Detection of African animal trypanosomes: the haematocrit centrifugation technique compared to PCR with samples stored on filter paper or in DNA protecting buffer.
The present study aimed at comparing the trypanosome specific 18S-PCR-RFLP using samples stored either on Whatman filter papers (PCR-RFLP-fp) or in a commercial cell lysis and DNA protecting buffer (PCR-RFLP-pb) with the haematocrit centrifugation technique (HCT), a method widely used for the diagnosis of African Animal Trypanosomosis. Out of 411 head of cattle, 49 (11.92%) (CI=8.95-15.45) scored positive for the presence of trypanosomes by HCT whereas 75 (18.25%) (CI=14.63-22.33) and 124 (30.17%) (CI=25.77-34.86) scored positive using PCR-RFLP-fp and PCR-RFLP-pb, respectively. Out of the 49 positives by HCT, 14 (28.57%) (CI=16.58-43.26) and 28 (57.14%) (CI=42.21-71.18) were concordant by PCR-RFLP-fp and PCR-RFLP-pb, respectively. None of the PCR techniques detected parasites from the Trypanozoon group. Although HCT detected more cases of Trypanosoma vivax (33), species identification using PCR-RFLP-fp and PCR-RFLP-pb were significantly different (p<0.001) from the HCT technique. The use of DNA protective buffer is thus recommended as the output of the PCR-RFLP-pb is improved and the risk of contamination between samples is reduced.
Department of Microbiology and Veterinary Public Health, Jimma University, College of Agriculture and Veterinary Medicine, P.O. Box 307, Jimma, Ethiopia.
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Y
Moti
College of Veterinary Medicine and Agriculture, Addis Ababa University, P.O. Box 34, Debre Zeit, Ethiopia.
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R
Fikru
Institute of Tropical Medicine, Department of Biomedical Sciences - Systems Biology, Nationalestraat 155, Antwerp, Belgium.
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Büscher
Institute of Tropical Medicine, Department of Biomedical Sciences - Systems Biology, Nationalestraat 155, Antwerp, Belgium.
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J
Van Den Abbeele
Universiteit Gent, Faculty of Veterinary Sciences - Comparative Physiology and Biometrics, Salisburylaan 133, B-9820 Merelbeke, Belgium.
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Duchateau
Institute of Tropical Medicine, Department of Biomedical Sciences - Systems Biology, Nationalestraat 155, Antwerp, Belgium. Electronic address: vdelespaux@itg.be.
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Delespaux
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D002498:Centrifugation / Q000592:standards / Q000662:veterinary*; D005002:Ethiopia; D005260:Female; D006400:Hematocrit / Q000592:standards / Q000662:veterinary*; D008297:Male; D016133:Polymerase Chain Reaction / Q000592:standards / Q000662:veterinary*; D014345:Trypanosoma / Q000235:genetics / Q000502:physiology*; D014353:Trypanosomiasis, African / Q000175:diagnosis / Q000662:veterinary*; D014730:Veterinary Medicine / Q000379:methods*
Buffer; Cattle; Ethiopia; Trypanosoma; Trypanosoma congolense; Trypanosoma vivax