A rapid low-cost real-time PCR for the detection of <it>klebsiella pneumonia</it> carbapenemase genes

• Main Authors: Wang Lijun, Gu Haitong, Lu Xinxin
• Format: Article
• Language: English
• Published: BMC 2012-06-01
• Series: Annals of Clinical Microbiology and Antimicrobials
• Subjects: Real-time polymerase chain reaction; Klebsiella pneumonia carbapenemase

<p>Abstract</p> <p>Background</p> <p><it>Klebsiella pneumonia</it> carbapenemases (KPCs) are able to hydrolyze the carbapenems, which cause many bacteria resistance to multiple classes of antibiotics, so the rapid dissemination of KPCs is worrisome. Laboratory identification of KPCs-harboring clinical isolates would be a key to limit the spread of the bacteria. This study would evaluate a rapid low-cost real-time PCR assay to detect KPCs.</p> <p>Methods</p> <p>Real-time PCR assay based on SYBR GreenIwas designed to amplify a 106bp product of the <it>bla</it>_KPC gene from the159 clinical Gram-negative isolates resistant to several classes of -lactam antibiotics through antimicrobial susceptibility testing. We confirmed the results of real-time PCR assay by the conventional PCR-sequencing. At the same time, KPCs of these clinical isolates were detected by the modified Hodge test (MHT). Then we compared the results of real-time PCR assay with those of MHT from the sensitivity and specificity. Moreover, we evaluated the sensitivity of the real-time PCR assay.</p> <p>Results</p> <p>The sensitivity and specificity of the results of the real-time PCR assay compared with those of MHT was 29/29(100%) and 130/130(100%), respectively. The results of the real-time PCR and the MHT were strongly consistent (Exact Sig. (2-tailed) =1. 000; McNemar test). The real-time PCR detection limit was about 0.8cfu using clinical isolates.</p> <p>Conclusion</p> <p>The real-time PCR assay could rapidly and accurately detect KPCs -harboring strains with high analytical sensitivity and specificity.</p>