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Logo of anclinmicBioMed CentralBiomed Central Web Sitesearchsubmit a manuscriptregisterthis articleAnnals of Clinical Microbiology and Antimicrobials
 
Ann Clin Microbiol Antimicrob. 2012; 11: 9.
Published online 2012 June 18. doi:  10.1186/1476-0711-11-9
PMCID: PMC3377543

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

Abstract

Background

Klebsiella pneumonia 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.

Methods

Real-time PCR assay based on SYBR GreenIwas designed to amplify a 106bp product of the blaKPC 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.

Results

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.

Conclusion

The real-time PCR assay could rapidly and accurately detect KPCs -harboring strains with high analytical sensitivity and specificity.

Keywords: Real-time polymerase chain reaction, Klebsiella pneumonia carbapenemase

Articles from Annals of Clinical Microbiology and Antimicrobials are provided here courtesy of BioMed Central