Genotypic and phenotypic characterisation of Borrelia burgdorferi sensu lato strains isolated from human blood.
Lyme borreliosis often presents initially with erythema migrans. Borreliae may disseminate from the primary skin lesion, and different organs and systems could be affected. Borrelia strains were isolated from blood of 70 patients with Lyme borreliosis, including 10 patients from whom borreliae were also isolated from skin. The aim of the present study was to characterise the isolates with regard to their phenotypic and genotypic characteristics. Borreliae were cultivated in MKP medium. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis (PFGE) and protein profiles by SDS-PAGE. Digestion of Borrelia burgdorferi sensu lato DNA showed 63 (90%) B. afzelii Mla1 and 7 (10%) B. garinii Mlg2. No B. burgdorferi sensu stricto were isolated. Borreliae were isolated from both skin and blood of 10 patients, nine pairs of isolates were identical: seven B. afzelii and two B. garinii. B. afzelii was isolated from the skin and B. garinii from blood of the tenth patient. All but one isolate possessed at least one large plasmid and varying numbers of smaller plasmids. Eight (11.4%) of 70 isolates possessed an unusual plasmid profile (2 of 63 B. afzelii and 6 of 7 B. garinii). Borreliae differed in their protein profiles. OspA and OspB proteins were expressed by all B. afzelii isolates; 85.7% of B. garinii isolates expressed OspA and 71.4% expressed OspB. OspC was expressed by 65% of B. afzelii isolates and all B. garinii isolates. The ratios of B. afzelii and B. garinii isolated from blood and skin were similar. These results do not support the hypothesis that B. garinii has a higher propensity for haematogenous dissemination than B. afzelii. Antigen diversity as well as species and plasmid heterogeneity could play a role in the pathogenesis of the infection, suggesting distinctive strain organotropism.
Ružić-Sabljić
E
E
Arnež
M
M
Institute of Microbiology and Immunology, Medical Faculty Ljubljana, Zaloška 4 and *Department of Infectious Diseases, University Medical Centre, Japljeva 2, 1000 Ljubljana, Slovenia.
Lotrič-Furlan
S
S
Institute of Microbiology and Immunology, Medical Faculty Ljubljana, Zaloška 4 and *Department of Infectious Diseases, University Medical Centre, Japljeva 2, 1000 Ljubljana, Slovenia.
Maraspin
V
V
Institute of Microbiology and Immunology, Medical Faculty Ljubljana, Zaloška 4 and *Department of Infectious Diseases, University Medical Centre, Japljeva 2, 1000 Ljubljana, Slovenia.
Cimperman
J
J
Institute of Microbiology and Immunology, Medical Faculty Ljubljana, Zaloška 4 and *Department of Infectious Diseases, University Medical Centre, Japljeva 2, 1000 Ljubljana, Slovenia.
Strle
F
F
Institute of Microbiology and Immunology, Medical Faculty Ljubljana, Zaloška 4 and *Department of Infectious Diseases, University Medical Centre, Japljeva 2, 1000 Ljubljana, Slovenia.
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics; D015748:Borrelia burgdorferi Group / Q000145:classification* / Q000235:genetics / Q000502:physiology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D004890:Erythema / Q000097:blood / Q000382:microbiology; D006801:Humans; D008193:Lyme Disease / Q000097:blood / Q000453:epidemiology / Q000382:microbiology*; D017192:Skin Diseases, Bacterial / Q000097:blood / Q000453:epidemiology / Q000382:microbiology; D017524:Slovenia / Q000453:epidemiology
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial