Identification of Trypanosoma brucei gambiense by PCR amplification of variant surface glycoprotein genes

Abstract

We have developed a sensitive and specific method to identify Trypanosoma brucei gambiense using the polymerase chain reaction (PCR) to amplify the gene encoding variant surface glycoprotein (VSG) Antat 11.17. The test was capable of distinguishing T. b. gambiense from T. b. brucei in most foci of gambian sleeping sickness and gave positive results with previously well-characterised Type I T. b. gambiense stocks from Ivory Coast, Nigeria, Cameroon, Congo, Zaire and Sudan. The test gave negative results with T. b. rhodesiense from Zambia, Kenya and Uganda, virulent or Type II T. b. gambiense from Ivory Coast and T. b. brucei stocks from East and West Africa. The test was modified for colorimetric detection in dot blot format by using nested biotinylated primers in a two-step reaction. Comparison of DNA sequences of VSG genes from T. b. gambiense and other T. brucei ssp. stocks showed a high level of homology, suggesting recent gene flow.