Abstract
Borrelia burgdorferi is the causative agent of Lyme borreliosis, which is the most common tick-borne human disease in Europe and North America. Currently, the diagnosis of Lyme borreliosis is based on serological tests allowing indirect detection of anti-Borrelia antibodies produced by patients. Their main drawback is a lack of sensitivity in the early phase of disease and an incapacity to prove an active infection. Direct diagnostic tests are clearly needed. The objectives of this study were to produce tools allowing sensitive detection of potential circulating Borrelia antigens and to evaluate them in a mouse model. We focused on two potential early bacterial makers, the highly variable OspC protein and the conserved protein FlaB. High-affinity monoclonal antibodies were produced and used to establish various immunoassays and western blot detection. A very good limit of detection for OspC as low as 17 pg/mL of sample was achieved with SPIE-IA. In infected mice, we were able to measure OspC in plasma with a mean value of 10 ng/mL at 7 days post-inoculation. This result suggests that OspC could be a good blood marker for diagnosis of Lyme borreliosis and that the tools developed during this study could be very useful.
Highlights
Borrelia burgdorferi is the causative agent of Lyme borreliosis, which is the most common tick-borne human disease in Europe and North America
We focused on two Borrelia proteins, flagellin B (FlaB), a major component of the periplasmic flagellar filament crucial for bacterial mobility, and outer surface protein C (OspC), a lipoprotein needed for the establishment of early localized infection
The first measures antibody binding with biotinylated recombinant FlaB and the second measures binding with whole-cell Borrelia burgdorferi extract coated on a solid phase
Summary
Borrelia burgdorferi is the causative agent of Lyme borreliosis, which is the most common tick-borne human disease in Europe and North America. The diagnosis of Lyme borreliosis is based on serological tests allowing indirect detection of anti-Borrelia antibodies produced by patients. Their main drawback is a lack of sensitivity in the early phase of disease and an incapacity to prove an active infection. The recommended approach for laboratory diagnosis of Lyme borreliosis is the two-tiered testing strategy, which includes an indirect enzyme-linked immunosorbent assay (ELISA) followed by a more specific western blot in order to rule out false positives[11] These tests do not prove the existence of an active infection and are not recommended in the early localized phase of the disease. A new method based on the concentration of outer surface protein A (OspA) in a large volume of urine followed by detection with western blot was reported to be efficient in the early detection of active infection[20], but robust validation is still needed
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