Preparation of milk samples for PCR analysis using a rapid filtration technique

S. J. Wu, C. I. Kado

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Aims: To investigate the usefulness of a straightforward filtration method for the isolation of Escherichia coli O157:H7 contaminants from milk for PCR detection. Methods and Results: Escherichia coli O157:H7 is grown in milk and enriched in Luria-Bertani (LB) medium. Samples are filtered through a 0.45-μm pore membrane. The membrane is immersed in 200-μl lysis buffer and incubated at 95°C for 10 min to release bacterial DNA for subsequent PCR detection. Under current conditions, the overall duration from filtration to PCR-ready DNA generation is <20 min, and the detection level for PCR was as low as 10 CFU of bacteria in 1 ml of milk. Conclusion: Bacterial contaminants of milk can be concentrated and isolated by a simple, one-step filtration and their DNA can be released for subsequent PCR detection by heating the filter membrane at 95°C for 10 min. Significance and Impact of the Study: The simplicity of this method allows inexpensive, high throughput automation that meets the demands of modern food hygiene monitoring.

Original languageEnglish
Pages (from-to)1342-1346
Number of pages5
JournalJournal of Applied Microbiology
Volume96
Issue number6
DOIs
StatePublished - 2004

Keywords

  • Detection
  • Escherichia coli
  • Filtration
  • Food analysis
  • PCR

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