Serodiagnosis of bovine trypanosomosis based on HSP70/BiP inhibition ELISA.
Animal trypanosomosis is a serious constraint to livestock productivity in tropical and sub-tropical countries. The pathogenic trypanosomes in bovidae are Trypanosoma congolense, T. vivax, T. brucei and T. evansi. Current serological tests to detect trypanosome infections are based on the use of whole trypanosome lysates; their potential is limited by antigen instability, lack of reproducibility and lack of test specificity due to the antibody's long persistence after treatment. The development of new tests based on recombinant technology that could be standardized and applied on a large scale at low cost would be very helpful. The major invariant antigen recognized by T. congolense infected cattle belongs to the heat shock protein (HSP) 70 family and is closely related to mammalian Immunoglobulin Binding Protein (BiP). To improve the initial ELISA based on a recombinant fragment of HSP70/BiP, we developed an inhibition ELISA using an anti-BiP monoclonal antibody and a full-length fusion protein expressed in E. coli. Here we report on the development of the test and provide an initial assessment of its performance using sets of sera from experimental infections and from naturally infected cattle maintained in tsetse infested areas of Africa. The HSP70/BIP-based inhibition ELISA shows a good sensitivity in cattle experimentally infected with T. congolense, with an improved sensitivity in secondary infections. One major advantage, particularly for its further application in national laboratories, is that one single set of reagents and one single procedure are sufficient to apply on different mammalian host species infected with different trypanosome species.
Bossard
Geraldine
G
UMR17 Trypanosomes, IRD-CIRAD, Campus International de Baillarguet, F-34398 Montpellier, France. bossard@cirad.fr
Boulange
Alain
A
Holzmuller
Philippe
P
Thévenon
Sophie
S
Patrel
Delphine
D
Authie
Edith
E
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D015800:Protozoan Proteins / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000662:veterinary*; D014348:Trypanosoma congolense / Q000276:immunology; D014353:Trypanosomiasis, African / Q000097:blood / Q000175:diagnosis / Q000662:veterinary*; D014354:Trypanosomiasis, Bovine / Q000097:blood / Q000175:diagnosis*
D000911:Antibodies, Monoclonal; D018840:HSP70 Heat-Shock Proteins; D015800:Protozoan Proteins