Development of a novel approach for the production of dried genomic DNA for use as standards for qualitative PCR testing of food-borne pathogens

S. Trapmann, P. Catalani, Jeffrey Hoorfar, J. Prokisch, P. Iwaarden, H. Schimmel

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

As part of a multi-centre European project, FOOD-PCR, the feasibility of a novel approach for production of dried bacterial DNA that could be used as certified reference materials (CRM) was assessed. Selected strains of Salmonella typhimurium, Listeria monocytogenes, Escherichia coli O157, Campylobacter jejuni and Yersinia enterocolitica were used to produce genomic DNA (gDNA). These preparations gave support to method development for qualitative polymerase chain reaction (PCR) detection methods for food-borne pathogens. Purified gDNA was transformed into stable and dry gDNA by using polypropylene vials as carrier and applying a vacuum-drying technique. The gDNA preparations were shown to be sufficiently stable under ambient transport conditions without cooling and proved to have long-term stability at 5degreesC of at least 22 months. The dried DNA was easily reconstituted by addition of distilled water then gentle shaking. These studies have shown that production of stable and dry bacterial gDNA material is feasible and could help satisfy the increasing need for certified reference DNA positive control samples in the field of PCR testing for detection and verification of food-borne microbial pathogens.
Original languageEnglish
JournalAccreditation and Quality Assurance
Volume9
Issue number11-12
Pages (from-to)695-699
ISSN0949-1775
Publication statusPublished - 2004

Keywords

  • standards
  • certified reference material
  • detection pathogenic bacteria
  • genomic DNA
  • PCR

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