Use of Synthetic Peptides Derived from the Antigens ESAT-6 and CFP-10 for Differential Diagnosis of Bovine Tuberculosis in Cattle

doi:10.1128/CDLI.8.3.571-578.2001

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Clinical and Diagnostic Laboratory Immunology, May 2001, p. 571-578, Vol. 8, No. 3
1071-412X/01/$04.00+0 DOI: 10.1128/CDLI.8.3.571-578.2001

Use of Synthetic Peptides Derived from the Antigens ESAT-6 and CFP-10 for Differential Diagnosis of Bovine Tuberculosis in Cattle

H. M. Vordermeier,¹^,^* A. Whelan,¹ P. J. Cockle,¹ L. Farrant,² N. Palmer,³ and R. G. Hewinson¹

TB Research Group¹ and TB Diagnostic Section,³ Department of Bacterial Diseases, Veterinary Laboratories Agency-Weybridge, New Haw, Addlestone KT15 3NB, and Ministry of Agriculture, Fisheries and Food, State Veterinary Service, Exeter EX5 1DY,² United Kingdom

Received 24 July 2000/Returned for modification 6 December 2000/Accepted 9 February 2001

In Great Britain an independent scientific review for the government has concluded that the development of a cattle vaccineagainst Mycobacterium bovis infection holds the best long-termprospect for tuberculosis control in British herds. A preconditionfor vaccination is the development of a complementary diagnostictest to differentiate between vaccinated animals and those infectedwith M. bovis so that testing and slaughter-based control strategiescan continue alongside vaccination. To date bacillus Calmette-Guérin(BCG), an attenuated strain of M. bovis, is the only availablevaccine for the prevention of tuberculosis. However, tests basedon tuberculin purified protein derivative cannot distinguish betweenM. bovis infection and BCG vaccination. Therefore, specific antigensexpressed by M. bovis but absent from BCG constitute prime candidatesfor differential diagnostic reagents. Recently, two such antigens,ESAT-6 and CFP-10, have been reported to be promising candidatesas diagnostic reagents for the detection of M. bovis infectionin cattle. Here we report the identification of promiscuous peptidesof CFP-10 that were recognized by M. bovis-infected cattle. Fiveof these peptides were formulated into a peptide cocktail togetherwith five peptides derived from ESAT-6. Using this peptide cocktailin T-cell assays, M. bovis-infected animals were detected, whileBCG-vaccinated or Mycobacterium avium-sensitized animals did notrespond. The sensitivity of the peptide cocktail as an antigenin a whole-blood gamma interferon assay was determined using naturallyinfected field reactor cattle, and the specificity was determinedusing blood from BCG-vaccinated and noninfected, nonvaccinatedanimals. The sensitivity of the assay in cattle with confirmedtuberculosis was found to be 77.9%, with a specificity of 100%in BCG-vaccinated or nonvaccinated animals. This compares favorablywith the specificity of tuberculin when tested in noninfectedor vaccinated animals. In summary, our results demonstrate thatthis peptide cocktail can discriminate between M. bovis infectionand BCG vaccination with a high degree of sensitivity andspecificity.

^* Corresponding author. Mailing address: TB Research Group, Department of Bacterial Diseases, Veterinary Laboratories Agency-Weybridge,New Haw, Addlestone KT15 3NB, United Kingdom. Phone: 44 1932 357884. Fax: 44 1932 357 684. E-mail: mvordermeier.vla{at}gtnet.gov.uk.

Clinical and Diagnostic Laboratory Immunology, May 2001, p. 571-578, Vol. 8, No. 3
1071-412X/01/$04.00+0 DOI: 10.1128/CDLI.8.3.571-578.2001

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