DETECTION OF 7 SPECIES OF PATHOGENIC LEPTOSPIRES BY PCR USING 2 SETS OF PRIMERS

Two sets of primers derived from genomic DNA libraries of Leptospira s erovars icterohaemorrhagiae (strain RGA) and bim (strain 1051)enabled the amplification by PCR of target DNA fragments from leptospiral refe rence strains belonging to all presently described pathogenic Leptospi ra species. The icterohaemorrhagiae-derived primers (G1/G2) enabled am plification of DNA from L. interrogans, L. borgpetersenii, L. weilii, L. noguchii, L. santarosai and L. meyeri, whereas the bim-derived prim ers (B64-I/B64-II) enabled the amplification of L. kirschneri. Souther n blot and DNA sequence analysis revealed inter-species DNA polymorphi sm within the region spanned by primers G1 and G2 between L. interroga ns and various other Leptospira species. Using a mixture of primer set s G1/G2 and B64-I/B64-II, leptospires of serovars icterohaemorrhagiae, copenhageni, hardjo, pomona, grippotyphosa and bim were detected in s erum samples collected from patients during the first 10 days after th e onset of illness.