Analysis of Borrelia burgdorferi sensu lato isolated from cerebrospinal fluid^Note

Abstract

Involvement of the nervous system in Lyme borreliosis may occur with or without erythema migrans and it may present with a variety of neurological symptoms. In this study we analysed phenotypic and genotypic characteristics of 40 Borrelia strains isolated from cerebrospinal fluid (CSF) of 38 Slovenian patients with different clinical manifestations of Lyme borreliosis. In seven of the patients, Borreliae were also isolated from skin lesions. Species identification and plasmid profiles were determined by pulsed-field gel electrophoresis and protein profiles by SDS-PAGE. MluI digestion profiles of Borrelia burgdorferi sensu lato DNA showed that 25 (62.5%) isolates were B. garinii, 14 (35%) B. afzelii, and one (2.5%) B. burgdorferi sensu stricto. All strains, except one, possessed a large plasmid and a varying number of smaller plasmids. Three (7.5%) isolates exhibited an unusual plasmid profile, with a large plasmid dimer or three copies of the large plasmid. In protein analyses, all strains expressed OspA protein. OspB was present significantly more often in B. afzelii than B. garinii strains (p=0.0000), while OspC was more often present in B. garinii than B. afzelii strains (p=0.0052). In the seven patients with Borreliae isolated also from the skin, the CSF and skin isolates were identical, either B. garinii (six patients) or B. afzelii (one patient). Species and plasmid heterogeneity as well as antigen diversity could play a role in the pathogenesis of the infection. When combined with our own earlier data, the results suggest species-related organotropism.

Comparison of Borrelia burgdorferi Sensu Lato Strains Isolated from Specimens Obtained Simultaneously from Two Different Sites of Infection in Individual Patients

“…Strain identification was done by pulsed-field gel electrophoresis (PFGE) as described previously (5,(33)(34)(35). Briefly, Borrelia strains were mixed in agarose, and whole blocks were lysed in lysis buffer containing lysozyme (1 mg/ml) and RNase (10 g/ml) and digested in digestion buffer containing proteinase K (0.5 mg/ml).…”

“…Skin and CSF were inoculated into modified Kelly-Pettenkofer medium immediately after they were obtained from patients (33,35). Citrated blood samples were transported to the laboratory where they were processed, and only plasma was inoculated in modified Kelly-Pettenkofer medium (34).…”

“…It has not been very unusual that an individual tick or reservoir host has harbored more then one Borrelia species (30,36,49), but data on the presence of multiple Borrelia species in an individual human patient with Lyme borreliosis have been much more limited (11,13,21,31,35). It is of interest that the majority of such mixed infections were demonstrated by PCR, while as a rule only one species appeared after culturing of the sample (18,24,25,29,47,49).…”

“…), ticks, and reservoir hosts (9,17,25,29,42,49) obtained worldwide have been analyzed by many different phenotypic methods, such as well-established protein profile and serotyping system based on the heterogeneity of OspA and OspC proteins (33)(34)(35)(42)(43)(44)(45), and several genotypic approaches (42). Analysis of borrelial chromosomal or plasmid DNA, as performed by genotypic methods, provides more precise information on the genetic relationships among Borrelia strains (2, 5, 7, 22, 33-35, 42, 47).…”

Lyme borreliosis: insights into tick- / host-borrelia relations

“…Ultimately, the development of suitable animal models for the investigation of tissue tropisms of different B. burgdorferi s.l. species will provide a more direct evidence for the correlation between Borrelia species and clinical symptoms of LB (Ružić-Sabljić et al 2001). …”