alexa Bacteriophage-based biosorbents coupled with bioluminescent ATP assay for rapid concentration and detection of Escherichia coli.


Biosensors Journal

Author(s): Minikh O, Tolba M, Brovko LY, Griffiths MW, Minikh O, Tolba M, Brovko LY, Griffiths MW

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Abstract Wild type T4 bacteriophage and recombinant T4 bacteriophages displaying biotin binding peptide (BCCP) and cellulose binding module (CBM) on their heads were immobilized on nano-aluminum fiber-based filter (Disruptor), streptavidin magnetic beads and microcrystalline cellulose, respectively. Infectivity of the immobilized phages was investigated by monitoring the phage-mediated growth inhibition of bioluminescent E. coli B and cell lysis using bioluminescent ATP assay. The results showed that phage immobilization resulted in a partial loss of infectivity as compared with the free phage. Nevertheless, the use of a biosorbent based on T4 bacteriophage immobilized on Disruptor filter coupled with a bioluminescent ATP assay allowed simultaneous concentration and detection of as low as 6 x 10(3)cfu/mL of E. coli in the sample within 2h with high accuracy (CV=1-5\% in log scale). Excess of interfering microflora at levels 60-fold greater than the target organism did not affect the results when bacteriophage was immobilized on the filter prior to concentration of bacterial cells. This article was published in J Microbiol Methods and referenced in Biosensors Journal

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