|Miller, William - Bill|
Submitted to: Journal of Microbiological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/19/2010
Publication Date: 12/2/2010
Citation: Behringer, M., Miller, W.G., Oyarzabal, O.A. 2010. flaA-PCR, MLST, PFGE and Rep-PCR Typing of Campylobacter jejuni and Campylobacter coli isolated from live broilers and retail broiler meat. Journal of Microbiological Methods. 84(2011) 194-201. Interpretive Summary: Campylobacter strains can be typed using any one of (or combinations of) a number of available molecular typing methods. Multiple molecular typing methods exist; however, studies generally compare no more than two of these typing methods on the same strain set. This study tested the four most commonly used methods on a single set of 101 Campylobacter strains isolated from the United States and Grenada. All four typing methods satisfactorily typed C. jejuni and C. coli strains. However, some variables influenced the typing results, for example the extraction method used to isolate the DNA and whether C. jejuni strains, C coli strains or a mixture of both were being typed. Additionally, some strains could not
Technical Abstract: We analyzed a selection of 101 Campylobacter spp. isolates sampled from Grenada, Puerto Rico and Alabama in order to evaluate the discriminatory strength of four prominent molecular fingerprinting methods: restriction fragment length polymorphism of the flaA gene (flaA-RFLP), pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST) and automated repetitive extragenic palindromic polymerase chain reaction using the Diversilab system (Rep-PCR). All performed similarly for the typing of C. jejuni and C. coli. PFGE and MLST had a better discriminatory power for C. coli isolates than Rep-PCR of flaA-RFLP. The DNA extraction method still influences the results obtained with Rep-PCR. This method was better for the typing of C. jejuni than C. coli. Both Rep-PCR and flaA-RFLP generated types that were indistinguishable between C. jejuni and C. coli. PFGE and MLST generated typing results that had a better correlation with the location and the date of collection of isolates than the results obtained with Rep-PCR and flaA-RFLP. Therefore, it is important to have clear epidemiological questions in mind before deciding on the typing technique to incorporate. Further studies using different C. jejuni and C. coli may expand our understanding of the benefits and limitations of each of these typing methods for epidemiological studies of Campylobacter spp. be typed by some of the molecular methods. Finally, some method/strain combinations generated random profiles independent of the epidemiological data. Therefore, the typing method used to discriminate Campylobacter strains depends to some extent on the species being typed, the strains used and the quality of the extracted DNA.