Naserpour Farivar Taghi, Najafipour Reza, Johari Pouran, Aslanimehr Masoumeh, Peymani Amir, Jahani Hashemi Hoasan, Mirzaui Baman
Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
Iran J Microbiol. 2014 Oct;6(5):335-40.
We developed and evaluated the utility of a quadruplex Taqman real-time PCR assay that allows simultaneous identification of vancomycin-resistant genotypes and clinically relevant enterococci.
The specificity of the assay was tested using reference strains of vancomycin-resistant and susceptible enterococci. In total, 193 clinical isolates were identified and subsequently genotyped using a Quadruplex Taqman real-time PCR assay and melting curve analysis. Representative Quadruplex Taqman real-time PCR amplification curve were obtained for Enterococcus faecium, Enterococcus faecalis, vanA-containing E. faecium, vanB-containing E. faecalis.
Phenotypic and genotypic analysis of the isolates gave same results for 82 enterococcal isolates, while in 5 isolates, they were inconsistent. We had three mixed strains, which were detected by the TaqMan real-time PCR assay and could not be identified correctly using phenotypic methods.
Vancomycin resistant enterococci (VRE) genotyping and identification of clinically relevant enterococci were rapidly and correctly performed using TaqMan real-time multiplex real-time PCR assay.
我们开发并评估了一种四重Taqman实时PCR检测方法的实用性，该方法可同时鉴定耐万古霉素基因型和临床相关肠球菌。
使用耐万古霉素和敏感肠球菌的参考菌株测试该检测方法的特异性。总共鉴定了193株临床分离株，随后使用四重Taqman实时PCR检测方法和熔解曲线分析对其进行基因分型。获得了粪肠球菌、屎肠球菌、含vanA的屎肠球菌、含vanB的粪肠球菌的代表性四重Taqman实时PCR扩增曲线。
对分离株的表型和基因型分析显示，82株肠球菌分离株的结果相同，而5株分离株的结果不一致。我们有三株混合菌株，通过TaqMan实时PCR检测方法检测到，使用表型方法无法正确鉴定。
使用TaqMan实时多重PCR检测方法可快速、正确地对耐万古霉素肠球菌（VRE）进行基因分型并鉴定临床相关肠球菌。
