Large differences between test strategies for the detection of anti-Borrelia antibodies are revealed by comparing eight ELISAs and five immunoblots

Eur J Clin Microbiol Infect Dis. 2011 Aug;30(8):1027-32. doi: 10.1007/s10096-011-1157-6. Epub 2011 Jan 27.

Abstract

We investigated the influence of assay choice on the results in a two-tier testing algorithm for the detection of anti-Borrelia antibodies. Eighty-nine serum samples from clinically well-defined patients were tested in eight different enzyme-linked immunosorbent assay (ELISA) systems based on whole-cell antigens, whole-cell antigens supplemented with VlsE and assays using exclusively recombinant proteins. A subset of samples was tested in five immunoblots: one whole-cell blot, one whole-cell blot supplemented with VlsE and three recombinant blots. The number of IgM- and/or IgG-positive ELISA results in the group of patients suspected of Borrelia infection ranged from 34 to 59%. The percentage of positives in cross-reactivity controls ranged from 0 to 38%. Comparison of immunoblots yielded large differences in inter-test agreement and showed, at best, a moderate agreement between tests. Remarkably, some immunoblots gave positive results in samples that had been tested negative by all eight ELISAs. The percentage of positive blots following a positive ELISA result depended heavily on the choice of ELISA-immunoblot combination. We conclude that the assays used to detect anti-Borrelia antibodies have widely divergent sensitivity and specificity. The choice of ELISA-immunoblot combination severely influences the number of positive results, making the exchange of test results between laboratories with different methodologies hazardous.

Publication types

  • Evaluation Study

MeSH terms

  • Antibodies, Bacterial / blood*
  • Antigens, Bacterial / immunology
  • Borrelia / immunology*
  • Clinical Laboratory Techniques / methods*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Humans
  • Immunoblotting / methods*
  • Immunoglobulin G / blood
  • Immunoglobulin M / blood
  • Lyme Disease / diagnosis*
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity

Substances

  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Immunoglobulin G
  • Immunoglobulin M
  • Recombinant Proteins