Primer reactivities of 25
Borrelia burgdorferi sensu lato isolates from the ticks,
Ixodes persulcatus and
I. ovatus, in Japan and 10 isolates in Europe and North America were investigated. The methods used in this study were the polymerase chain reaction (PCR) on the flagellin structural gene (
fla), the outer surface protein A gene (
osp A) and the outer surface protein B gene (
osp B), and the restriction fragment length polymorphism (RFLP) analysis of PCR products from
osp A and
osp B. The flagellin PCR primer set reacted with all the
Borrelia strains tested. Four genospecies,
B. burgdorferi sensu stricto,
B. garinii, B. afzelii and
B. japonica, were differentiated by PCR using
osp A and
osp B primers combined with RFLP analysis. Some Japanese isolates from
I. persulcatus were identified as
B. garinii or
B. afzelii. The other isolates from
I. persulcatus did not fit in any of the 4 genospecies.
These results suggested that Japanese isolates from
I. persulcatus are highly heterogeneous in their
osp A and
osp B structures. Furthermore, PCR primers targeting
fla are applicable to the gene diagnosis for Lyme disease in Japan, and
osp A and
osp B primers can be used to classify
B. burgdorferi sensu lato isolates into genospecies by PCR and RFLP analyses.
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