DEVELOPMENT OF A MULTIPLEX PCR GENE FINGERPRINTING METHOD USING GYRA AND PFLA POLYMORPHISMS TO IDENTIFY GENOTYPIC RELATEDNESS WITHIN CAMPYLOBACTER-JEJUNI SPECIES
C. Ragimbeau et al., DEVELOPMENT OF A MULTIPLEX PCR GENE FINGERPRINTING METHOD USING GYRA AND PFLA POLYMORPHISMS TO IDENTIFY GENOTYPIC RELATEDNESS WITHIN CAMPYLOBACTER-JEJUNI SPECIES, Journal of applied microbiology, 85(5), 1998, pp. 829-838
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.