Improved method for rapid DNA extraction of Mastitis pathogens directly from milk (Articolo in rivista)

Type
Label
  • Improved method for rapid DNA extraction of Mastitis pathogens directly from milk (Articolo in rivista) (literal)
Anno
  • 2006-01-01T00:00:00+01:00 (literal)
Alternative label
  • Paola Cremonesi; Bianca Castiglioni; Giulia Malferrari; Ida Biunno; Chiara Vimercati; Paolo Moroni; Stefano Morandi; Massimo Luzzana (2006)
    Improved method for rapid DNA extraction of Mastitis pathogens directly from milk
    in Journal of dairy science
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Paola Cremonesi; Bianca Castiglioni; Giulia Malferrari; Ida Biunno; Chiara Vimercati; Paolo Moroni; Stefano Morandi; Massimo Luzzana (literal)
Pagina inizio
  • 163 (literal)
Pagina fine
  • 169 (literal)
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  • 89 (literal)
Rivista
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  • 7 (literal)
Note
  • PubMe (literal)
  • ISI Web of Science (WOS) (literal)
  • Scopus (literal)
  • Google Scholar (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • Department of Biomedical Sciences and Technologies, University of Milan, Segrate, Milan, Italy Institute of Agricultural Biology and Biotechnology-Italian National Research Council, Milan, Italy Center for Bio-Molecular Interdisciplinary Studies and Industrial Applications (CISI), University of Milan, Segrate, Milan, Italy Institute of Biomedical Technologies-Italian National Research Council, Segrate, Milan, Italy # Department of Animal Pathology, Hygiene and Veterinary Public Health University of Milan, Italy || Institute of Sciences of Food Production-Italian National Research Council, Milan, Italy (literal)
Titolo
  • Improved method for rapid DNA extraction of Mastitis pathogens directly from milk (literal)
Abstract
  • Efficient control against bovine mastitis requires sensitive, rapid and specific tests to detect and identify the main bacteria that cause heavy losses to the dairy industry. Molecular detection of pathogenic microorganisms is based on DNA amplification of the target pathogen; therefore, an efficient extraction of DNA from pathogenic bacteria is a major step. In this study, we aimed to develop a specific, sensitive and fast method to extract DNA from the main Gram positive bacteria known to cause bovine mastitis (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis) directly from milk samples. The DNA extraction method is based on the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate together with the nucleic acid-binding properties of the silica particles. An efficient protocol consisting of six total basic steps (three of which were done twice) was developed and applied directly to milk samples. The absence of PCR inhibitors and the DNA quality were evaluated by PCR amplification of the species-specific DNA sequences of the target bacteria. The level of sensitivity achieved in our experiments is applicable to milk sample analysis without sample enrichment. (literal)
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