Diagnosis of human trypanosomiasis, due to Trypanosoma brucei gambiense in central Africa, by the polymerase chain reaction

Abstract

During a mass screening of sleeping sickness conducted in 1998 and 1999, and involving 27 932 persons in Cameroon and the Central African Republic, we tested the polymerase chain reaction (PCR) on whole blood for the diagnosis of human African trypanosomiasis due to Trypanosoma brucei gambiense. The 1858 samples obtained were from 4 groups: 155 infected patients, 1432 serological suspects detected by the card agglutination test for trypanosomiasis (CATT), 222 negative controls living in the prospected area (negative with the CATT and parasitological methods), and 49 negative controls (CATT and parasitological methods) and unexposed to the disease (Europeans). The technique of DNA extraction used made it possible to preserve the blood samples in the field. The primers used were specific for T. brucei s.l. Only 1 patient was PCR negative, and 3 of the negative controls, exposed to the disease, were PCR positive. Among the 1432 serological suspects, only 50 were PCR positive. During the 6-month follow-up after the surveys, the 3 negative controls, who were initially positive by PCR, were found to be negative. These initial positive PCR results are unlikely to have been due to a cross-reaction with T. brucei brucei, which is non-pathogenic for man, but are more likely to have resulted from a mislabelling of sample tubes. All control individuals, exposed or not to the disease, were negative by PCR. The PCR-negative patient was possibly a registration error. Among 50 PCR positive serological suspects, 39 of them were re-examined. Five were found to be positive by the kit for in-vitro isolation of trypanosomes, representing an increase in patients of almost 13%. At the end of the study, 160 patients were diagnosed, and the PCR was positive for 159 of them (99·4%). Moreover, the PCR made it possible to reduce the number of suspects to be re-examined (50 instead of 1432; a reduction of 96·5%).