Abstract
Campylobacter jejuni and Campylobacter coli are common causes of gastroenteritis in humans. Infection with C. jejuni or C. coli is commonly acquired by eating undercooked chicken. The goal of this study was to develop specific detection assays for C. jejuni and C. coli isolates based on the cadF virulence gene and its product. The cadF gene from C. jejuni and C. coli encodes a 37-kDa outer membrane protein that promotes the binding of these pathogens to intestinal epithelial cells. A fragment of approximately 400 bp was amplified from 38 of 40 (95%) C. jejuni isolates and 5 of 6 (83.3%) C. coli isolates with primers designed to amplify an internal fragment of the cadF gene. PCR was then used to amplify Campylobacter DNA from store-bought chickens. A 400-bp band was amplified from 26 of the 27 chicken carcasses tested by the PCR-based assay. The CadF protein was detected in every C. jejuni and C. coli isolate tested, as judged by immunoblot analysis with a rabbit anti-C. jejuni 37-kDa serum. In addition, methanol-fixed samples of whole-cell C. jejuni and C. coli were detected with the rabbit anti-37-kDa serum by using an indirect-immunofluorescence microscopy assay. These findings indicate that the cadF gene and its product are conserved among C. jejuni and C. coli isolates and that a PCR assay based on the cadF gene may be useful for the detection of Campylobacter organisms in food products.
MeSH Terms
Animals
Bacterial Outer Membrane Proteins/genetics
Base Sequence
Campylobacter Infections/etiology,microbiology
Campylobacter coli/classification,genetics,pathogenicity
Campylobacter jejuni/classification,genetics,pathogenicity
Carrier Proteins/genetics
Chickens/microbiology
Gastroenteritis/etiology,microbiology
Humans
Meat/microbiology
Molecular Sequence Data
Polymerase Chain Reaction
Polymorphism, Genetic
Rabbits
Sensitivity and Specificity
Sequence Alignment
Sequence Homology, Nucleic Acid
Serotyping
Virulence/genetics
Chemicals
Bacterial Outer Membrane Proteins
CadF protein, Campylobacter jejuni
Carrier Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Konkel M E
Department of Microbiology, Washington State University, Pullman, Washington 99164-4233, USA.
[email protected]
Gray S A
Kim B J
Garvis S G
Yoon J
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