Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Campylobacter jejuni and Campylobacter coli.
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 x 10(3) c.f.u. g(-1) (1.4 c.f.u. per test tube) and 4.8 x 10(3) 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.
Yamazaki
Wataru
W
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Taguchi
Masumi
M
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Ishibashi
Masanori
M
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Kitazato
Miyoshi
M
Yodogawa Christian Hospital, Osaka, Japan.
Nukina
Masafumi
M
Microbiological Department, Kobe Institute of Health, Hyogo, Japan.
Misawa
Naoaki
N
Department of Veterinary Public Health, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Inoue
Kiyoshi
K
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
D002169:Campylobacter Infections / Q000175:diagnosis* / Q000382:microbiology; D017000:Campylobacter coli / Q000235:genetics / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology*; D005517:Foodborne Diseases / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D004269:DNA, Bacterial