Development of a sensitive chemiluminometric assay for the detection of beta-galactosidase in permeabilized coliform bacteria and comparison with fluorometry and colorimetry

Author:

Van Poucke S O1,Nelis H J1

Affiliation:

1. Department of Pharmaceutical Analysis, University of Ghent, Belgium.

Abstract

We developed a chemiluminometric assay of beta-galactosidase in coliform bacteria, using a phenylgalactose-substituted 1,2-dioxetane derivative as a substrate. Permeabilization of cells is required to ensure the efficient cellular uptake of this compound. By this method, one coliform seeded in 100 ml of sterile water can be detected after a 6- to 9-h propagation phase followed by a 45-min enzyme assay in the presence of polymyxin B. Compared with fluorometry and colorimetry, chemiluminometry afforded 4- and 1,000-fold increases in sensitivity and 1- and 6-h increases in the speed of detection, respectively.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Reference31 articles.

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2. American Public Health Association. 1992. Standard methods for the examination of water and wastewater 18th ed. American Public Health Association Washington D.C.

3. A rapid and simple chemiluminescent assay for Escherichia coli ~-galactosidase;Beale E. G.;BioTechniques,1992

4. Detection of coliform bacteria in water by polymerase chain reaction and gene probes;Bej A. K.;Appl. Environ. Microbiol.,1990

5. Rapid detection of total and fecal coliforms in water by enzymatic hydrolysis of 4-methylumbelliferone-~-D-galactoside;Berg J. D.;Appl. Environ. Microbiol.,1988

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