Development of diagnostic reagents to differentiate between Mycobacterium bovis BCG vaccination and M. bovis infection in cattle

H. M. Vordermeier*, P. C. Cockle, A. Whelan, S. Rhodes, N. Palmer, D. Bakker, R. G. Hewinson

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

138 Citations (SciVal)


In Great Britain a recent independent scientific review for the government has concluded that the development of a cattle vaccine against Mycobacterium bovis holds the best long-term prospect for tuberculosis control in British herds. A sine qua non for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected with M. bovis so that test-and-slaughter-based control strategies can continue alongside vaccination. In order to assess the feasibility of developing a differential diagnostic test for a live vaccine, we chose M. bovis BCG Pasteur as a model system. Recombinant forms of antigens which are expressed in M. bovis but not, or only at low levels, in BCG Pasteur (ESAT-6, MPB64, MPB70, and MPB83) were produced. These reagents were tested either alone or in combination by using peripheral blood mononuclear cells from M. bovis-infected, BCG-vaccinated, and Mycobacterium avium-sensitized calves. All four antigens induced in vitro proliferation and gamma interferon responses only in M. bovis-infected animals. A cocktail composed of ESAT-6, MPB64, and MPB83 identified infected animals but not those vaccinated with BCG. In addition, promiscuous T-cell epitopes of ESAT- 6, MPB64, and MPB83 were formulated into a peptide cocktail. In T-cell assays with this peptide cocktail, infected animals were identified with frequencies similar to those obtained in assays with the protein cocktail, while BCG- vaccinated or M. avium-sensitized animals did not respond. In summary, our results suggest that peptide and protein cocktails can be designed to discriminate between M. bovis infection and BCG vaccination.

Original languageEnglish
Pages (from-to)675-682
Number of pages8
JournalClinical and Diagnostic Laboratory Immunology
Issue number5
Publication statusPublished - 01 Sept 1999


  • Amino Acid Sequence
  • Animals
  • Antigens, Bacterial/chemistry
  • BCG Vaccine/chemistry
  • Biomarkers
  • Cattle
  • Diagnosis, Differential
  • Interferon-gamma/immunology
  • Interleukin-2/analysis
  • Leukocytes, Mononuclear/immunology
  • Lymphocyte Activation/drug effects
  • Molecular Sequence Data
  • Mycobacterium Infections/diagnosis
  • Mycobacterium avium/immunology
  • Mycobacterium bovis/immunology
  • Peptide Fragments/chemistry
  • Recombinant Proteins/chemistry
  • Tuberculin Test
  • Tuberculosis, Bovine/diagnosis


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