Detection of seven species of pathogenic leptospires by PCR using two sets of primers.
Two sets of primers derived from genomic DNA libraries of Leptospira serovars icterohaemorrhagiae (strain RGA) and bim (strain 1051) enabled the amplification by PCR of target DNA fragments from leptospiral reference strains belonging to all presently described pathogenic Leptospira species. The icterohaemorrhagiae-derived primers (G1/G2) enabled amplification of DNA from L. interrogans, L. borgpetersenii, L. weilii, L. noguchii, L. santarosai and L. meyeri, whereas the bim-derived primers (B64-I/B64-II) enabled the amplification of L. kirschneri. Southern blot and DNA sequence analysis revealed inter-species DNA polymorphism within the region spanned by primers G1 and G2 between L. interrogans and various other Leptospira species. Using a mixture of primer sets G1/G2 and B64-I/B64-II, leptospires of serovars icterohaemorrhagiae, copenhageni, hardjo, pomona, grippotyphosa and bim were detected in serum samples collected from patients during the first 10 days after the onset of illness.
Gravekamp
C
C
NH Swellengrebel Laboratory for Tropical Hygiene, Royal Tropical Institute, WHO/FAO Collaborating Centre for Reference and Research on Leptospirosis, Amsterdam, The Netherlands.
Van de Kemp
H
H
Franzen
M
M
Carrington
D
D
Schoone
G J
GJ
Van Eys
G J
GJ
Everard
C O
CO
Hartskeerl
R A
RA
Terpstra
W J
WJ
D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D015723:Gene Library; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000302:isolation & purification* / Q000472:pathogenicity; D007922:Leptospirosis / Q000175:diagnosis / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004269:DNA, Bacterial