Open AccessComparison of Loop-Mediated Isothermal Amplification Assay and Conventional Culture Methods for Detection of Campylobacter jejuni and Campylobacter coli in Naturally Contaminated Chicken Meat Samples
Author(s) -
Wataru Yamazaki,
Masumi Taguchi,
Takao Kawai,
Kentaro Kawatsu,
Junko Sakata,
Kiyoshi Inoue,
Naoaki Misawa
Publication year - 2009
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.02004-08
Subject(s) - campylobacter jejuni , loop mediated isothermal amplification , microbiology and biotechnology , campylobacter , biology , enrichment culture , escherichia coli , contamination , enterobacteriaceae , isolation (microbiology) , chromatography , food science , bacteria , chemistry , dna , ecology , biochemistry , genetics , gene
We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for detection of chicken meat samples naturally contaminated withCampylobacter jejuni andCampylobacter coli . A total of 144 Preston enrichment broth cultures from chicken meat samples were assessed by using the LAMP assay and conventional culture methods, which consist of a combination of Preston enrichment culturing and plating onto Butzler and modified charcoal cefoperazone deoxycholate agars. Compared withC. jejuni -C. coli isolation using the conventional culture test, the LAMP results showed 98.5% (67/68) and 97.4% (74/76) sensitivity and specificity, respectively, and the positive and negative predictive values were 97.1% (67/69) and 98.7% (74/75), respectively. The conventional culture test required more than 3 to 4 days to isolate and identifyC. jejuni andC. coli in the Preston enrichment cultures. In contrast, the LAMP assay was markedly faster, requiring less than 90 min from the beginning of DNA extraction to final detection and differentiation ofC. jejuni andC. coli . In total, the LAMP assay required 23.5 to 25.5 h from the beginning of the enrichment culture to final determination. These results suggest that our LAMP assay is a powerful tool for rapid, sensitive, and practical detection ofC. jejuni andC. coli which may facilitate surveillance and control ofC. jejuni-C. coli contamination in chicken, as well as investigations of food poisoning incidents caused by these organisms. This is the first report of a highly sensitive and specific LAMP assay to detect and differentiateC. jejuni andC. coli in chicken meat samples.