AI Article Synopsis

  • A novel real-time PCR method was developed for detecting Listeria monocytogenes in food, utilizing hydrolysis probes to provide quicker and simultaneous results.
  • DNA isolation is performed using the BIOTECON foodproof ShortPrep II Kit, followed by analysis with the foodproof Listeria monocytogenes Detection Kit, which includes specific primers, hydrolysis probes, and premixed reagents for accurate results.
  • An internal study tested various food samples (soft cheese, coalfish, smoked ham) with both low and high levels of L. monocytogenes, comparing the PCR results to traditional cultural methods recommended by US food safety authorities.

Article Abstract

A method was developed for the detection of Listeria monocytogenes in food. The method is based on real-time PCR using hydrolysis probes (5' Nuclease). This advanced PCR method was designed to reduce the time necessary to achieve results from PCR reactions and to enable the user to monitor the amplification of the PCR product simultaneously, in real-time. After DNA isolation using the BIOTECON foodproof ShortPrep II Kit designed for the rapid preparation of L. monocytogenes DNA for direct use in PCR, the real-time detection of L. monocytogenes DNA is carried out using the foodproof Listeria monocytogenes Detection Kit. The kit provides primers and hydrolysis probes for sequence-specific detection, convenient premixed reagents, and controls for reliable interpretation of results. For the internal comparison study, three different foods (soft cheese, coalfish, and smoked ham) were analyzed, chosen from the 15 food groups recommended by the AOAC Research Institute for detection of L. monocytogenes. From each food, 20 samples were inoculated with a low level (1-10 CFU/25 g) and 20 samples with a high level (10-50 CFU/25 g) of L. monocytogenes. Additionally, five nonspiked samples were prepared from each food. Depending on the matrix, the food samples were examined with the test kits and compared with the cultural methods according to the U.S. Food and Drug Administration's Bacteriological Analytical Manual or the U.S. Department of Agriculture/Food Safety and Inspection Service Microbiology Laboratory Guidebook.

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http://dx.doi.org/10.5740/jaoacint.11-0096DOI Listing

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