Peptide-Based OspC Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Lyme Borreliosis

Abstract

Sera from 210 patients with Lyme borreliosis (LB) were studied by an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (pepC10) comprising the C-terminal 10-amino-acid residues of OspC of Borrelia burgdorferi. We found that 36.3 and 45.0% of the serum samples from patients with erythema migrans (EM) and neuroborreliosis (NB), respectively, displayed immunoglobulin M (IgM) anti-pepC10 reactivities, while these samples rarely (≤8%) displayed IgG antibody reactivities. Sera from patients with acrodermatitis chronica atrophicans did not contain anti-pepC10 antibodies. The diagnostic performance of this newly developed peptide ELISA was compared with those of an ELISA based on the full-length recombinant OspC protein (rOspC) and a commercially available ELISA based on theB. burgdorferi flagellum (Fla). The sensitivity of the IgM pepC10 ELISA was slightly lower (P < 0.04) than that of the rOspC ELISA for EM patients (36.3 versus 43.8%), while there was no difference for NB patients (45.0 versus 48.0%). However, the optical density values obtained by the pepC10 ELISA were generally higher than those obtained by the rOspC ELISA, leading to a significantly better quantitative discrimination between seropositive patients with NB and controls (P < 0.008). The specificity of the pepC10 ELISA was similar to those of the rOspC ELISA and the Fla ELISA for relevant controls including patients with syphilis and mononucleosis. Although the overall diagnostic sensitivity of the Fla ELISA was superior, 8.8 and 12.0% of the EM and NB patients, respectively, were antibody positive only by the pepC10 ELISA. Thus, use of a diagnostic test for LB based on the detection of IgM antibodies to pepC10 and Fla has increased sensitivity for the diagnosis of early LB.

Serodiagnosis of Lyme Disease by Kinetic Enzyme‐Linked Immunosorbent Assay Using Recombinant VlsE1 or Peptide Antigens ofBorrelia burgdorferiCompared with 2‐Tiered Testing Using Whole‐Cell Lysates

“…pepC10 peptide synthesis. pepC10 (PVVAESPKKP) [18] was synthesized by the Biotechnology Core Facility (Scientific Resources Program, National Center for Infectious Diseases, CDC, Atlanta) using Fmoc chemistry. The amino terminus was derivatized with sulfo-NHS-LC-biotin (Pierce), which introduced a 6-carbon spacer between the peptide and biotin.…”

“…Recently, substantial progress has been made in developing new serologic tests based on recombinant antigens and synthetic peptides [15][16][17][18][19][20][21][22][23][24][25]. We evaluated 3 antigens in a kinetic ELISA (kELISA) format [26][27][28][29] and compared these tests with the results of standardized 2-tiered testing.…”

“…Finally, we assessed whether a kELISA based on a conserved peptide of outer-surface protein C (OspC) would be a beneficial supplement to either the rVlsE1 or the C6 test. Detection of IgM antibodies to a 10-aa peptide found at the C terminus of most OspC proteins (pepC10) has been demonstrated to be helpful in the diagnosis of acute Lyme disease and neuroborreliosis in Europe [18,19].…”

A host-restricted viral vector for antigen-specific immunization against Lyme disease pathogen

“…Two borrelial gene products, the outer membrane antigens BmpA and OspC, were chosen as candidate immunogens for the study. These proteins are immunogenic in diverse species including humans [34,38] and are known to confer strain-specific immunity against B. burgdorferi infection in mice [40]. Both antigens are lipoproteins that contain type II signal peptides absent in the mature proteins.…”

“…BmpA and OspC are examples of outer membrane antigens that are produced by the pathogens during infection [34][35][36][37]. Robust antibody responses against both antigens are detectable in infected hosts including humans that are useful for serodiagnosis of Lyme disease [38,39]. In addition, while OspC immunization results in generation of strain-specific immunity [40][41][42], BmpA immunization interferes with B. burgdorferi persistence in a tissue-specific manner, only in the joints [36].…”