Characterisation of ropy slime-producing Lactobacillus sakei using repetitive element sequence-based PCR

E. Hyytia-Trees, Ulrike Lyhs, H. Korkeala, J. Bjorkroth

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Abstract

Eighteen previously characterised Lactobacillus sakei strains exhibiting varying slime production capabilities in vacuum-packaged meat products were analysed using repetitive element sequence-based PCR (rep-PCR). The single primers BOXA1R and RW3A and the primer pair REP1R-Dt and REP2R-Dt were evaluated for their applicability in L. sakei genotyping. The five different patterns produced by RW3A were the least useful, with the discriminatory power equal to ribotyping. BOXA1R and REP-primer pair both produced six different banding patterns and the combination of these results yielded seven different rep-types. Rep-PCR was concluded to have approximately the same discriminatory power as randomly amplified polymorphic DNA (RAPD) analysis, but was inferior to pulsed-field gel electrophoresis (PFGE). However, if the results of rep-PCR and RAPD were combined, the discrimination was comparable to PFGE, with the exception that within Ribogroup I non-slime-producing strains were indistinguishable from weak slims producers. It was concluded that the combination of the two PCR-based typing techniques, rep-PCR and RAPD, would be a valuable tool in large scale contamination studies at meat processing plants, since results can be obtained rapidly with fewer isolates needing further analysis by PFGE. (C) 1999 Elsevier Science B.V. All rights reserved.
Original languageEnglish
JournalInternational Journal of Food Microbiology
Volume50
Issue number3
Pages (from-to)215-219
ISSN0168-1605
DOIs
Publication statusPublished - 1999
Externally publishedYes

Keywords

  • Food Science
  • Microbiology
  • Contamination studies
  • Rep-PCR
  • Slime-producing L. sakei
  • article
  • bacterium identification
  • food control
  • Lactobacillus sakei
  • meat
  • polymerase chain reaction
  • random amplified polymorphic DNA
  • technique
  • FOOD
  • MICROBIOLOGY
  • RESISTANT STAPHYLOCOCCUS-AUREUS
  • FIELD GEL-ELECTROPHORESIS
  • STREPTOCOCCUS-PNEUMONIAE
  • MEAT-PRODUCTS
  • DNA-SEQUENCES
  • BACTERIA
  • PATTERNS
  • GENOMES
  • slime-producing L. sakei
  • rep-PCR
  • contamination studies
  • food microbiology
  • genotyping
  • meat products vacuum packed
  • Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Regular Nonsporing Gram-Positive Rods [07830] Lactobacillus sakei food pathogen
  • DNA
  • ropy slime
  • 13502, Food technology - General and methods
  • 31500, Genetics of bacteria and viruses
  • 39002, Food microbiology - Food and beverage spoilage and contamination
  • Biochemistry and Molecular Biophysics
  • pulse field gel electrophoresis analytical method
  • RAPD analysis random amplified polymorphic DNA analysis analytical method
  • repetitive element sequence based polymerase chain reaction rep-PCR analytical method
  • Foods
  • Methods and Techniques
  • Molecular Genetics
  • FOOD SAFETY ANIMAL FOODS
  • GENETIC TECHNIQUES
  • LACTOBACILLUS
  • LACTOBACILLUS SAKEI
  • MEAT PRODUCTS
  • NUCLEIC ACIDS
  • PCR
  • RAPD
  • TYPING
  • Hygiene and toxicology
  • Microbiological aspects

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