Skip to main content
Log in

DNA sequence of a Salmonella-specific DNA fragment and the use of oligonucleotide probes for Salmonella detection

  • Applied Genetics and Regulation
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Summary

Hybridization specificity of a 1.8-kb HindIII DNA fragment isolated from Salmonella typhimurium by a molecular cloning technique was confirmed by colony hybridization with 327 Salmonella isolates of various serotypes and 56 non-Salmonella isolates including Enterobacteriaceae closely related to Salmonella, such as Escherichia coli, Klebsiella, Citrobacter and Shigella. It was found that this 1.8-kb DNA fragment was highly specific for all the Salmonella isolates tested. The DNA sequence of this 1.8-kb fragment was then determined by the dideoxynucleotide chain termination method. According to this DNA sequence, six oligonucleotide fragments ranging from 17- to 26-mer were then chemically synthesized and tested for their hybridization specificities. Results show that three of the six oligonucleotide fragments are highly specific for all 327 Salmonella strains tested and can be used as probes for the specific detection of Salmonella in foods or other samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Denhart DT (1966) A membrane-filter technique for detection of complementary DNA. Biochem Biophys Res Commun 23:641–646

    Article  Google Scholar 

  • Fitts R (1985) Development of DNA-DNA hybridization test for the presence of Salmonella in food. Food Technol 39:95–102

    CAS  Google Scholar 

  • Fitts R, Diamond C, hamilton C, Neri M (1983) DNA-DNA hybridization assay for detection of Salmonella spp. in foods. Appl Environ Microbiol 46:1146–1151

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hill WE (1981) DNA hybridization method for detection of enterotoxigenic Escherichia coli in human isolates and possible application to food samples. J Food Safety 3:233–247

    Article  Google Scholar 

  • Hill WE, Payne WL, Aulisio CCG (1983) Detection and enumeration of virulent Yersinia enterocolitica in food by DNA colony hybridization. Appl Environ Microbiol 46:636–641

    CAS  PubMed  PubMed Central  Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Spring Spring Harbor Laboratory, Cold Spring Harbor, N. Y., pp 124–125

    Google Scholar 

  • Moseley SL, Hug AR, Alim MA, Samadpour-Motolebi M, Falkow S (1980) Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis 142:892–898

    Article  CAS  Google Scholar 

  • Notermans S, Meuvelman KJ, Wernars K (1988) Synthetic enterotoxin B DNA probe for detection of enterotoxigenic Staphylococcus aureus strains. Appl Environ Microbiol 54:531–533

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sanger F, Nicklen S, Calson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Article  CAS  Google Scholar 

  • Sanger F, Coulson AR, Barrell BG, Smith AJH, Roe BA (1980) Cloning on single stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178

    Article  CAS  Google Scholar 

  • Swaminathan B, Aleixo JAG, Minnich SA (1985) Enzyme immunoassays for Salmonella: one day testing is now reality. Food Technol 39:83–89

    CAS  Google Scholar 

  • Tsen HY, Chen MH, Shieh JS, Wang SJ, Hu NT (1989) Possible use of a 1.8 kb DNA fragment for the specific detection of Salmonella in foods. J Ferment Bioeng 68:1–6

    Article  CAS  Google Scholar 

  • US Food and Drug Administration (1978) Bacteriological analytical manual, 5th edn. Association of Analytical Chemists, Arlington, VA

    Google Scholar 

  • Wirth D, Pratt DM (1982) Rapid identification of Leishmania species by specific hybridization of kintoplast DNA in cutaneous lesions. Proc Natl Acad Sci USA 79:6999–7003

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Offprint requests to: H.-Y. Tsen

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tsen, HY., Wang, SJ., Roe, B.A. et al. DNA sequence of a Salmonella-specific DNA fragment and the use of oligonucleotide probes for Salmonella detection. Appl Microbiol Biotechnol 35, 339–347 (1991). https://doi.org/10.1007/BF00172723

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00172723

Keywords

Navigation