Evaluation of different DNA extraction procedures for the detection of Salmonella from chicken products by polymerase chain reaction

Lett Appl Microbiol. 1994 Nov;19(5):294-8. doi: 10.1111/j.1472-765x.1994.tb00458.x.

Abstract

Polymerase chain reaction (PCR) was used after a short pre-enrichment culture to detect Salmonella subspecies in chicken fillets. A direct PCR assay performed with chicken meat inoculated with Salmonella Typhimurium produced no PCR products. Six different DNA extraction protocols were tested to recover efficiently Salmonella DNA after a short incubation period. Three of them gave results that were reliable, rapid and sensitive. Successful protocols used Proteinase K and/or a centrifugation step to concentrate the samples. For reliable detection of Salmonella subspecies, a few thousand bacterial cells per ml must be present. To obtain this number of bacterial cells with an inoculation of about one cell in 25 g of ionized food products, it was necessary to incubate samples for at least 10 h before PCR. A larger inoculum of approximately 10 cells in 25 g of ionized food products, required 8 h in culture broth to give positive results by PCR-based assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chickens
  • DNA, Bacterial / isolation & purification*
  • Meat / microbiology*
  • Polymerase Chain Reaction
  • Salmonella / isolation & purification*
  • Sensitivity and Specificity

Substances

  • DNA, Bacterial