Uso de Chelex-100 para o diagnóstico molecular de cinco patógenos animais
DOI:
https://doi.org/10.14409/favecv.v20i1.9724Palavras-chave:
Veterinary, Diagnosis, DNA, Chelex-100, PCRResumo
As técnicas moleculares têm contribuído para melhorar o diagnóstico veterinário tradicional e a extração de ácido nucléico é crucial. O objetivo deste trabalho foi comparar o método de extração de DNA Chelex-100 (M1) com papel Whatman (M2), fenol-clorofórmio (M3) ou kits comerciais (M4), e determinar um método sensível e de baixo custo para o diagnóstico de patógenos animais economicamente ou zoonoticamente relevantes recebendo pouca atenção. Da urina, órgãos, sêmen, sangue e mucosa intestinal, foi extraído DNA das bactérias Leptospira interrogans sorovar Pomona Pomona (com M1 e M2), Brucella melitensis (com M1, M3 e M4), Salmonella ser. Abortusequi (M1 e M4), e da Leishmania spp. (M1, M3 e M4) e Eimeria spp. (M1 e M3), respectivamente. A sensibilidade dos protocolos foi analisada por PCR. O método M1 demonstrou sensibilidade semelhante para S. Abortusequi, Leishmania spp. e Eimeria spp., sendo melhor para L. interrogans e ligeiramente inferior para B. melitensis. Pela primeira vez, um método simples e barato de extração de DNA foi utilizado com sucesso nesses patógenos veterinários, especialmente importante em laboratórios com poucos recursos econômicos, contribuindo para o diagnóstico de leptospirose, brucelose, leishmaniose e coccidiose, bem como para a epidemiologia molecular de salmonelose em amostras de sêmen equino.Referências
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