Development of a TaqMan PCR assay for the detection of Trypanosoma evansi, the agent of surra.
A TaqMan PCR assay was developed for the detection of Trypanosoma evansi. The assay targets the internal transcribed spacer 1 (ITS-1) region of rRNA. The ITS-1 region of eleven strains of T. evansi from widely separated geographical regions were sequenced and alignments compared. Primers and probe for the test were designed from these sequence data. The assay was tested using blood from infected rats and was found to be sensitive, detecting less than one genomic equivalent of T. evansi. The assay has been tested against 10 different species of trypanosomes found in native animals in Australia and did not detect any of these trypanosome species. Time course experiments using rats infected with T. evansi were performed to compare the TaqMan assay with the Haematocrit centrifugation test (HCT) and the mouse inoculation (MI) assay. The assay was more sensitive than the HCT but not as sensitive as the MI. The TaqMan assay has the ability to rapidly detect T. evansi and determine the number of organisms present in a blood sample from an infected animal. This is the first time a TaqMan assay has been developed for the detection of T. evansi.
Taylor
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CSIRO Livestock Industries, Australian Animal Health Laboratory, Private Bag 24, Geelong, Victoria 3220, Australia. Trevor.Taylor@csiro.au
Boyle
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Bingham
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D000820:Animal Diseases / Q000175:diagnosis* / Q000469:parasitology; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D016000:Cluster Analysis; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004285:Dogs; D006801:Humans; D008394:Marsupialia; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity; D019914:Taq Polymerase; D014345:Trypanosoma / Q000302:isolation & purification*; D014352:Trypanosomiasis / Q000175:diagnosis* / Q000469:parasitology
D021903:DNA, Ribosomal Spacer; D016053:RNA, Protozoan; D012335:RNA, Ribosomal; D019914:Taq Polymerase