Identification of the enteropathogens Campylobacter jejuni and Campylobacter coli based on the cadF virulence gene and its product.
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.
Konkel
M E
ME
Department of Microbiology, Washington State University, Pullman, Washington 99164-4233, USA. konkel@mail.wsu.edu
Gray
S A
SA
Kim
B J
BJ
Garvis
S G
SG
Yoon
J
J
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D002169:Campylobacter Infections / Q000209:etiology / Q000382:microbiology; D017000:Campylobacter coli / Q000145:classification / Q000235:genetics / Q000472:pathogenicity*; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics / Q000472:pathogenicity*; D002352:Carrier Proteins / Q000235:genetics*; D002645:Chickens / Q000382:microbiology; D005759:Gastroenteritis / Q000209:etiology / Q000382:microbiology; D006801:Humans; D008460:Meat / Q000382:microbiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011817:Rabbits; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D012703:Serotyping; D014774:Virulence / Q000235:genetics