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Genet Mol Res ; 14(2): 5103-11, 2015 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-26125702

RESUMO

Single nucleotide polymorphisms at codons 167, 198, and 200 in the ß-tubulin isotype 1 gene have been associated with benz-imidazole resistance. Until now, the only mutation observed in Ancy-lostoma caninum was at codon 200 of this gene. However, the standard-ized methodologies used to detect mutations in this species are faulty. The objective of this study was to standardize a molecular technique based on amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR) for detecting the mutation at codon 200 in the A. caninum ß-tubulin isotype 1 gene. Controls were synthesized both for the absence of the mutation, using conventional PCR, and for the presence of the mutation, using the Megaprimer-PCR technique. After standardization of the ARMS-PCR using the controls, the technique was validated through an analysis of 75 A. caninum DNA samples, fol-lowed by sequencing. The results revealed that the developed technique has high sensitivity, specificity, and reproducibility, which allow its ap-plication in the field.


Assuntos
Ancylostoma/genética , Resistência a Medicamentos/genética , Mutação , Reação em Cadeia da Polimerase/normas , Polimorfismo de Nucleotídeo Único , Tubulina (Proteína)/genética , Ancylostoma/efeitos dos fármacos , Animais , Anti-Helmínticos/farmacologia , Benzimidazóis/farmacologia , Códon , Reação em Cadeia da Polimerase/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA
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