Detection and identification of Trypanosoma of African livestock through asingle PCR based on internal transcribed spacer 1 of rDNA

Primers hybridising with the rDNA cistron have previously been evaluated fo r PCR diagnosis specific for kinetoplastids, and shown to detect and differ entiate the Trypanosoma brucei complex and Trypanosoma cruzi. Kin1 and Kin2 primers, amplifying internal transcribed spacer 1, were subsequently evalu ated for the diagnosis of African livestock trypanosomosis. Based on the si ze of the PCR products obtained, Kin primers allowed detection and identifi cation of three Trypanosoma congolense types (savannah, forest and Kenya Co ast), with distinction among themselves and from the subgenus Trypanozoon ( T. brucei spp., Trypanosoma evansi and Trypanosoma equiperdum), Trypanosoma vivax, Trypanosoma simiae and Trypanosoma theileri. These primers were sho wn to be suitable for the sensitive and type-specific diagnosis of African livestock trypanosome isolates through a single PCR even in the case of mul ti-taxa samples. With field samples (buffy coat from cattle blood) sensitiv ity was close to the sensitivity observed in single reactions with the clas sical specific primers for the Trypanozoon subgenus and T. congolense-type savannah, but was lower for detection of T. vivax. Additional reaction, imp rovement of DNA preparation, and/or new primers design are necessary to imp rove the sensitivity for detection of T. vivax in field samples. However, t hese primers are suitable for isolate typing through a single PCR. (C) 2001 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd . All rights reserved.