DEVELOPMENT OF A MULTIPLEX PCR GENE FINGERPRINTING METHOD USING GYRA AND PFLA POLYMORPHISMS TO IDENTIFY GENOTYPIC RELATEDNESS WITHIN CAMPYLOBACTER-JEJUNI SPECIES

The two genes gyrA and pflA, whose sequence variability had been previ ously described, were evaluated separately for their potential value i n discriminating strains of Campylobacter jejuni. A single method was then developed by which the to loci mere simultaneously amplified usin g a multiplex PCR procedure, and banding patterns were generated using a pre-selected set of restriction endonuclease enzymes. The method wa s applied to 18 strains of Camp. jejuni from different poultry sources varying in geographical origin and year of isolation. Results were co mbined and compared by means of numerical analysis with the classifica tion obtained by flaA-typing and macrorestriction SmaI and KpnI. The u sefulness of PCR fingerprinting of the gyrA/pflA genes for rapid order ing of strains by genotypic relatedness and providing additional infor mation for estimating the degree of linkage between strains was demons trated.