BACKGROUND: The World Health Organization targeted Trypanosoma brucei gambiense human African trypanosomiasis (gHAT) for elimination as a public health problem and for elimination of transmission. To measure gHAT elimination success with prevalences close to zero, highly specific diagnostics are necessary. Such a test exists in the form of an antibody-mediated complement lysis test, the trypanolysis test, but biosafety issues and technological requirements prevent its large-scale use. We developed an inhibition ELISA with high specificity and sensitivity that is applicable in regional laboratories in gHAT endemic countries.
METHODS: The T. b. gambiense inhibition ELISA (g-iELISA) is based on the principle that binding of monoclonal antibodies to specific epitopes of T. b. gambiense surface glycoproteins can be inhibited by circulating antibodies of gHAT patients directed against the same epitopes. Using trypanolysis as reference test, the diagnostic accuracy of the g-iELISA was evaluated on plasma samples from 739 gHAT patients and 619 endemic controls and on dried blood spots prepared with plasma of 95 gHAT and 37 endemic controls.
RESULTS: Overall sensitivity and specificity on plasma were respectively 98.0% (95% CI 96.7 - 98.9) and 99.5% (95% CI 98.6-99.9). With dried blood spots, sensitivity was 92.6% (95% CI 85.4 - 97.0), and specificity was 100% (95% CI 90.5 - 100.0). The g-iELISA is stable for at least 8 months when stored at 2-8°C.
CONCLUSION: The g-iELISA might largely replace trypanolysis for monitoring gHAT elimination and for post-elimination surveillance. The g-iELISA kit is available for evaluation in reference laboratories in endemic countries.