Design, construction and evaluation of multi-epitope antigens for diagnosis of Lyme disease

Ann Agric Environ Med. 2017 Dec 23;24(4):696-701. doi: 10.26444/aaem/80699. Epub 2017 Dec 18.

Abstract

Introduction: Introduction and objective. Lyme disease (LD) is the most common vector-borne disease in the temperate zone of the Northern Hemisphere. Diagnosis of LD is mainly based on clinical symptoms supported with serology (detection of anti-Borrelia antibodies) and is often misdiagnosed in areas of endemicity.

Material and methods: In this study, the chimeric proteins (A/C-2, A/C-4 and A/C-7.1) consisting of B-cell epitopes of outer surface proteins OspA and OspC from Borrelia genospecies prevalent in Eastern Slovakia, were designed, over-expressed in E. coli, and used to detect specific anti-Borrelia antibodies in serologically characterized sera from patients with Lyme-like symptoms to evaluate their diagnostic potential.

Results: Results showed that chimeras vary in their immuno-reactivity when tested with human sera. Compared with the results obtained from a two-tier test, the application of recombinant multi-epitope chimeric proteins as diagnosis antigens, produced fair agreement in the case of A/C-2 (0.20<κ<0.40) and good agreement (0.60<κ<0.80) when A/C-7.1 was used as capture antigen. Chimera A/C-4 were excluded from further study due to loss of reactivity with OspA-specific antibodies.

Conclusions: The combination of specific B-cell epitopes from OspA and OspC proteins may improve the diagnostic accuracy of serologic assays, but further studies are required to address this hypothesis.

Keywords: ELISA; Lyme disease; Slovakia; chimera; epitope.

Publication types

  • Evaluation Study

MeSH terms

  • Antibodies, Bacterial / analysis
  • Antibodies, Bacterial / immunology
  • Antigens, Surface / analysis
  • Antigens, Surface / genetics
  • Antigens, Surface / immunology
  • Bacterial Outer Membrane Proteins / analysis
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Vaccines / analysis
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology
  • Borrelia burgdorferi / genetics
  • Borrelia burgdorferi / immunology
  • Borrelia burgdorferi / isolation & purification*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes, B-Lymphocyte / analysis*
  • Epitopes, B-Lymphocyte / immunology
  • Humans
  • Lipoproteins / analysis
  • Lipoproteins / genetics
  • Lipoproteins / immunology
  • Lyme Disease / diagnosis*
  • Lyme Disease / immunology
  • Lyme Disease / microbiology

Substances

  • Antibodies, Bacterial
  • Antigens, Surface
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Epitopes, B-Lymphocyte
  • Lipoproteins
  • OspA protein