Rapid detection of Pseudomonas aeruginosa in mouse feces by colorimetric loop-mediated isothermal amplification

被引：60

作者：

Goto, Motoki ^{[2
]}

Shimada, Kayo ^{[3
]}

Sato, Ayako ^{[1
]}

Takahashi, Eri ^{[1
]}

Fukasawa, Takafumi ^{[1
]}

Takahashi, Tomoki ^{[1
]}

Ohka, Seii ^{[2
]}

Taniguchi, Takahide ^{[4
]}

Honda, Eiichi ^{[4
]}

Nomoto, Akio ^{[2
]}

Ogura, Atsuo ^{[2
,5
]}

Kirikae, Teruo ^{[3
]}

Hanaki, Ken-Ichi ^{[1
]}

机构：

[1] Iwate Med Univ, Ctr Expt Anim Res, Morioka, Iwate 0208505, Japan

[2] Univ Tokyo, Grad Sch Med, Ctr Dis Biol & Integrat Med, Bunkyo Ku, Tokyo 1130033, Japan

[3] Int Med Ctr Japan, Res Inst, Dept Infect Dis, Shinjuku Ku, Tokyo 1628655, Japan

[4] Tokyo Univ Agr & Technol, Dept Vet Med, Fuchu, Tokyo 1838509, Japan

[5] RIKEN BioResource Ctr, Tsukuba, Ibaraki 3050074, Japan

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2010年 / 81卷 / 03期

关键词：

Hydroxy naphthol blue; Loop-mediated isothermal amplification; OprL gene; Pseudomonas aeruginosa; CYSTIC-FIBROSIS PATIENTS; POLYMERASE-CHAIN-REACTION; CLINICAL-SAMPLES; PCR; MICE; IDENTIFICATION; INFECTION; VIRUS; RELIABILITY; BACTEREMIA;

D O I：

10.1016/j.mimet.2010.03.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A colorimetric loop-mediated isothermal amplification (LAMP) assay with hydroxy naphthol blue was designed to amplify a region in the outer membrane lipoprotein (oprL) gene of Pseudomonas aeruginosa. The LAMP assay showed 100% specificity for the serogroup and other bacteria, and the sensitivity was 10-fold higher than that of the PCR assays. The LAMP assay could detect P. aeruginosa inoculated in mouse feces at 130 colony-forming units (CFU)/0.1 g feces (3.25 CFU/reaction). The assay was completed within 2 h from DNA extraction. In a field trial, the LAMP assay revealed that none of the 27 samples was obtained from 2 specific pathogen-free (SPF) mouse facilities that were monitoring infection with P. aeruginosa; 1 out of 12 samples from an SPF mouse facility that was not monitoring infection with P. aeruginosa and 2 out of 7 samples from a conventional mouse facility were positive for P. aeruginosa. In contrast, P. aeruginosa was not detected in any of the samples by a conventional culture assay. Thus, this colorimetric LAMP assay is a simple and rapid method for P. aeruginosa detection. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：247 / 252

页数：6

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