The detection of non-RoTat 1.2 Trypanosoma evansi

• Published in: Experimental parasitology Vol. 110; no. 1; pp. 30 - 38
• Main Authors: Ngaira, J.M., Olembo, N.K., Njagi, E.N.M., Ngeranwa, J.J.N.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.05.2005; Elsevier
• Subjects: African trypanosomiasis; Amino Acid Sequence; amino acid sequences; Animals; Base Sequence; Biological and medical sciences; Camelus - parasitology; Cloning, Molecular; complementary DNA; detection; Diagnosis; disease diagnosis; DNA, Complementary - chemistry; DNA, Complementary - isolation & purification; DNA, Protozoan - chemistry; DNA, Protozoan - isolation & purification; Fundamental and applied biological sciences. Psychology; Kenya; Life cycle. Host-agent relationship. Pathogenesis; Molecular Sequence Data; nucleotide sequences; polymerase chain reaction; Protozoa; Restriction Mapping - veterinary; Reverse Transcriptase Polymerase Chain Reaction - veterinary; RNA, Protozoan - chemistry; RNA, Protozoan - isolation & purification; Sensitivity and Specificity; Sequence Alignment - veterinary; Trypanosoma - genetics; Trypanosoma - immunology; Trypanosoma - isolation & purification; Trypanosoma brucei; Trypanosoma congolense; Trypanosoma evansi; Trypanosoma vivax; Trypanosomiasis, African - diagnosis; Trypanosomiasis, African - parasitology; Trypanosomiasis, African - veterinary; Variant surface glycoprotein; variant surface glycoproteins; Variant Surface Glycoproteins, Trypanosoma - genetics; Kenya; Africa; Trypanosoma evansi; Diagnosis; Variant surface glycoprotein; Kenya; Kinetoplastida; Variant; Protozoa; Trypaiosama evansi; Glycoprotein; Parasite; Detection
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2005.01.001

The majority of Trypanosoma evansi can be detected using diagnostic tests based on the variant surface glycoprotein (VSG) of Trypanosoma evansi Rode Trypanozoon antigen type (RoTat) 1.2. Exceptions are a number of T. evansi isolated in Kenya. To characterize T. evansi that are undetected by RoTat 1.2, we cloned and sequenced the VSG cDNA from T. evansi JN 2118Hu, an isolate devoid of the RoTat 1.2 VSG gene. A 273 bp DNA segment of the VSG gene was targeted in PCR amplification for the detection of non-RoTat 1.2 T. evansi. Genomic DNA samples from different trypanosomes were tested including 32 T. evansi, 10 Trypanosoma brucei, three Trypanosoma congolense, and one Trypanosoma vivax. Comparison was by PCR amplification of a 488 bp fragment of RoTat1.2 VSG gene. Results showed that the expected 273 bp amplification product was present in all five non-RoTat 1.2 T. evansi tested and was absent in all 27 RoTat 1.2-positive T. evansi tested. It was also absent in all other trypanosomes tested. The PCR test developed in this study is specific for non-RoTat 1.2 T. evansi.