Application of polymerase chain reaction and denaturing gradient gel electrophoresis assay of the flagellin gene for direct detection and subtyping of Campylobacter jejuni and Campylobacter coli in avian faecal samples
2012
Vashin, I., Thracian University, Stara Zagora (Bulgaria) | Stoyanchev, T., Thracian University, Stara Zagora (Bulgaria) | Iliev, m., Bulgarian Academy of Sciences, Sofia (Bulgaria) | Naydenski, H., Bulgarian Academy of Sciences, Sofia (Bulgaria)
The present report investigated the potential of polymerase chain reaction- PCR and denaturing gradient gel electrophoresis assay of the flagellin gene - fla-DGGE for direct detection, identification and typing of Campylobacter spp., and pathogenic Campylobacter jejuni and Campylobacter сoli in samples without cultivation. Faecal samples obtained from synanthropic bird species inhabiting areas around broiler chickens fattening farms, and wild-type C. jejuni strains isolated from a poultry slaughterhouse were assayed. The PCR assay for Campylobacter spp. and the specific PCR assays for C. jejuni and C. сoli generated fragments sized 816, 735 and 500 bp, respectively. Out of the 34 faecal samples studied, 13 were positive for Campylobacter spp. C. jejuni was detected in 10 - 76.9% and C. сoli – only in one - 7.69% out of Campylobacter positive samples. The employed fla-DGGE was applicable in faecal DNA concentrations over 150 ng/mL. Common genotypes with similar fla-DGGE profiles were found out in both faecal samples from synanthropic birds and broiler chicken isolates.
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