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Detection of Leptospira pomona in bovine semen by fluorescent capillary eletrophoresis

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Detalhes bibliográficos
Resumo:This study was performed in order to evaluate the detection limit of PCR with fluorescent capillary electrophoresis for Leptospira pomona diagnosis in bovine semen. Negative bovine semen samples were artificially contaminated with Leptospira pomona (10(0) to 10(7) bacteria/ml) and DNA was extracted by phenol/chloroform protocol. DNA fragments visualization was done by three electrophoresis methods: under UV light in 2% agarose gel, silver staining 8% polyacrylamide gel and fluorescent capillary electrophoresis. The detection limit of capillary electrophoresis for Leptospira pomona was 10²bacteria/ml. Under UV light, in 2% agarose gel, the detection limit was of 10(4) bacteria/ml while for silver stained 8% polyacrylamide gel it was 10² bacteria/ml. PCR with fluorescent capillary electrophoresis is an efficient and rapid diagnostic test for DNA detection of Leptospira in bovine semen and this can be an important tool for herd and semen sanitary control in artificial insemination centers.
Autores principais:Dias, Francisca Elda Ferreira
Outros Autores:Aoki, Sérgio Morais; Mesquita, Lígia Garcia; Nunes, Caris Maroni; Garcia, José Fernando
Assunto:Sêmen Bovino Leptospira PCR Eletroforese Capilar Semen Bovine Leptospira PCR Capillary Eletrophoresis
Ano:2006
País:Brasil
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:IBICT
Idioma:português
Origem:Oasisbr
Descrição
Resumo:This study was performed in order to evaluate the detection limit of PCR with fluorescent capillary electrophoresis for Leptospira pomona diagnosis in bovine semen. Negative bovine semen samples were artificially contaminated with Leptospira pomona (10(0) to 10(7) bacteria/ml) and DNA was extracted by phenol/chloroform protocol. DNA fragments visualization was done by three electrophoresis methods: under UV light in 2% agarose gel, silver staining 8% polyacrylamide gel and fluorescent capillary electrophoresis. The detection limit of capillary electrophoresis for Leptospira pomona was 10²bacteria/ml. Under UV light, in 2% agarose gel, the detection limit was of 10(4) bacteria/ml while for silver stained 8% polyacrylamide gel it was 10² bacteria/ml. PCR with fluorescent capillary electrophoresis is an efficient and rapid diagnostic test for DNA detection of Leptospira in bovine semen and this can be an important tool for herd and semen sanitary control in artificial insemination centers.

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