title: Bacteriophage-based biosorbents coupled with bioluminescent ATP assay for rapid concentration and detection of Escherichia coli.
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.
authors.affiliation: Canadian Research Institute for Food Safety, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
authors.forename: O
authors.initials: O
authors.lastname: Minikh
authors.forename: M
authors.initials: M
authors.lastname: Tolba
authors.forename: L Y
authors.initials: LY
authors.lastname: Brovko
authors.forename: M W
authors.initials: MW
authors.lastname: Griffiths
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D001431:Bacteriological Techniques; D001433:Bacteriolysis; D017122:Bacteriophage T4 / Q000235:genetics / Q000502:physiology*; D001710:Biotin / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry / Q000302:isolation & purification* / Q000821:virology; D005374:Filtration / Q000379:methods*; D049449:Luminescence; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D053585:Virus Attachment; D014871:Water Microbiology
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D001710:Biotin; D000255:Adenosine Triphosphate; D002482:Cellulose