Detection and identification of Trypanosoma of African livestock through a single PCR based on internal transcribed spacer 1 of rDNA.
Primers hybridising with the rDNA cistron have previously been evaluated for PCR diagnosis specific for kinetoplastids, and shown to detect and differentiate the Trypanosoma brucei complex and Trypanosoma cruzi. Kin1 and Kin2 primers, amplifying internal transcribed spacer 1, were subsequently evaluated for the diagnosis of African livestock trypanosomosis. Based on the size of the PCR products obtained, Kin primers allowed detection and identification of three Trypanosoma congolense types (savannah, forest and Kenya Coast), 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 shown to be suitable for the sensitive and type-specific diagnosis of African livestock trypanosome isolates through a single PCR even in the case of multi-taxa samples. With field samples (buffy-coat from cattle blood) sensitivity was close to the sensitivity observed in single reactions with the classical specific primers for the Trypanozoon subgenus and T. congolense-type savannah, but was lower for detection of T. vivax. Additional reaction, improvement of DNA preparation, and/or new primers design are necessary to improve the sensitivity for detection of T. vivax in field samples. However, these primers are suitable for isolate typing through a single PCR.
Desquesnes
M
M
CIRAD-EMVT BP 5035, 34032, Montpellier, France. marc.desquesnes@cirad.fr
McLaughlin
G
G
Zoungrana
A
A
Dávila
A M
AM
D000818:Animals; D002050:Burkina Faso; D002417:Cattle; D017931:DNA Primers; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D014345:Trypanosoma / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D014354:Trypanosomiasis, Bovine / Q000175:diagnosis*
D017931:DNA Primers; D016054:DNA, Protozoan; D021903:DNA, Ribosomal Spacer