Quantification and differentiation of Campylobacter jejuni and Campylobacter coli in raw chicken meats using a real-time PCR method.
Campylobacter species are one of the most common causes of bacterial diarrhea in humans worldwide. The consumption of foods contaminated with two Campylobacter species, C. jejuni and C. coli, is usually associated with most of the infections in humans. In this study, a rapid, reliable, and sensitive multiplex real-time quantitative PCR was developed for the simultaneous detection, identification, and quantification of C. jejuni and C. coli. In addition, the developed method was applied to the 50 samples of raw chicken meat collected from retail stores in Korea. C. jejuni and C. coli were detected in 88 and 86% of the samples by real-time quantitative PCR and the conventional microbiological method, respectively. The specificity of the primer and probe sets was confirmed with 30 C. jejuni, 20 C. coli, and 35 strains of other microbial species. C. jejuni and C. coli could be detected with high specificity in less than 4 h, with a detection limit of 1 log CFU/ml by the developed real-time PCR. The average counts (log CFU per milliliter) of C. jejuni or C. coli obtained by the conventional methods and by the real-time PCR assay were statistically correlated with a correlation coefficient (R2) between 0.73 and 0.78. The real-time PCR assay developed in this study is useful for screening for the presence and simultaneous differential quantification of C. jejuni and C. coli.
Hong
Joonbae
J
Department of Microbiology, College of Veterinary Medicine and BK21 Program for Veterinary Science, Seoul National University, Sillim-dong, Gwanak-gu, Seoul 151-742, Korea.
Jung
Woo Kyung
WK
Kim
Jun Man
JM
Kim
So Hyun
SH
Koo
Hye Cheong
HC
Ser
Junghee
J
Park
Yong Ho
YH
D000818:Animals; D017000:Campylobacter coli / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000302:isolation & purification*; D002645:Chickens; D015169:Colony Count, Microbial; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D006801:Humans; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018570:Risk Assessment; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013997:Time Factors