Recombinant Assay for Serodiagnosis of Lyme Disease Regardless of OspA Vaccination Status

Abstract

All current seroassays using cultured Borrelia burgdorferi as their antigen source have been rendered obsolete by the recombinant OspA Lyme disease vaccine. OspA is the major outer surface protein expressed in cultured B. burgdorferi, and any seroassay that uses whole organisms as its antigen source cannot differentiate between subjects who received the vaccine and those who were naturally infected. We developed a new sensitive and specific enzyme-linked immunosorbent assay (ELISA) utilizing recombinant chimeric borrelia proteins devoid of OspA (rNon-OspA) that can be used to detect antibodies to diagnostically important B. burgdorferi antigens in both OspA-vaccinated and nonvaccinated individuals. We tested sera from patients with Lyme disease and with conditions associated with false-positive serologies, OspA-vaccinated individuals, and healthy high-risk workers from an area of endemicity and normal sera from individuals from areas of nonendemicity. The rNon-OspA test was compared with two commercially available whole-cell immunoassays. The rNon-OspA assay is as sensitive and specific as the whole-cell assay (P > 0.05) for detection of anti-B. burgdorferi antibodies. However, the rNon-OspA assay can differentiate between populations comprised of naturally infected and OspA-vaccinated individuals (P < 0.05). Our data demonstrate that this new sensitive rNon-OspA ELISA can be used for the laboratory detection of B. burgdorferi antibodies regardless of vaccination status and could replace existing serologic assays for Lyme disease.

OspE-Related, OspF-Related, and Elp Lipoproteins Are Immunogenic in Baboons Experimentally Infected withBorrelia burgdorferiand in Human Lyme Disease Patients

“…Furthermore, the administration of the OspA-based Lyme disease vaccine has complicated the application of ELISAs based upon WCL to identify B. burgdorferi infection in vaccinated individuals (1,26,78). Recent attempts have been made to identify single antigens or a combination of antigens that result in sensitivity similar to that generated against B. burgdorferi WCL but with an increase in the specificity of reactive antibodies (15,27,30,31,40,(42)(43)(44)58). In this study, a serological analysis of reactivity to the OspE-related, OspF-related, and Elp proteins was performed on patients with early and late Lyme disease, arbitrarily defined as less than 12 weeks p.i.…”

Decorin Binding Proteins A and B in the Serodiagnosis of Lyme Disease in North America

“…The region corresponding to DbpA(6 -30) contained three overlapping peptides in the peptide library (Fig. 1); however, the peptide region DbpA (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25) was not properly synthesized due to the chemical properties of that particular amino acid sequence. As a result, 3 peptides were generated from that region for further analysis, DbpA (6 -20), DbpA (16 -30), and DbpA(6 -30), which contains the interceding region that was not produced independently; in addition, a single peptide was generated from the sequence DbpA(76 -90) (Fig.…”

“…The inclusion of the second-tier Western blot improved specificity, but as it is a less sensitive assay than the first-tier ELISA, it ultimately reduced the sensitivity of the whole two-tier paradigm. While two-tier testing has been useful, it lacks sufficient sensitivity at the time many patients with early Lyme disease usually seek initial medical care, failing to detect diagnostic levels of antibodies approximately 50% of the time (6,(8)(9)(10)(11)(12)(13). If the specificity of the first-tier assays were improved, second-tier assays could be eliminated, increasing the sensitivity of serodiagnostics.…”

Improving the Specificity of Recombinant Immunoassays for Lyme Disease

“…However, the use of recombinant FlaB and OspC, in addition to the above antigens, may have been prematurely rejected by the authors. The Hefty paper, similar to that of Gomes-Solecki et al (2), considers a serologic response to any of a group of recombinant antigens indicative of B. burgdorferi infection. This method of data analysis attempts to maximize sensitivity but fails to maximally utilize the improved specificity available through recombinant techniques.…”