单一反应中快速检测最常见的bla(VIM)基因（bla(VIM-1)、bla(VIM-2)、bla(VIM-3)、bla(VIM-4)、bla(VIM-5)、bla(VIM-6)、bla(VIM-10)、bla(VIM-11)、bla(VIM-12)）和bla(IMP)基因（bla(IMP-1)、bla(IMP-2)、bla(IMP-6)、bla(IMP-8)、bla(IMP-10)、bla(IMP-15)、bla(IMP-19)、bla(IMP-20)）。通过对实时荧光定量PCR产物进行熔解曲线分析，可特异性检测这些基因，并将其明确分为四组，即：(i) bla(VIM-1)样（bla(VIM-1)、bla(VIM-4)、bla(VIM-5)、bla(VIM-12)）；(ii) bla(VIM-2)样（bla(VIM-2)、bla(VIM-3)、bla(VIM-6)、bla(VIM-10)、bla(VIM-11)）；(iii) bla(IMP-1)样（bla(IMP-1)、bla(IMP-6)、bla(IMP-10)）；以及(iv) bla(IMP-2)样（bla(IMP-2)、bla(IMP-8)、bla(IMP-15)、bla(IMP-19)、bla(IMP-20)）。该方法用于筛选bla(VIM-1)、bla(VIM-2)和bla(IMP-1)阳性对照菌株、70株对碳青霉烯类耐药的革兰氏阴性分离株以及30株对碳青霉烯类敏感的革兰氏阴性分离株（阴性对照）。
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