Development and application of the technique for identification of Borrelia miyamotoi surface antigens

Abstract

Introduction. Borrelia miyamotoi is a pathogen causing erythema-free ixodid tick-borne borreliosis (ITBB), a disease widespread in Russia. The genome of B. miyamotoi contains genes of multiple variable major proteins (Vmps). Vmps fall into two families — Vsps and Vlps (with subfamilies δ, γ, α and β). At a particular time, a single B. miyamotoi expresses only one variant of Vmp gene.The purpose of the work is to develop a technique for identification of the Vmp present at the expression site.Materials and methods. The technique is designed in the format of a real-time multiplex PCR. It was tested by using B. miyamotoi DNA samples extracted from blood collected from 172 ITBB patients and 109 ticks. The samples were collected in 14 regions of Russia.Results. The new technique made it possible to identify the expressed Vmp in 82% of the examined samples, thus having demonstrated its efficiency. Negative results were much less often observed with samples from patients than with samples from ticks. At the same time, the percentage of samples with one type of Vmp is identical for clinical samples and ticks, while the percentage of samples containing concurrently two types of Vmps is significantly higher among samples from patients with the most frequent occurrence of the Vlp-δ and Vsp combination.Discussion. The frequent occurrence of the combination of two Vmp types in the blood samples can indicate the concurrent presence of several subpopulations of B. miyamotoi in ITBB patients. A new antigenic Vmp variant is synthesized after protective antibodies have been produced for the major protein of the strain transmitted by a tick. This phenomenon known as immune evasion allows the pathogen to persist within a host.Conclusion. The developed technique of real-time multiplex PCR allows to simultaneous detect of several antigenic variants of the variable basic surface proteins of B. miyamotoi. The study of the antigenic spectrum of B. miyamotoi strains in comparison with the characteristics of conserved regions of the genome by the method of multilocus sequencing will clarify the stages of evolution and distribution of B. miyamotoi sensu lato.