Application of random amplified polymorphic DNA analysis for detection of Salmonella spp. in foods

Takahisa Miyamoto, Huai Ze Tian, Takashi Okabe, Sudsai Trevanich, Kaoru Asoh, Satoshi Tomoda, Ken-ichi Honjoh, Shoji Hatano

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

The random amplified polymorphic DNA (RAPD) band patterns from 23 Salmonella spp. produced by use of an oligonucleotide primer (called du primer) designed on the basis of the N-terminal sequence of dulcitol 1-phosphate dehydrogenase (5'-GTGGTGACCCAGGATGGCCAGGTG-3') were different from those from 16 non-Salmonella spp. The bands at 460 and 700 bp were produced in all Salmonella strains tested. These RAPD fragments obtained from Salmonella typhimurium strongly hybridized with the corresponding RAPD bands from the other strains of Salmonella, but not with those from non-Salmonella spp. in Southern blot analysis. The RAPD bands were detected by ethidium bromide staining even when genomic DNA prepared from as few as 2.8 X 103 cells was used. The minimum detectable cell number in the initial inoculum of S. typhimurium was 4 X 10-1 CFU/25 g of raw beef after the preenrichment in Enterobacteriaceae enrichment mannitol (EEM) broth for 6 h and the selective enrichment in dulcitol-magnesium chloride-pyridinesulfonic acid-brilliant green-novobiocin (DMPBN) medium for 18 h at 42°C. Seven raw foods inoculated with S. typhimurium at numbers from 4 X 10-1 to 2.6 X 102 CFU/25 g of food were positive in both the RAPD analysis and the conventional culture method.

Original languageEnglish
Pages (from-to)785-791
Number of pages7
JournalJournal of Food Protection
Volume61
Issue number7
DOIs
Publication statusPublished - Jan 1 1998

All Science Journal Classification (ASJC) codes

  • Food Science
  • Microbiology

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