Real-Time Detection of blaKPC in Clinical

A real-time PCR assay was developed targeting the blaKPC responsible for Klebsiella pneumoniae carbapen-emase (KPC)-mediated carbapenem resistance and was validated for testing colonies or enrichment broth cultures. The assay accurately detects KPC-containing strains with high analytical specificity and sensitivity. Broad-spectrum -lactamases, such as Klebsiella pneumoniae carbapenemases (KPCs), are spreading widely within the United States and cause resistance to virtually all beta-lactam antimicrobials (4, 5). Rapid identification of KPC-containing Enterobacteriaceae is critical to controlling the spread of these pathogens and can be accomplished by molecular detection (6). We have developed and validated a real-time PCR assay for blaKPC detection, including an internal control, and applied it to various clinical specimens, including isolated colonies and surveillance swabs from intensive care unit (ICU) patients after broth enrichment, as a tool for infection control of KPC resistance. For analytical validation, 46 KPC-positive isolates were cul-tured on MacConkey agar (Remel, Thermo Fisher Scientific, Lenexa, KS) plates containing 4 g/ml meropenem (AstraZenec