| Novel immuno-FRET assay method for Bacillus spores and Escherichia coli O157:H7. | |
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MedLine Citation:
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PMID: 11573946 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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J G Bruno; S J Ulvick; G L Uzzell; J S Tabb; E R Valdes; C A Batt |
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Publication Detail:
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Type: Evaluation Studies; Journal Article; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Biochemical and biophysical research communications Volume: 287 ISSN: 0006-291X ISO Abbreviation: Biochem. Biophys. Res. Commun. Publication Date: 2001 Oct |
Date Detail:
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Created Date: 2001-09-27 Completed Date: 2001-12-04 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0372516 Medline TA: Biochem Biophys Res Commun Country: United States |
Other Details:
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Languages: eng Pagination: 875-80 Citation Subset: IM |
Copyright Information:
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Copyright 2001 Academic Press. |
Affiliation:
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OmniSite BioDiagnostics, Incorporated, 101 West Sixth Street, Suite 200, Austin, TX 78701-2932, USA. bruno@spec.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Bacillus cereus
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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:
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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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