TITLE

Determination of viable Escherichia coli using antibody-coated paramagnetic beads with fluorescence detection

AUTHOR(S)
Dudak, F. Ceyda; Boyacı, İsmail H.; Jurkevica, Agnese; Hossain, Mahmud; Aquilar, Zoraida; Halsall, H. Brian; Seliskar, Carl J.; Heineman, William R.
PUB. DATE
February 2009
SOURCE
Analytical & Bioanalytical Chemistry;Feb2009, Vol. 393 Issue 3, p949
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
A rapid and convenient assay system was developed to detect viable Escherichia coli in water. The target bacteria were recovered from solution by immunomagnetic separation and incubated in tryptic soy broth with isopropyl-β- d-thiogalactopyranoside, which induces formation of β-galactosidase in viable bacteria. Lysozyme was used to lyse E. coli cells and release the β-galactosidase. β-Galactosidase converted 4-methylumbelliferyl-β- d-galactoside to 4-methylumbelliferone (4-MU), which was measured by fluorescence spectrophotometry using excitation and emission wavelengths of 355 and 460 nm, respectively. Calibration graphs of 4-MU fluorescence intensity versus E. coli concentration showed a detection range between 8 × 104 and 1.6 × 107 cfu mL−1, with a total analysis time of less than 3 h. The advantage of this method is that it detects viable cells because it is based on the activity of the enzyme intrinsic to live E. coli.
ACCESSION #
36280142

 

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