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Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method.
Appl Environ Microbiol. 2005 Apr; 71(4):1856-64.AE

Abstract

The objective of this study was to develop and optimize a protocol for the rapid detection of Escherichia coli O157:H7 in aqueous samples by a combined immunomagnetic bead-immunoliposome (IMB/IL) fluorescence assay. The protocol consisted of the filtration or centrifugation of 30- to 100-ml samples followed by incubation of the filter membranes or pellet with anti-E. coli O157:H7 immunomagnetic beads in growth medium specific for E. coli O157:H7. The resulting E. coli O157:H7-immunomagnetic bead complexes were isolated by magnetic separation, washed, and incubated with sulforhodamine B-containing immunoliposomes specific for E. coli O157:H7; the final immunomagnetic bead-E. coli O157:H7-immunoliposome complexes were again isolated by magnetic separation, washed, and lysed with a n-octyl-beta-d-glucopyranoside to release sulforhodamine B. The final protocol took less than 8 h to complete and had a detection limit of less than 1 CFU of E. coli O157:H7 per ml in various aqueous matrices, including apple juice and cider. To validate the protocol at an independent facility, 100-ml samples of groundwater with and without E. coli O157:H7 (15 CFU) were analyzed by a public health laboratory using the optimized protocol and a standard microbiological method. While the IMB/IL fluorescence assay was able to identify E. coli O157:H7-containing samples with 100% accuracy, the standard microbiological method was unable to distinguish E. coli O157:H7-spiked samples from negative controls without further extensive workup. These results demonstrate the feasibility of using immunomagnetic beads in combination with sulforhodamine B-encapsulating immunoliposomes for the rapid detection of E. coli O157:H7 in aqueous samples.

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Authors+Show Affiliations

DeCory TR
Department of Food Science and Technology, Cornell University, Geneva.
Durst RA
No affiliation info available
Zimmerman SJ
No affiliation info available
Garringer LA
No affiliation info available
Paluca G
No affiliation info available
DeCory HH
No affiliation info available
Montagna RA
No affiliation info available

MeSH

BeveragesColony Count, MicrobialEscherichia coli O157FiltrationFluorescenceFresh WaterImmunoassayImmunomagnetic SeparationLiposomesMalusMicropore FiltersRhodaminesTime Factors

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15812012

Citation

DeCory, Thomas R., et al. "Development of an Immunomagnetic Bead-immunoliposome Fluorescence Assay for Rapid Detection of Escherichia Coli O157:H7 in Aqueous Samples and Comparison of the Assay With a Standard Microbiological Method." Applied and Environmental Microbiology, vol. 71, no. 4, 2005, pp. 1856-64.
DeCory TR, Durst RA, Zimmerman SJ, et al. Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method. Appl Environ Microbiol. 2005;71(4):1856-64.
DeCory, T. R., Durst, R. A., Zimmerman, S. J., Garringer, L. A., Paluca, G., DeCory, H. H., & Montagna, R. A. (2005). Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method. Applied and Environmental Microbiology, 71(4), 1856-64.
DeCory TR, et al. Development of an Immunomagnetic Bead-immunoliposome Fluorescence Assay for Rapid Detection of Escherichia Coli O157:H7 in Aqueous Samples and Comparison of the Assay With a Standard Microbiological Method. Appl Environ Microbiol. 2005;71(4):1856-64. PubMed PMID: 15812012.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method. AU - DeCory,Thomas R, AU - Durst,Richard A, AU - Zimmerman,Scott J, AU - Garringer,Linda A, AU - Paluca,Gary, AU - DeCory,Heleen H, AU - Montagna,Richard A, PY - 2005/4/7/pubmed PY - 2005/7/7/medline PY - 2005/4/7/entrez SP - 1856 EP - 64 JF - Applied and environmental microbiology JO - Appl Environ Microbiol VL - 71 IS - 4 N2 - The objective of this study was to develop and optimize a protocol for the rapid detection of Escherichia coli O157:H7 in aqueous samples by a combined immunomagnetic bead-immunoliposome (IMB/IL) fluorescence assay. The protocol consisted of the filtration or centrifugation of 30- to 100-ml samples followed by incubation of the filter membranes or pellet with anti-E. coli O157:H7 immunomagnetic beads in growth medium specific for E. coli O157:H7. The resulting E. coli O157:H7-immunomagnetic bead complexes were isolated by magnetic separation, washed, and incubated with sulforhodamine B-containing immunoliposomes specific for E. coli O157:H7; the final immunomagnetic bead-E. coli O157:H7-immunoliposome complexes were again isolated by magnetic separation, washed, and lysed with a n-octyl-beta-d-glucopyranoside to release sulforhodamine B. The final protocol took less than 8 h to complete and had a detection limit of less than 1 CFU of E. coli O157:H7 per ml in various aqueous matrices, including apple juice and cider. To validate the protocol at an independent facility, 100-ml samples of groundwater with and without E. coli O157:H7 (15 CFU) were analyzed by a public health laboratory using the optimized protocol and a standard microbiological method. While the IMB/IL fluorescence assay was able to identify E. coli O157:H7-containing samples with 100% accuracy, the standard microbiological method was unable to distinguish E. coli O157:H7-spiked samples from negative controls without further extensive workup. These results demonstrate the feasibility of using immunomagnetic beads in combination with sulforhodamine B-encapsulating immunoliposomes for the rapid detection of E. coli O157:H7 in aqueous samples. SN - 0099-2240 UR - https://www.unboundmedicine.com/medline/citation/15812012/Development_of_an_immunomagnetic_bead_immunoliposome_fluorescence_assay_for_rapid_detection_of_Escherichia_coli_O157:H7_in_aqueous_samples_and_comparison_of_the_assay_with_a_standard_microbiological_method_ DB - PRIME DP - Unbound Medicine ER -