Characterization of Campylobacter jejuni and Campylobacter coli genotypes in poultry flocks by restriction fragment length polymorphism (RFLP) analysis.
We describe a simple, rapid, and discriminatory methodology that allows the routine molecular characterization of Campylobacter jejuni and Campylobacter coli isolates. The proposed approach is built on one of the earliest and simplest molecular typing methods ever, consisting on the analysis of the fragments of different lengths generated by digestion of homologous DNA sequences with specific restriction endonucleases, a process known as restriction fragment length polymorphism (RFLP) analysis. The strategy underneath the workflow reported here is meant to explore the polymorphisms of Campylobacter spp. flaA gene (flaA-RFLP) that allows the local investigation of the genetic diversity and distribution of C. coli and C. jejuni isolates from different sources, namely, chickens' caeca. Although not appropriate for global and long-term epidemiological studies as a single approach, flaA-RFLP analysis can be very useful in surveys limited in space and time and, for specific epidemiological settings, an alternative to more modern and resource-demanding techniques.
iMed.UL-Research Institute for Medicines and Pharmaceutical Sciences, Intracellular Trafficking Modulation for Advanced Drug Delivery, Faculdade de Farmácia, Universidade de Lisboa, Avenida Prof. Gama Pinto, 1649-003, Lisboa, Portugal.
Ana Cláudia
AC
Carreira
Mónica V
MV
Cunha
D000818:Animals; D002169:Campylobacter Infections / Q000662:veterinary*; D017000:Campylobacter coli / Q000235:genetics*; D016123:Campylobacter jejuni / Q000235:genetics*; D005838:Genotype; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology*