PCR detection of Bacillus and Staphylococcus in various foods

Journal of Food Protection
Shigeru NakanoToshihiro Yamada

Abstract

A broad-range PCR assay for the detection of bacteria belonging to Bacillus and Staphylococcus genera was developed. Primers targeting the bacterial 16S rRNA gene were newly designed and used in a PCR assay. To determine the specificity of the assay, 81 different bacterial strains (of 50 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for every tested Bacillus, Staphylococcus, or Aerococcus strain. In addition, the result for Listeria grayi was positive with lower PCR product. For all other bacterial strains and eukaryotes tested, results were negative. Bacterial DNA was prepared with the use of achromopeptidase and Chelex 100 resin from culture after growth in brain heart infusion medium. To test the sensitivity of this PCR assay for Bacillus or Staphylococcus genus, either Bacillus cereus or Staphylococcus aureus was inoculated into various foods with undetectable levels of endogenous microbial contamination as an indicator. Inoculation of bacteria at 10 to 30 CFU/g of food was followed by a 5-h enrichment culture step after which the PCR assay allowed the detection of bacterial cells. When the inoculation (B. cereus or S. aureus) of 10 to 90 CFU/g into noodle foods containing endogenous microflor...Continue Reading

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Oct 24, 2003·Journal of Food Protection·Shigeru NakanoToshihiro Yamada

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Citations

Jan 13, 2005·Canadian Journal of Microbiology·Kiev S Gracias, John L McKillip
Jan 30, 2007·Applied and Environmental Microbiology·Martina FrickerMonika Ehling-Schulz
May 5, 2017·ACS Central Science·Qifan ZhangTimothy M Swager

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