Construção de iniciadores e otimização de ensaios de pcr e de nested-pcr para a detecção específica de Tritrichomonas foetus
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Data
2008-09
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Resumo
Tritrichomonas foetus is a pathogenic protozoan that causes a venereal disease in cattle known as bovine genital
tricomonosis. In spite of the efficacy to recognize the target genomic DNA, the protocols so far developed for the
diagnosis of this organism by PCR promote some inespecific amplifications or they are unable to discriminate T.
foetus against other species within the genus. The objective of this study was to assess and optimize PCR and
nested-PCR assays for the specific diagnosis of T. foetus, using novel primers selected from the alignment of
sequences of the genes 18S rRNA, 5.8S rRNA, 28S rRNA and of the internal transcribed spacers of the rDNA
(ITS1 and ITS2). A pair of primers was constructed for the genus-specific amplification of a 648 bp fragment and
two others to amplify T. foetus species-specific fragments of 343 and 429 bp. No cross amplification was observed
against Bos taurus genomic DNA neither against the DNA of usual bovine genital pathogens. Both, single and
nested-PCR assays, presented analytical sensitivity to detect at least two T. foetus organisms.
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Reação em cadeia da polimerase, DNAr, Diagnóstico molecular, Tricomonose, Bovinos, Polymerase chain reaction, rDNA, Molecular diagnosis, Trichomonosis, Bovine
Citação
FERNANDES, Paula Rogério; SILVA, Andréa Caetano da; GAMBARINI, Maria Lúcia; LINHARES, Guido Fontgalland Coelho. Construção de iniciadores e otimização de ensaios de pcr e de nested-pcr para a detecção específica de Tritrichomonas foetus. Revista Brasileira de Parasitologia Veterinária, v. 17, n. 3, Jaboticabal, p. 133-138, jul./set. 2008.