The establishment of a duplex real-time PCR assay for rapid and simultaneous detection of blaNDM and blaKPC genes in bacteria

Annals of Clinical Microbiology and Antimicrobials

Research

Yongyu Rui¹  Fen Zheng¹  Jingjing Sun¹  Cancan Cheng¹ 

[1] Laboratory Medicine Center, Nanfang Hospital, Southern Medical University, 510515, Guangzhou, China;

关键词: Duplex;    Real-time PCR;    Infection control;    Carbapenemase;   

DOI  :  10.1186/1476-0711-12-30

 received in 2013-09-05, accepted in 2013-10-14,  发布年份 2013

来源: Springer

【 摘 要 】

The latest threat of multidrug-resistant Gram-negative bacteria corresponds to the emergence of carbapenemase New Delhi metallo-β-lactamase (NDM) and Klebsiella pneumoniae carbapenemase (KPC) producers. Rapid molecular detection is essential to limit their spread. In this study, a duplex real-time polymerase chain reaction (PCR) that was specific for the detection of blaNDM and bl aKPC with the same limit of detection of ten plasmid copies was developed. The assay was linear over eight log dilutions for blaNDM (R2 = 0.971; slope, -3.273) and blaKPC (R2 = 0.992; slope, -2.997) with efficiencies of 102% and 115%, respectively. The assay was validated with 157 clinical isolates and showed 100% concordance with conventional PCR. The excellent performance of the duplex PCR assay makes it a powerful tool for surveillance of the carbapenemases NDM and KPC.

【 授权许可】

Unknown   
© Zheng et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.