Application of real-time PCR for quantitative detection of Campylobacter jejuni in poultry, milk and environmental water

• Published in: FEMS immunology and medical microbiology 2003-10, Vol.38 (3), p.265-271
• Main Authors: Yang, Chengbo, Jiang, Yuan, Huang, Kehe, Zhu, Changqing, Yin, Yulong
• Format: Article
• Language: English
• Subjects: Animals; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Campylobacter jejuni; Campylobacter jejuni - classification; Campylobacter jejuni - growth & development; Campylobacter jejuni - isolation & purification; Campylobacter jejuni - metabolism; China; Colony Count, Microbial; DNA Primers; DNA, Bacterial - isolation & purification; Food Microbiology; Fundamental and applied biological sciences. Psychology; LightCycler; Meat - microbiology; Microbiology; Milk - microbiology; Polymerase Chain Reaction - methods; Poultry; Quantitative detection; Real-time polymerase chain reaction; Sensitivity and Specificity; TaqMan probe; Water Microbiology
• ISSN: 0928-8244; 1574-695X
• DOI: 10.1016/S0928-8244(03)00168-8

Campylobacter jejuni is a leading human food-borne pathogen. The rapid and sensitive detection of C. jejuni is necessary for the maintenance of a safe food/water supply. In this article, we present a real-time polymerase chain reaction (PCR) assay for quantitative detection of C. jejuni in naturally contaminated poultry, milk and environmental samples without an enrichment step. The whole assay can be completed in 60 min with a detection limit of approximately 1 CFU. The standard curve correlation coefficient for the threshold cycle versus the copy number of initial C. jejuni cells was 0.988. To test the PCR system, a set of 300 frozen chicken meat samples, 300 milk samples and 300 water samples were screened for the presence of C. jejuni. 30.6% (92/300) of chicken meat samples, 27.3% (82/300) of milk samples, and 13.6% (41/300) of water samples tested positive for C. jejuni. This result indicated that the real-time PCR assay provides a specific, sensitive and rapid method for quantitative detection of C. jejuni. Moreover, it is concluded that retail chicken meat, raw milk and environmental water are commonly contaminated with C. jejuni and could serve as a potential risk for consumers in eastern China, especially if proper hygienic and cooking conditions are not maintained.