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Title: COMPARISON OF TWO MOLECULAR TECHNIQUES FOR EPIDEMIOLOGICAL TRACING OF CAMPYLOBACTER JEJUNI

Author
item CHEN, HUI-CHENG - UGA
item Stern, Norman

Submitted to: Applied and Environmental Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/9/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary: Campylobacter jejuni is the most prominent bacterial cause of human intestinal disease. Poultry is reported as the most important reservoir in its transmission. We wanted to determine an optimum means for distinguishing one Campylobacter strain from another. DNA extracted from different strains was broken down and examined for specific DNA sequences. We could not detect consistent differences between DNA from human or chicken Campylobacter isolates. Using another approach for analysis (flaA-RFLP), we amplified the DNA encoding for the "tails" of Campylobacter, "cut" the DNA up, and created band patterns that were specific for each isolate. We concluded that this second approach was useful for tracing Campylobacter strains. This information can now be applied to further studies monitoring a mixture of Campylobacter strains in chickens. The findings provide an optimal tool for accurate tracking analysis.

Technical Abstract: Two molecular typing methods were compared in this study for capability in distinguishing and characterizing Campylobacter jejuni strains isolated from chickens and humans. Chromosomal DNA fragments were digested with ClaI enzyme and further analyzed by hybridization with a DNA probe, pMO2005 (10, 11). No specific chromosomal DNA patterns were revealed after analyzing 10 chicken and 4 human isolates, and the consistency to generate replicate DNA patterns were also low. Polymerase chain reaction (PCR) amplified flagellin gene (flaA) in C. jejuni were analyzed by restriction fragment length polymorphism (RFLP), digested with DdeI enzyme (16, 17). Various simple patterns, easy-to-read and consistent flaA gene types were generated from 78 chicken and 39 human C. jejuni strains analyzed with RFLP. Our results indicated that RFLP analysis of flaA gene has sufficient discriminating power for epidemiological tracing of C. jejuni strains, while the chromosomal DNA method hybridized with a DNA probe is less discriminatory.