title: Genetic diversity of Borrelia burgdorferi in lyme disease patients as determined by culture versus direct PCR with clinical specimens.
abstract: Two hundred seventeen isolates of Borrelia burgdorferi originally cultured from skin biopsy samples or blood of early Lyme disease patients were genetically characterized by PCR-restriction fragment length polymorphism (RFLP) typing of the 16S-23S ribosomal DNA intergenic spacer. Three major RFLP types were observed. Of the cultured isolates, 63 of 217 (29.0%) were type 1, 85 of 217 (39.2%) were type 2, and 58 of 217 (26.7%) were type 3; mixtures of two RFLP types were obtained in 6.0% (13 of 217) of the cultures. Comparison of typing of B. burgdorferi performed directly on 51 patient skin specimens with typing of cultures originally isolated from the same tissue revealed that a much larger proportion of direct tissue samples had mixtures of RFLP types (43.1% by direct typing versus 5.9% by culture [P < 0.001). In addition, identical RFLP types were observed in only 35.5% (11 of 31) of the paired samples. RFLP type 3 organisms were recovered from blood at a significantly lower rate than were either type 1 or type 2 strains. These studies demonstrate that the genetic diversity of B. burgdorferi patient isolates as determined by cultivation differs from that assessed by PCR performed directly on patient tissue.
authors.affiliation: Departments of Biochemistry and Molecular Biology, New York Medical College, Valhalla, New York 10595, USA.
authors.forename: D
authors.initials: D
authors.lastname: Liveris
authors.forename: S
authors.initials: S
authors.lastname: Varde
authors.forename: R
authors.initials: R
authors.lastname: Iyer
authors.forename: S
authors.initials: S
authors.lastname: Koenig
authors.forename: S
authors.initials: S
authors.lastname: Bittker
authors.forename: D
authors.initials: D
authors.lastname: Cooper
authors.forename: D
authors.initials: D
authors.lastname: McKenna
authors.forename: J
authors.initials: J
authors.lastname: Nowakowski
authors.forename: R B
authors.initials: RB
authors.lastname: Nadelman
authors.forename: G P
authors.initials: GP
authors.lastname: Wormser
authors.forename: I
authors.initials: I
authors.lastname: Schwartz
mesh_terms: D001431:Bacteriological Techniques; D001706:Biopsy; D015748:Borrelia burgdorferi Group / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000235:genetics; D014644:Genetic Variation; D006801:Humans; D008193:Lyme Disease / Q000097:blood / Q000382:microbiology* / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics; D012867:Skin / Q000382:microbiology* / Q000473:pathology