Trypanosoma evansi: A comparison of PCR and parasitological diagnostic tests in experimentally infected mice.
Trypanosoma evansi is the causative agent of equine trypanosomosis, disease that affects horse's productivity and health. Parasitological and molecular methods are mostly used to detect the infection. The aim of this work was evaluate PCR sensitivity to detect T. evansi using the primers 21/22-mer, ITS1, ESAG 6/7 and TBR 1/2 designed from repetitive (multicopies) genomic sequences. The results were compare with two parasitological tests in mice, micro-haematocrite centrifugation technique and direct microscopic examination. The results shows (a) that the minimum amount of DNA from blood of highly parasitaemic mice that was detectable by PCR was 0.001 ng, using the ESAG6/7 and TBR1/2 primer, (b) using TBR1/2 primer for parasites purified could detect 0.000001 ng and (c) in the prepatent period PCR detect the presence of parasites earlier than parasitological techniques. Nevertheless, the percentage of detection for PCR varies depending on primer employed with 60% and 66% for ITS1 and 21/22-mer, and 80% for ESAG6/7 and TBR1/2. Consequently, TBR1/2 and ESAG6/7 were the best primers to monitor T. evansi infections in mice. For epidemiological application, such comparative evaluation should be made for detection of T. evansi in livestock such as horses.
Universidad Nacional Experimental Simón Rodríguez, Instituto de Estudios Científicos y Tecnológicos, Centro de Estudios Biomédicos y Veterinarios, Apartado Postal 47925, Caracas 1041A, Venezuela.
D
D
Fernández
B
B
González-Baradat
M
M
Eleizalde
E
E
González-Marcano
T
T
Perrone
M
M
Mendoza
D000818:Animals; D002417:Cattle; D002498:Centrifugation; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis / Q000097:blood; D004195:Disease Models, Animal; D018397:Equidae; D005260:Female; D006734:Horse Diseases / Q000175:diagnosis / Q000469:parasitology; D006736:Horses; D051379:Mice; D018512:Parasitemia / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000592:standards / Q000662:veterinary*; D012377:Rodentia; D012680:Sensitivity and Specificity; D014345:Trypanosoma / Q000235:genetics / Q000302:isolation & purification*; D014352:Trypanosomiasis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*
D017931:DNA Primers; D016054:DNA, Protozoan