Evaluation of a Real-Time PCR Assay for the Detection of the Klebsiella pneumoniae Carbapenemase Genes in Microbiological Samples in Comparison With the Modified Hodge Test | Zendy

Aditya Raghunathan | Zendy; Linoj Samuel | Zendy; Robert Tibbetts | Zendy

Open Access

Evaluation of a Real-Time PCR Assay for the Detection of the Klebsiella pneumoniae Carbapenemase Genes in Microbiological Samples in Comparison With the Modified Hodge Test

Author(s) -

Aditya Raghunathan,

Linoj Samuel,

Robert Tibbetts

Publication year - 2011

Publication title -

american journal of clinical pathology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.859

H-Index - 128

eISSN - 1943-7722

pISSN - 0002-9173

DOI - 10.1309/ajcppvnui3o9jhjw

Subject(s) - klebsiella pneumoniae , polymerase chain reaction , biology , microbiology and biotechnology , real time polymerase chain reaction , primer (cosmetics) , sybr green i , gene , genetics , escherichia coli , chemistry , organic chemistry

Transfer of the bla(KPC) genes encoding the Klebsiella pneumoniae carbapenemase (KPC) are increasingly responsible for emerging carbapenem resistance. The modified Hodge test (MHT) is recommended for the detection of KPC. We compared MHT with a real-time polymerase chain reaction (PCR) assay targeting common subtypes of bla(KPC), using previously described forward and reverse primer sequences. The PCR product was detected using SYBR Green (Applied Biosystems, Foster City, CA) and confirmed by melt curve analysis. PCR was positive in 96% (52/54) of isolates that were MHT+, 90% (28/31) of MHT- isolates were PCR-, and the results were strongly correlated (P = .0001; Fisher exact test). The PCR assay is a sensitive, specific, and rapid test for detecting bla(KPC) genes. It could help optimize patient care by reducing the time taken to institute appropriate antimicrobial therapy and so help improve patient outcomes.

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