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United States Department of Agriculture

Agricultural Research Service

Title: Evaluation of a New Pcr Method for the Identification of Campylobacter Jejuni and Campylobacter Coli

Authors
item Englen, Mark
item Cray, Paula
item Czajka, J - DUPONT QUALICON
item Bolton, L - DUPONT QUALICON
item Peterson, K - USDA-FSIS
item Headrick, Marcia - FDA-CVM
item Tollefson, K - FDA-CVM

Submitted to: American Society for Microbiology
Publication Type: Abstract Only
Publication Acceptance Date: July 7, 2000
Publication Date: N/A

Technical Abstract: Campylobacter is a leading cause of enteritis in humans, often surpassing the number of cases due to Salmonella in the U.S. The two species most frequently isolated from cases of human illness are C. jejuni and C. coli, and poultry and poultry products are often implicated as the sources of these infections. DuPont Qualicon has recently developed a new BAX System PCR assay for the identification of C. jejuni and C. coli and it was evaluated in this study against a PCR method already in use by us (Englen and Fedorka-Cray) for differentiating Campylobacter. The BAX PCR is a multiplex assay in which the primers, dNTPs and Taq DNA polymerase are tableted. Template DNA is prepared by adding cells to a proprietary lysis buffer, carrying out two brief heat steps, then directly adding the lysis mixture to a tube containing the BAX tablet. The standard PCR is not a multiplex, requiring preparation of a master mix of primers, dNTPs and Taq DNA polymerase for each primer set. For template DNA, a suspension of cells in water are boiled, then the DNA is extracted using a commercial reagent (DNAzol; Life Technologies, Gaithersburg, MD). One hundred thirty-seven Campylobacter isolates from poultry carcass rinse samples collected as part of the 1998 National Antimicrobial Resistance Monitoring System (NARMS) survey were screened using the BAX PCR and the results compared to those obtained with the standard PCR. For 119 isolates, identical results were obtained with both assays, yielding 86 C. jejuni and 33 C. coli. The remaining 18 isolates gave conflicting results suggesting mixed cultures, and these were retested by both PCR methods.

Last Modified: 4/23/2014
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