Detection of Trypanosoma brucei gambiense and T. b. rhodesiense in Glossina fuscipes fuscipes (Diptera: Glossinidae ) and Stomoxys flies using the polymerase chain reaction (PCR) technique in southern Sudan

Abstract

Ethanol-fixed entire bodies of the tsetse fly, Glossina fuscipes fuscipes , and unidentified stable flies, Stomoxys spp ., collected from near Juba town, southern Sudan, were  tested for Trypanozoon trypanosomes infections using polymerase chain reaction (PCR) technique for the first time in Sudan. The crude target DNA sequences were extracted by incubation of entire flies in Nonindet PCR template buffer containing proteinase-K. The DNA amplification sets of conditions were adjusted for each pair of primers employed. The oligonucleotide primers used included TBR 1-2 , SRA A-E , SRA B537-B538 and TgsGP FOR-REV . The results showed that 74.4% of G. f. fuscipes and 39.36% of Stomoxys spp . were infected with Trypanozoon trypanosomes . Out of the 117 examined G. f. fuscipes , 46.2, 24.8, 35.04, 17.09 and 10.26% were due to T. b. gambiense (TgsGP FOR-REV ), T. b. rhodesiense (SRA A-E ), T. b. rhodesiense (SRA 3537-3538 ), mixed infection with T. b. gambiense and T. b. rhodesiense and T. b. brucei, respectively. However, infections in Stomoxys spp. of 2.13 and 37.2% were due to T. b. rhodesiense and T. b. brucei , respectively. Key words : Glossina fuscipes fuscipes, T. b. gambiense, T. b. Rhodesiense, vectoral capacity, infection rate, PCR technology.