Rapid detection of the Klebsiella pneumoniae carbapenemase (KPC) gene by loop-mediated isothermal amplification (LAMP)

被引：50

作者：

Nakano, Ryuichi ^{[1
]}

Nakano, Akiyo ^{[1
]}

Ishii, Yoshikazu ^{[2
]}

Ubagai, Tsuneyuki ^{[1
]}

Kikuchi-Ueda, Takane ^{[1
]}

Kikuchi, Hirotoshi ^{[1
]}

Tansho-Nagakawa, Shigeru ^{[1
]}

Kamoshida, Go ^{[1
]}

Mu, Xiaoqin ^{[1
,3
]}

Ono, Yasuo ^{[1
]}

机构：

[1] Teikyo Univ, Sch Med, Dept Microbiol & Immunol, Itabashi Ku, Tokyo 1738605, Japan

[2] Toho Univ, Sch Med, Dept Microbiol & Infect Dis, Ota Ku, Tokyo 1438540, Japan

[3] Genom Res Ctr, State Prov Key Labs Biomed Pharmaceut China 1, Harbin 150081, Heilongjiang Pr, Peoples R China

来源：

JOURNAL OF INFECTION AND CHEMOTHERAPY | 2015年 / 21卷 / 3-4期

关键词：

Antibiotic resistance gene; Klebsiella pneumoniae carbapenemase (KPC); Loop-mediated isothermal amplification; Rapid detection; GRAM-NEGATIVE BACTERIA; REAL-TIME PCR; BETA-LACTAMASE; ESCHERICHIA-COLI; NEW-YORK; 1ST REPORT; ENTEROBACTERIACEAE; RESISTANCE; EPIDEMIOLOGY; BLA(KPC);

D O I：

10.1016/j.jiac.2014.11.010

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

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 degrees C, and no cross-eactivity was observed for other types of beta-lactamase (class A, B, C, or D) producers. The detection limit for this assay was found to be 10(0) CFU per tube, in 25 mm, 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. (C) 2014, Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

引用

页码：202 / 206

页数：5

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