Sample preparation methods for PCR detection of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on beef chuck shoulder using a single enrichment medium

  • Bhaduri S
  • Cottrell B
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Abstract

To improve the utility of the polymerase chain reaction (PCR) for food samples, methods for prepg. template DNA were developed to remove PCR inhibitors. Beef chuck shoulder medallions, artificially contaminated, individually or in combination, with Escherichia coli serotype O157:H7 strain FSIS 45753-35, Salmonella typhimurium DT104 strain 13HP, or Listeria monocytogenes strain Scott A at concns. of 10, 1 and 0.5 cfu/cm2 were swabbed with a sponge, and the sponges were enriched for 18 h at 37 DegC in universal pre-enrichment broth (UPB). Enriched broth cultures (EBC), cell pellets (CP), or phosphate-buffered saline-washed cell pellets (PBSCP) from enriched sponge samples were compared for detection of E. coli O157:H7, S. typhimurium DT104, or L. monocytogenes by the PCR using the BAX system. Recovery of the three organisms was effective for detection of each pathogen at initial levels of 10, 1 and 0.5 cfu/cm2 when inoculated sep., or in combination, onto the beef samples. Use of EBC, CP, or PBSCP of sponge-swabbed samples eliminated problems assocd. with inhibition of the PCR by food components, time-consuming extn. of DNA, and inhibition due to large amts. of non-target DNA derived from the food. The procedure involving enrichment of sponge-swabbed beef samples in UPB followed by PCR amplification using EBC with the BAXTMsystem is the most efficient and simple method for detection of E. coli O157:H7, S. typhimurium DT104, and L. monocytogenes. (c) 2001 Academic Press.

Author-supplied keywords

  • Culture media (universal pre-enrichment broth
  • DNA Role: ANT (Analyte), FFD (Food or feed use), A
  • Escherichia coli
  • Escherichia detection beef enrichment medium spong
  • Food analysis
  • Listeria beef enrichment medium sponge swab PCR
  • Listeria monocytogenes (sample prepn. methods for
  • Meat (beef, artificially contaminated
  • PCR (BAX system
  • Salmonella detection beef enrichment medium sponge
  • Salmonella typhimurium (DT104
  • Sponges (procedure involving enrichment of sponge-
  • food analysis bacterium sponge swab UPB broth PCR
  • procedure involving enrichment of sponge-swabbed b
  • sample prepn. methods for PCR detection of Escheri

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Authors

  • S Bhaduri

  • B Cottrell

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