Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Campylobacter jejuni and Campylobacter coli

Author:

Yamazaki Wataru1,Taguchi Masumi1,Ishibashi Masanori1,Kitazato Miyoshi2,Nukina Masafumi3,Misawa Naoaki4,Inoue Kiyoshi1

Affiliation:

1. Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan

2. Yodogawa Christian Hospital, Osaka, Japan

3. Microbiological Department, Kobe Institute of Health, Hyogo, Japan

4. Department of Veterinary Public Health, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan

Abstract

We developed a loop-mediated isothermal amplification (LAMP) assay for the rapid and simple detection of Campylobacter jejuni and Campylobacter coli. The assay provides a specific LAMP product for each of these two species. The assay correctly identified 65 C. jejuni and 45 C. coli strains, but not 75 non-C. jejuni/coli strains. The sensitivity of the LAMP assay for C. jejuni and C. coli in spiked human stool specimens was 5.6×103 c.f.u. g−1 (1.4 c.f.u. per test tube) and 4.8×103 c.f.u. g−1 (1.2 c.f.u. per test tube), respectively. When 90 stool specimens from patients with diarrhoea were tested by LAMP and direct plating, the LAMP results showed 81.3 % sensitivity and 96.6 % specificity compared to isolation of C. jejuni and C. coli by direct plating. Further, the LAMP assay required less than 2 h for detection of C. jejuni and C. coli in stool specimens. This LAMP assay is a rapid and simple tool for the detection of C. jejuni and C. coli and will be useful in facilitating the early diagnosis of food poisoning incidents caused by these organisms.

Publisher

Microbiology Society

Subject

Microbiology (medical),General Medicine,Microbiology

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