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Nakano R.,Teikyo University | Nakano A.,Teikyo University | Ishii Y.,Toho University | Ubagai T.,Teikyo University | And 7 more authors.
Journal of Infection and Chemotherapy | Year: 2015

Klebsiella pneumoniae carbapenemases (KPC), which are associated with resistance to carbapenem, have recently spread worldwide and have become a global concern. It is necessary to detect KPC-producing organisms in clinical settings to be able to control the spread of this resistance. We have developed a loop-mediated isothermal amplification (LAMP) method for rapid detection of KPC producers. LAMP primer sets were designed to recognize the homologous regions of bla KPC-2 to bla KPC-17 and could amplify bla KPC rapidly. The specificity and sensitivity of the primers in the LAMP reactions for bla KPC detection were determined. This LAMP assay was able to specifically detect KPC producers at 68 °C, and no cross-reactivity was observed for other types of β-lactamase (class A, B, C, or D) producers. The detection limit for this assay was found to be 100 CFU per tube, in 25 min, which was 10-fold more sensitive than a PCR assay for bla KPC detection. Then, the sensitivity of the LAMP reactions for bla KPC detection in human specimens (sputum samples, urine samples, fecal samples and blood samples) was analyzed; it was observed that the LAMP assay had almost the same sensitivity in these samples as when using purified DNA. The LAMP assay is easy to perform and rapid. It may therefore be routinely applied for detection of KPC producers in the clinical laboratory. © 2014 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases.

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