Document Detail


Novel immuno-FRET assay method for Bacillus spores and Escherichia coli O157:H7.
MedLine Citation:
PMID:  11573946     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Novel immunofluorescence resonance energy transfer (immuno-FRET) assays for both Bacillus cereus spores and Escherichia coli O157:H7 are reported. Both assays involve the use of dual (QSY-7 and Oregon Green 514-antibody)-labeled spores or vegetative bacteria, such that Oregon Green 514-labeled antibodies are quenched by proximal QSY-7 molecules that are covalently bound to the dual (Oregon Green 514 and QSY-7)-labeled cells. Upon introduction of unlabeled bacteria or spores, in the respective assays, an increase in fluorescence is observed in proportion to the numbers of unlabeled cells. This is due to migration of Oregon Green 514-labeled antibody from the dual-labeled cells to the unlabeled target cells as verified by fluorescence microscopy. Optimization of the QSY-7 surface density led to a B. cereus spore detection sensitivity of approximately 1.0 x 10(5) to 2.5 x 10(5) spores per milliliter and 3.5 x 10(5) cells per milliliter for E. coli using a conventional cuvette-based spectrofluorometer.
Authors:
J G Bruno; S J Ulvick; G L Uzzell; J S Tabb; E R Valdes; C A Batt
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  287     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-09-27     Completed Date:  2001-12-04     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  United States    
Other Details:
Languages:  eng     Pagination:  875-80     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
Affiliation:
OmniSite BioDiagnostics, Incorporated, 101 West Sixth Street, Suite 200, Austin, TX 78701-2932, USA. bruno@spec.com
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MeSH Terms
Descriptor/Qualifier:
Bacillus cereus / isolation & purification*
Energy Transfer
Escherichia coli O157 / isolation & purification*
Fluoresceins / metabolism
Fluorescent Antibody Technique*
Fluorescent Dyes / metabolism
Food Microbiology
Microscopy, Fluorescence
Molecular Structure
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Fluorescence
Spores, Bacterial / isolation & purification*
Chemical
Reg. No./Substance:
0/Fluoresceins; 0/Fluorescent Dyes; 0/oregon green 514

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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