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Clinical and Diagnostic Laboratory Immunology, September 1999, p. 675-682, Vol. 6, No. 5
1071-412X/99/$04.00+0

Development of Diagnostic Reagents To Differentiate between Mycobacterium bovis BCG Vaccination and M. bovis Infection in Cattle

H. M. Vordermeier,1,* P. C. Cockle,1 A. Whelan,1 S. Rhodes,1 N. Palmer,2 D. Bakker,3 and R. G. Hewinson1

TB Research Group,1 and TB Diagnostic Unit,2 Bacteriology Department, Veterinary Laboratories Agency---Weybridge, New Haw, Addlestone, KT15 3NB, United Kingdom, and Department of Small Ruminants Health, Animal Health Service, Boxtel, The Netherlands3

Received 13 January 1999/Accepted 20 May 1999

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.

* Corresponding author. Mailing address: TB Research Group, Bacteriology Department, Veterinary Laboratories Agency---Weybridge, New Haw, Addlestone, KT15 3NB, United Kingdom. Phone: 44 1932 357 884. Fax: 44 1932 357 684. E-mail: mvordermeier.vla{at}gtnet.gov.uk.

Clinical and Diagnostic Laboratory Immunology, September 1999, p. 675-682, Vol. 6, No. 5
1071-412X/99/$04.00+0


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