Development of a new multiplex PCR for the detection of Clostridium spp., responsible for the late blowing in cheese (Abstract/Poster in atti di convegno)

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  • Development of a new multiplex PCR for the detection of Clostridium spp., responsible for the late blowing in cheese (Abstract/Poster in atti di convegno) (literal)
Anno
  • 2009-01-01T00:00:00+01:00 (literal)
Alternative label
  • Laura Vanoni; Paola Cremonesi; Milena Brasca; Roberta Lodi; Tiziana Silvetti (2009)
    Development of a new multiplex PCR for the detection of Clostridium spp., responsible for the late blowing in cheese
    in Second SAFE Consortium International Congress on Food Safety NOVEL TECHNOLOGIES AND FOOD QUALITY, SAFETY AND HEALTH, Girona, Catalunya, Spain, 27-29 aprile 2009
    (literal)
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  • Laura Vanoni; Paola Cremonesi; Milena Brasca; Roberta Lodi; Tiziana Silvetti (literal)
Pagina inizio
  • 150 (literal)
Pagina fine
  • 151 (literal)
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  • ISSN: 1819-7779 (literal)
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  • Second SAFE consortium International Congress on Food Safety (literal)
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  • DAL 27 AL 29 APRILE 2009 (literal)
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  • 2 (literal)
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  • Late blowing, caused by the outgrowth of clostridial spores present in raw milk and originating from silage, can create considerable product loss, especially in the production of hard and semi-hard cheeses. The conventional method for the isolation of Clostridium spp. from cheeses with late-blowing symptoms is very complicated and the identification of isolates is problematic. The aim of this work was the development of a multiplex PCR method for the detection of the main dairy-related clostridia such as: Cl. beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricum. Genomic DNA from five reference strains, Cl. baratii DSM 601, Cl. beijerinckii DSM 791, Cl. butyricum DSM 10702, Cl. sporogenes ATCC 3584, Cl. tyrobutyricum DSM 2637, was extracted following the protocol described in the literature. All PCR primer were designed using the Primer3 programme (http:// www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi). All the primer were chosen on the basis of similar melting temperatures and minimal interactions, and resulted in differently sized products distinguishable in agarose gel eletrophoresis. The specificity of the primer was tested by individually analysing each primer pair and the primer pair combined in the multiplex PCR. Samples derived from silage, raw milk and hard cheese were analysed by the most probable number (MPN) enumeration. Forty-eight bacterial strains isolated from gas positive tubes were used to check the reliability of the multiplex PCR assay. It is interesting to note that the samples not identified by the multiplex PCR assay were amplified by V2-V3 16S rRNA primer pair and the sequencing revealed the aligned 16S rRNA sequences to be Paenibacillus spp. This method provides a simple promising alternative to traditional microbiological methods for a rapid, sensitive detection of clostridia in dairy products. (literal)
Note
  • Poster (literal)
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  • Institute of Sciences of Food Production - Italian National Research Council, Milan, Italy Institute of Agricultural Biology and Biotechnology -Italian National Research Council, Milan, Italy (literal)
Titolo
  • Development of a new multiplex PCR for the detection of Clostridium spp., responsible for the late blowing in cheese (literal)
Abstract
  • Late blowing, caused by the outgrowth of clostridial spores present in raw milk and originating from silage, can create considerable product loss, especially in the production of hard and semi-hard cheeses. The conventional method for the isolation of Clostridium spp. from cheeses with late-blowing symptoms is very complicated and the identification of isolates is problematic. The aim of this work was the development of a multiplex PCR method for the detection of the main dairy-related clostridia such as: Cl. beijerinckii, Cl. butyricum, Cl. sporogenes, Cl. tyrobutyricum. Genomic DNA from five reference strains, Cl. baratii DSM 601, Cl. beijerinckii DSM 791, Cl. butyricum DSM 10702, Cl. sporogenes ATCC 3584, Cl. tyrobutyricum DSM 2637, was extracted following the protocol described in the literature. All PCR primer were designed using the Primer3 programme (http:// www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi). All the primer were chosen on the basis of similar melting temperatures and minimal interactions, and resulted in differently sized products distinguishable in agarose gel eletrophoresis. The specificity of the primer was tested by individually analysing each primer pair and the primer pair combined in the multiplex PCR. Samples derived from silage, raw milk and hard cheese were analysed by the most probable number (MPN) enumeration. Forty-eight bacterial strains isolated from gas positive tubes were used to check the reliability of the multiplex PCR assay. It is interesting to note that the samples not identified by the multiplex PCR assay were amplified by V2-V3 16S rRNA primer pair and the sequencing revealed the aligned 16S rRNA sequences to be Paenibacillus spp. This method provides a simple promising alternative to traditional microbiological methods for a rapid, sensitive detection of clostridia in dairy products. (literal)
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