Application of a loop-mediated isothermal amplification method for the detection of pathogenic Leptospira

被引：53

作者：

Lin, Xu'ai ^{[1
]}

Chen, Yin ^{[2
]}

Lu, Yiyu ^{[2
]}

Yan, Juying ^{[2
]}

Yan, Jie ^{[1
]}

机构：

[1] Zhejiang Univ, Dept Med Microbiol & Parasitol, Sch Med, Hangzhou 310058, Zhejiang, Peoples R China

[2] Zhejiang Prov Ctr Dis Control & Prevent, Hangzhou 310009, Zhejiang, Peoples R China

来源：

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE | 2009年 / 63卷 / 03期

关键词：

Loop-mediated isothermal amplification; Leptospira; Detection; Real-time; PCR; REAL-TIME PCR; QUANTITATIVE PCR; DIAGNOSIS; TUBERCULOSIS; SAMPLES; DNA;

D O I：

10.1016/j.diagmicrobio.2008.10.012

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Leptospirosis is an emerging inflections disease, which is considered to be the most widespread zoonotic disease in the world. There are more than 230 known serovars in the genus Leptospira. A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of pathogenic Leptospira spp. was developed and evaluated through amplification of the lipL41 gene coding for the Outer membrane protein LipL41. The LAMP assay did not rely oil the isolation and Culture of leptospires, and no cross-reactivity was observed with other bacterial species. A SYBR Green I-based LAMP assay was also carried out for the real-time detection of DNA amplification. The lower detection limit of the LAMP assay was approximately 100 copies, which was the same as the polymerase chain reaction (PCR) and real-time PCR assays. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis of the amplified product. The LAMP assay is easy to perform and inexpensive, and so may be applied in the rapid and specific diagnosis of Leptospira. (c) 2009 Elsevier Inc. All rights reserved.

引用

页码：237 / 242

页数：6