Improved serodiagnosis of erythema migrans using novel recombinant borrelial BBK32 antigens

Abstract

The performances of recombinant borrelial BBK32 proteins as antigens in the serology of erythema migrans (EM) were evaluated in an ELISA. Serum samples were obtained from 75 patients from different geographic areas where three borrelial species, Borrelia burgdorferi sensu stricto, Borrelia afzelii or Borrelia garinii, cause Lyme borreliosis. Antibodies to variant BBK32 proteins were compared with anti-flagella or with anti-IR 6 peptide antibodies. In IgG ELISA at presentation of EM, 65/75 (87 %) patients had antibodies to one or more variants of BBK32, 29/75 (39 %) had antibodies to flagella and 29/75 (39 %) had antibodies to the VlsE IR 6 peptide antigen. The immunoreactivity against variant BBK32 proteins differed in patients from different geographic regions. The present results suggest that the BBK32 proteins used in combination or in parallel may improve the laboratory diagnosis of EM.

Effects of vlsE Complementation on the Infectivity of Borrelia burgdorferi Lacking the Linear Plasmid lp28-1

“…Lyme disease patients mount a robust antibody response directed towards VlsE (29,33), and patient sera have been shown to react strongly with the IR6 invariable region of the protein (1,16,28,31,32,39,40,44). With experimentally infected mice, Triton X-114 extraction studies indicate that VlsE is present at high levels in joint and ear tissues but not in heart tissue (9), suggesting differential expression.…”

“…vlsE variation has been shown to occur within 4 days of experimental infection of mice with B. burgdorferi B31 and continues throughout the course of infection but has not been observed in vitro or in the tick vector (21, 53, 54). The conservation of vls sequences in other strains and species of Borrelia indicate that the vls locus is important for the life cycle of Lyme disease agents (23,25,49).Lyme disease patients mount a robust antibody response directed towards VlsE (29,33), and patient sera have been shown to react strongly with the IR6 invariable region of the protein (1,16,28,31,32,39,40,44). With experimentally infected mice, Triton X-114 extraction studies indicate that VlsE is present at high levels in joint and ear tissues but not in heart tissue (9), suggesting differential expression.…”

Identification of Borrelia burgdorferi Ribosomal Protein L25 by the Phage Surface Display Method and Evaluation of the Protein's Value for Serodiagnosis

“…Serological tests are widely used despite several shortcomings, such as the heterogeneity of antigen preparations and a lack of standardization causing interlaboratory variation (3,23). Furthermore, serological tests show insensitivity in the early stages of LB (1,13). It is well established that Borrelia species express different surface components depending on the temperature (21,27), pH (4,21) and cell density (30).…”

“…Therefore, assays using antigens derived from Borrelia cultures might not represent antigens which are selectively expressed in the human host. Recent studies with the recombinant proteins BBK32 and VlsE as well as two synthetic peptides have proven to be superior to assays currently used for LB serodiagnosis (1,13,24).…”

“…These clones carried inserts of 16 different sizes, which could be matched with nine genomic sequences of B. burgdorferi B31, namely, BB0182, BB0272, BB0335, BB0371, BB0713, BB0786, BBA26, BBK32, and BBL38. One sequence encoded the fibronectin binding protein BBK32, a well-established immunogenic protein (9) with high potential as a target for serodiagnosis of human LB (11,13). Another sequence identified from the B. burgdorferi and mixed libraries encoded the ribosomal protein L25.…”