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Anal Chem. 1993 Aug 1;65(15):2036-9.

Detection and removal of Escherichia coli using fluorescein isothiocyanate conjugated monoclonal antibody immobilized on bacterial magnetic particles.

Author information

1
Department of Biotechnology, Tokyo University of Agriculture and Technology, Japan.

Abstract

A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 10(2)-10(6) cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E. coli from the bacterial suspension.

PMID:
8372968
DOI:
10.1021/ac00063a018
[Indexed for MEDLINE]

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