Direct polymerase chain reaction detection of Campylobacter jejuni and Campylobacter coli in raw milk and dairy products

Abstract

A polymerase chain reaction (PCR) method designed to sensitively detect and identify Campylobacterjejuni and Campylobacter coli without the need for isolating and culturing strains is described. The intergenic sequence between the flageilin genesfla& andflaB was amplified and characterized with a triple primer or seminested primer approach. A total of 50 bacterial strains, 27 of C. jejuni and C. coli and 23 of other species, were tested, giving no false-positive or false-negative results. The detection limit as determined by ethidium bromide staining of amplification products on agarose gels was 10 bacteria or less in artificially contaminated water, milk, and soft cheese samples with the seminested primer PCR assay. As an application of the PCR system, a set of 93 samples of milk and other dairy products was screened for the presence of C. jejuni and C. coli. We identified six positive samples (6.5%), while none were found with a conventional culture method.

Campylobacter jejuni prevalence and hygienic quality of retail bovine ground meat in Finland

“…jejuni from foodstuffs using direct PCR. However, a study by Wegm€ uller et al (1993) developed a seminested PCR protocol that targeted the intragenic flaA-flaB region and used it to test cheese, soft-cheese and milk samples. The detection limit of their method was low: <10 bacterial cells per 2 g of matrix (Wegm€ uller et al 1993).…”

“…However, a study by Wegm€ uller et al (1993) developed a seminested PCR protocol that targeted the intragenic flaA-flaB region and used it to test cheese, soft-cheese and milk samples. The detection limit of their method was low: <10 bacterial cells per 2 g of matrix (Wegm€ uller et al 1993). Another study used the same method on a template extracted directly from four inoculated ground meat samples and even though they reported a lower detection limit compared to our PCR-D method, the results were not reproducible (Waage et al 1999).…”

A PCR assay for the detection of Campylobacter jejuni and Campylobacter coli in water

“…coli, being able to discriminate between these species and C a m p . lari their closest relative (Wegmuller et al 1993).…”

“…Oligonucleotide primers CF02, CF03 and CF04 (Wegmuller et al 1993) Five experimental runs were carried out with both C a m p . jejuni and C a m p .…”

A MULTIPLEX POLYMERASE CHAIN REACTION ASSAY FOR RAPID DETECTION OF ESCHERICHIA COLI O157:H7, LISTERIA MONOCYTOGENES, SALMONELLATYPHIMURIUM AND CAMPYLOBACTER JEJUNI IN ARTIFICIALLY CONTAMINATED FOOD SAMPLES

“…Polymerase chain reaction (PCR) assays have been used in the rapid and specific detection of each of these pathogens separately for more than two decades (Ng et al 1997;Winter et al 1997Winter et al , 1999Wang et al 1999;Holko et al 2002;Grant 2003). PCR methods are also used in the detection of food-borne bacterial pathogens from contaminated foods as part of a quality-control system of food safety (Wegmuller et al 1993;Wang et al 1999;Waller and Ogata 2000). The detection of these pathogens in contaminated foods by a single assay would be very useful because it would be rapid and cost-effective.…”