Rapid enumeration of Escherichia coli in oysters by a quantitative PCR-ELISA

J Appl Microbiol. 1999 Feb;86(2):231-6. doi: 10.1046/j.1365-2672.1999.00659.x.

Abstract

Direct enumeration of Escherichia coli from oysters was achieved using a polymerase chain reaction (PCR) amplification of the lamB gene coupled with an enzyme-linked immunosorbent assay (ELISA). Amplified PCR products generated using a digoxigenin-labelled primer were heat denatured before being quantified by an ELISA. A biotinylated probe immobilized onto streptavidin-coated microplates was used to capture the digoxigenin-labelled fragments that were detected with a peroxidase antidigoxigenin conjugate. Subsequent enzymic conversion of substrate gave distinct absorbance differences when assaying oyster samples containing E. coli in the range 10-10(5) cfu g-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Outer Membrane Proteins
  • Colony Count, Microbial
  • DNA Primers
  • Digoxigenin
  • Enzyme-Linked Immunosorbent Assay*
  • Escherichia coli / genetics
  • Escherichia coli / isolation & purification*
  • Ostreidae / microbiology*
  • Polymerase Chain Reaction / methods*
  • Porins
  • Receptors, Virus / genetics
  • Sensitivity and Specificity

Substances

  • Bacterial Outer Membrane Proteins
  • DNA Primers
  • Porins
  • Receptors, Virus
  • maltoporins
  • Digoxigenin