LUMINESCENT METHODS TO DETECT VIABLE AND TOTAL ESCHERICHIA COLI O157:H7 IN GROUND BEEF+

Autor:	Joe Uknalis, Shiro Yamashoji, Andrew G. Gehring, Shu-I Tu, Peter L. Irwin
Rok vydání:	2005
Předmět:	Biology biology.organism_classification medicine.disease_cause Microbiology Molecular biology chemistry.chemical_compound chemistry Menadione Biochemistry Reagent medicine biology.protein NAD+ kinase Luminescence Escherichia coli Nicotinamide adenine dinucleotide phosphate Bacteria Peroxidase
Zdroj:	Journal of Rapid Methods and Automation in Microbiology. 13:57-70
ISSN:	1745-4581 1060-3999
DOI:	10.1111/j.1745-4581.2005.00010.x
Popis:	Escherichia coli O157:H7 was inoculated into ground beef. Samples were incubated in E. coli broth plus novobiocin growth media at 37C for 5 h. Bacteria were captured by magnetic beads coated with anti-E. coli O157:H7 antibodies and were treated with menadione, a membrane-permeable electron-transfer shuttering reagent, to oxidize the cellular nicotinamide adenine dinucleotide phosphate (NAD[P]H). Alternatively, bead-captured bacteria were sandwiched by second anti-E. coli O157:H7 antibodies labeled with peroxidase. The level of NAD(P)H and the enzyme activity of antibody-bound peroxidase were then measured by luminol-linked luminescence. The treatment of the E. coli O157:H7 with a bacterial protein extraction reagent diminished the luminescence associated with cellular NAD(P)H but not the luminescence associated with the sandwiched peroxidase label. Thus, the NAD(P)H-linked luminescence, not the luminescence that was associated with the sandwiched complexes, was related to the viability of the E. coli O157:H7.
Databáze:	OpenAIRE
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