Comparison of Loop-Mediated Isothermal Amplification Assay and Conventional Culture Methods for Detection of Campylobacter jejuni and Campylobacter coli in Naturally Contaminated Chicken Meat Samples

Abstract

We investigated the efficacy of a loop-mediated isothermal amplification (LAMP) assay for detection of chicken meat samples naturally contaminated with Campylobacter jejuni and Campylobacter 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 with C. 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 identify C. jejuni and C. 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 of C. jejuni and C. 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 of C. jejuni and C. coli which may facilitate surveillance and control of C. 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 differentiate C. jejuni and C. coli in chicken meat samples.

Prevalence and Characteristics of <i>Salmonella</i> and <i>Campylobacter</i> in Retail Poultry Meat in Japan

“…enterica and Campylobacter spp., respectively. These frequencies were consistent with those reported in some previous studies (14,15), but higher than those in other reports (5,(16)(17)(18)(19). Several factors might influence the prevalence of Salmonella and Campylobacter isolates in poultry meat: environmental factors, such as the geographical location of farms and the season in which the study was carried out, as well as differences in bacterial culture conditions and sampling methods (20).…”

Loop-Mediated Isothermal Amplification (LAMP) for Detection of <i>Campylobacter jejuni</i> and <i>C. coli</i> in Thai Children with Diarrhea

“…Conventional culture testing is recognized as the gold standard for the diagnosis of Campylobacter diarrheal disease. This method, however, is complex and time-consuming and may fail to detect these microorganisms (4). Rapid and accurate diagnosis of Campylobacter infection in infants and children is necessary for timely (early) treatment.…”

Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a <i>Campylobacter jejuni</i> Clone

“…A number of LAMP assays for Campylobacter have previously been developed for detection of different Campylobacter species in various sources including human diarrheal stool [16] and chicken meat samples [18], and for differentiation of closely related Campylobacter subspecies in pure culture [17,19]. A common conclusion from these studies is that the LAMP assay was faster and simpler than regular PcR and/or culture while being as sensitive and specific for the purpose of use and that it was thus a practical tool for surveillance and other epidemiological studies.…”

“…Despite being effective and accurate, the bacterial culture and biochemical assays may require up to 4 days and thus are time-consuming. PCR-based assays are more rapid to identify Campylobacter species than bacterial culture and biochemical assays, but the requirement of sophisticated procedures and expensive equipments to detect the DNA targets may greatly restrict their application in clinical settings.Recently, a rapid and simple assay named loop-mediated isothermal amplification (LAMP) [6] for the detection of C. jejuni [16,18], Campylobacter coli [16,18] and Campylobacter fetus [17,19] has been developed. The LAMP assay is a highly specific strand displacement amplification method, using 4 to 6 different primers designed to 8 from 6 distinct regions of the target DNA sequence.…”

“…Recently, a rapid and simple assay named loop-mediated isothermal amplification (LAMP) [6] for the detection of C. jejuni [16,18], Campylobacter coli [16,18] and Campylobacter fetus [17,19] has been developed. The LAMP assay is a highly specific strand displacement amplification method, using 4 to 6 different primers designed to 8 from 6 distinct regions of the target DNA sequence.…”