Typing of Campylobacter jejuni and Campylobacter coli isolated from live broilers and retail broiler meat by flaA-RFLP, MLST, PFGE and REP-PCR.
We analyzed 100 Campylobacter spp. isolates (C. jejuni and C. coli) from Grenada, Puerto Rico and Alabama, which were collected from live broilers or retail broiler meat. We analyzed these isolates with four molecular typing methods: restriction fragment length polymorphism of the flaA gene (flaA-RFLP), multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and automated repetitive extragenic palindromic polymerase chain reaction (REP-PCR) using the DiversiLab system. All methods performed similarly for the typing of C. jejuni and C. coli. The DNA extraction method appears to influence the results obtained with REP-PCR. This method was better for the typing of C. jejuni than C. coli, however both REP-PCR and flaA-RFLP generated types that were indistinguishable between C. jejuni and C. coli and appeared to be random, without any relationship to species, location, or source of isolates. PFGE and MLST generated typing results that had a better correlation with the geographic location of the isolates and showed higher concordance with the Wallace coefficient. The adjusted Rand coefficient did not show higher concordance among the methods, although the PFGE/MLST combination exhibited the highest concordance. PFGE and MLST revealed a better discriminatory power for C. coli isolates than REP-PCR or flaA-RFLP. The use of readily available online tools to calculate the confidence interval of the Simpson's index of diversity and the adjusted Rand and Wallace coefficients helped estimate the discriminatory power of typing methods. Further studies using different C. jejuni and C. coli strains may expand our understanding of the benefits and limitations of each of these typing methods for epidemiological studies of Campylobacter spp.
Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University, AL 36849, USA.
Megan
M
Behringer
William G
WG
Miller
Omar A
OA
Oyarzabal
D000407:Alabama; D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods*; D017000:Campylobacter coli / Q000145:classification* / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000145:classification* / Q000302:isolation & purification; D002645:Chickens / Q000382:microbiology*; D016172:DNA Fingerprinting / Q000379:methods; D016521:Electrophoresis, Gel, Pulsed-Field; D005408:Flagellin / Q000235:genetics; D005838:Genotype; D019425:Grenada; D008460:Meat / Q000382:microbiology*; D058889:Molecular Typing / Q000379:methods*; D058885:Multilocus Sequence Typing; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011647:Puerto Rico
D005408:Flagellin; C067138:flaA protein, bacteria