Development of a flow injection analysis (FIA) immunosensor for the detection of Escherichia coli

Int J Food Microbiol. 1995 Oct;27(2-3):129-37. doi: 10.1016/0168-1605(94)00157-2.

Abstract

A flow injection immunoanalysis (FIA) system has been developed for the detection of Escherichia coli in artificially contaminated food samples. Anti-E. coli antibodies were covalently immobilized onto porous aminopropyl glass beads via glutaraldehyde activation to form an immunoreactor. After adsorption of the cells onto anti-E. coli antibody bound glass beads, 4-methylumbelliferyl-beta-D-glucuronide was injected into the system which was then hydrolyzed by the adsorbed E. coli cells containing beta-D-glucuronidase, an enzyme which is very specific to E. coli and to a few other strains of Shigella. Fluorescent 4-methylumbelliferone released from the enzymatic reaction was then detected by a fluorometer. Owing to the specificity of the antibody towards E. coli, the FIA system was very selective for detection of E. coli whereas Shigella boydii, another GUD-positive bacterium, did not give any response. The FIA system was successfully used for detecting as low as 5 x 10(7) CFU/ml E. coli in less than 30 min and was reusable for at least 300 repeated assays. The immunoreactor yielded reproducible results during 3 months of experimentation if stored overnight at 4 degrees C in carrier buffer containing 0.05 to 0.25% Tween 20.

MeSH terms

  • Escherichia coli / enzymology
  • Escherichia coli / immunology
  • Escherichia coli / isolation & purification*
  • Flow Injection Analysis / instrumentation*
  • Food Microbiology
  • Glucuronosyltransferase / metabolism
  • Reproducibility of Results

Substances

  • 4-methylumbelliferone glucuronyltransferase
  • Glucuronosyltransferase