Bacteriophage-based biosorbents coupled with bioluminescent ATP assay for rapid concentration and detection of Escherichia coli.
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.
Minikh
O
O
Canadian Research Institute for Food Safety, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
Tolba
M
M
Brovko
L Y
LY
Griffiths
M W
MW
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
D011994:Recombinant Proteins; D014764:Viral Proteins; D001710:Biotin; D000255:Adenosine Triphosphate; D002482:Cellulose