Comparison of Borrelia burgdorferi sensu lato strains isolated from specimens obtained simultaneously from two different sites of infection in individual patients

Abstract

The aim of the present study was to analyze and compare Borrelia strains isolated from two different specimens obtained simultaneously from individual patients with Lyme borreliosis. Fifty such patients and 50 corresponding pairs of Borrelia isolates (100 low-propagated strains) were subjected to genotypic and phenotypic analysis, including pulsed-field gel electrophoresis for species identification and plasmid profile determination and protein profile electrophoresis for the assessment of the presence and molecular masses of separated proteins. The strains were isolated from two distinct skin lesions (12 patients), skin and blood (28 patients), skin and cerebrospinal fluid (8 patients), and blood and cerebrospinal fluid (2 patients). Out of 100 isolates, 63 were typed as B. afzelii and 37 as B. garinii. From each individual specimen only a single Borrelia species was cultured. Comparison of 50 Borrelia strain pairs isolated from two different specimens of an individual patient revealed that 12/50 (24%) patients were simultaneously infected with two different Borrelia strains; in 3/50 (6%) patients strains differed at the species level, in 4 out of the remaining 47 (9%) patients a strain difference in plasmid profile was established, while 5 out of the remaining 43 (11%) patient strain pairs differed in regard to the protein profiles of the two concurrently isolated strains. The results of the present study indicate that human patients with Lyme borreliosis may simultaneously harbor different B. burgdorferi sensu lato strains.

FIG. 1.

RFLP of Borrelia burgdorferi sensu lato strains digested with MluI restriction enzyme. MW, molecular size marker; lane 1, B. afzelii; lane 2, B. garinii; lane 3, B. burgdorferi sensu stricto strains. Isolates from patients: lanes 4 and 5, B. garinii; lanes 6 to 11, B. afzelii.

FIG. 2.

Plasmid profiles of some strain pairs. MW, molecular size marker. Isolates from patients A to D exhibit different plasmid profiles, while in patients E to G plasmid profiles of two strains (1 and 2) are congruent. In patients A to C and G B. afzelii was identified, while patients D and F were infected with B. garinii.

FIG. 3.

Protein profiles of some strain pairs. MW, molecular mass markers (106, 77, 51, 36, 28, and 21 kDa; Bio-Rad, Germany). Patients 1, 4 to 7, and 9 have different protein profiles, while patients 2, 3, and 8 exhibit identical protein profiles of both strains (1 and 2) of the corresponding pair.