Sensitivity of different Trypanosoma vivax specific primers for the diagnosis of livestock trypanosomosis using different DNA extraction methods.
There are several T. vivax specific primers developed for PCR diagnosis. Most of these primers were validated under different DNA extraction methods and study designs leading to heterogeneity of results. The objective of the present study was to validate PCR as a diagnostic test for T. vivax trypanosomosis by means of determining the test sensitivity of different published specific primers with different sample preparations. Four different DNA extraction methods were used to test the sensitivity of PCR with four different primer sets. DNA was extracted directly from whole blood samples, blood dried on filter papers or blood dried on FTA cards. The results showed that the sensitivity of PCR with each primer set was highly dependant of the sample preparation and DNA extraction method. The highest sensitivities for all the primers tested were determined using DNA extracted from whole blood samples, while the lowest sensitivities were obtained when DNA was extracted from filter paper preparations. To conclude, the obtained results are discussed and a protocol for diagnosis and surveillance for T. vivax trypanosomosis is recommended.
Gonzales
J L
JL
Laboratorio de Investigación y Diagnóstico Veterinario "LIDIVET", Av. Ejercito Nacional 153, P.O. Box 29, Santa Cruz, Bolivia. pepevet@yahoo.com
Loza
A
A
Chacon
E
E
D000818:Animals; D001838:Bolivia; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry / Q000592:standards; D016054:DNA, Protozoan / Q000097:blood / Q000302:isolation & purification*; D008297:Male; D018512:Parasitemia / Q000469:parasitology / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood / Q000175:diagnosis* / Q000469:parasitology; D016689:Trypanosoma vivax / Q000235:genetics / Q000302:isolation & purification*; D014352:Trypanosomiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D014354:Trypanosomiasis, Bovine / Q000469:parasitology
D017931:DNA Primers; D016054:DNA, Protozoan