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Open Access Research

Rapid and sensitive detection of bla KPC gene in clinical isolates of Klebsiella pneumoniae by a molecular real-time assay

Adriana Mosca1, Luisa Miragliotta1, Raffaele Del Prete1, Gerasimos Tzakis1, Lidia Dalfino2, Francesco Bruno2, Laura Pagani3, Roberta Migliavacca3, Aurora Piazza3 and Giuseppe Miragliotta1*

Author Affiliations

1 Department of Interdisciplinary Medicine Microbiology, University of Bari, Piazza Giulio Cesare, Bari, 70124, Italy

2 Department of Emergency and Organ Transplantation Anesthesia and Intensive Care Unit, University of Bari, Piazza Giulio Cesare, Bari, 70124, Italy

3 Microbiology, University of Pavia, via Brambilla 74, Pavia, 27100, Italy

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SpringerPlus 2013, 2:31  doi:10.1186/2193-1801-2-31

Published: 30 January 2013

Abstract

Background

The aim of this study was the rapid identification of blaKPC gene in 38 Klebsiella pneumoniae clinical isolates with reduced susceptibility to carbapenems. The modified Hodge Test (MHT) was carried out to phenotypically determine whether resistance to carbapenems was mediated by a carbapenemase. The detection of the blaKPC gene was performed by real-time acid nucleic sequence-based amplification (NASBA™™), specifically designed for the detection of KPC RNA target.

Results

Thirty-two/38 isolates evaluated by MHT showed the production of carbapenemases, while all the strains exhibited the production of KPC by inhibition test with phenylboronic acid (the combined disk test with IPM/IPM plus phenylboronic acid). The detection of blaKPC gene by Nuclisens EasyQ KPC yielded positive results in 38/38 (100%) strains. The presence of blaKPC gene was confirmed in all K. pneumoniae isolates when tested by the gold standard PCR assay.

Conclusions

In consideration of the serious challenge represented by infections due to K. pneumoniae it appears necessary the rapid identification of carbapenemases in clinical settings as it is made possible by the use of NASBA™ assay.

Keywords:
Klebsiella pneumoniae; Carbapenem resistance; blaKPC; NASBA™