title: PCR detection of psychrotolerant clostridia associated with deep tissue spoilage of vacuum-packed chilled meats.
abstract: OBJECTIVE
  To develop a practical molecular procedure that directly (without isolation) and specifically detects the presence of clostridia, which cause the deep tissue spoilage condition.
  
  
  RESULTS
  A primer set was designed and a PCR amplification procedure developed to detect the presence of Clostridium algidicarnis and Cl. putrefaciens 16S rDNA gene fragments in meat. The procedure yielded amplicons of the expected size with homologous DNA templates, but failed to give PCR products with DNAs from 47 food clostridia and common meat spoilage micro-organisms. The minimum level of detection was 10(4) cfu g-1 for nonenriched meat samples. Based on the established specificity of these primers, as well as DNA sequencing of amplicons, the presence of Cl. algidicarnis and/or Cl. putrefaciens was confirmed in a swab sample taken from the cartilage of an ovine stifle joint, which on opening exhibited strong offensive odours.
  
  
  CONCLUSIONS
  The developed method can be used for rapid detection of clostridia causing deep tissue spoilage in commercial vacuum packs.
  
  
  CONCLUSIONS
  The paper reports practical procedures that can be used for rapid confirmation of the causative agents of deep tissue clostridial spoilage in commercial vacuum-packed chilled meats.
authors.affiliation: AgResearch, Hamilton, New Zealand.
authors.forename: J A
authors.initials: JA
authors.lastname: Boerema
authors.forename: D M
authors.initials: DM
authors.lastname: Broda
authors.forename: R G
authors.initials: RG
authors.lastname: Bell
mesh_terms: D000818:Animals; D003013:Clostridium / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D003080:Cold Temperature; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D005516:Food Microbiology; D018857:Food Packaging; D008461:Meat Products / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D012756:Sheep; D014618:Vacuum
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal