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    Rapid identification of isometamidium-resistant stocks of Trypanosoma b. brucei by PCR-RFLP (2006)

    Art
    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Autoren
    Afework, Y.
    Mäser, P.
    Etschmann, B.
    von Samson-Himmelstjerna, G
    Zessin, K. H.
    Clausen, P. H.
    Quelle
    Parasitology research; 99(3) — S. 253–261
    ISSN: 0932-0113
    Sprache
    Englisch
    Verweise
    Pubmed: 16541260
    Kontakt
    Institut für Veterinär-Physiologie

    Oertzenweg 19 b
    14163 Berlin
    Tel.+49 30 838 62600 Fax.+49 30 838-62610
    email:physiologie@vetmed.fu-berlin.de

    Abstract / Zusammenfassung

    Analyses were made on the adenosine transporter-1 gene in Trypanosoma brucei (TbAT1), encoding a P2-like nucleoside transporter, from T. brucei brucei field stocks to investigate a possible link between the presence of mutations in this gene and isometamidium resistance. We have analysed the gene from 11 isometamidium-sensitive field stocks isolated from cattle in Uganda, two sensitive reference clones and two resistant reference clones. Sequence alignment showed that the isometamidium-sensitive T. b. brucei contained the wild-type sequence patterns. In contrast, the isometamidium-resistant T. b. brucei stocks showed the mutant-type sequence patterns with six point mutations that had previously been reported in a laboratory-derived arsenical-resistant T. brucei strain. In order to analyse the RFLP pattern of a fragment of TbAT1 (nucleotides 430-1108), the 677 bp PCR products from eight of the isometamidium-sensitive and two of the isometamidium-resistant T. b. brucei were subjected to digestion with Sfa NI. The results revealed two different banding patterns: the digest resulted in fragment sizes of 566 and 111 bp in the case of TbAT1 from isometamidium-sensitive stocks, whereas it produced fragment sizes of 435 and 242 bp in the case of TbAT1 from isometamidium-resistant stocks. Thus, the isometamidium-sensitive and resistant T. b. brucei could be successfully distinguished by digestion with the restriction endonuclease Sfa NI.