Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Curr Microbiol
2011 Feb 01;622:402-8. doi: 10.1007/s00284-010-9721-3.
Show Gene links
Show Anatomy links
Optimization of two immunofluorescent antibodies for the detection of Escherichia coli using immunofluorescent microscopy and flow cytometry.
McCarthy M
,
Culloty SC
.
???displayArticle.abstract???
Two commercially available fluorescein isothiocyanate (FITC) -conjugated anti-Escherichia coli antibodies, tested for immunofluorescence were assessed for their suitability in screening E. coli using flow cytometry. Staining efficacy was initially tested using immunofluorescent microscopy; and further optimization was carried out using flow cytometry. Initially, an acetone fixation step was utilized; however, it was determined statistically that the step could be omitted without impacting the assay and thus reduce the time involved. There was no statistical difference between the staining proficiency of the two antibodies employed. The percentage staining was quite low, approximately 10% for the two antibodies, which indicated that both were equally sensitive but ultimately, more specific antibodies are required for the detection of E. coli. Known proportions of target-E. coli (10⁵, 10⁶, and 10⁷ cells/ml) were mixed with large quantities of non-target bacteria; there was a significant correlation among all the antibodies at the different bacterial cell concentrations. Therefore, despite the low staining percentage achieved on the bacterial cultures, there is a representative and comparative level of staining occurring, between samples and between bacterial strains.
DeCory,
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.
2005, Pubmed
DeCory,
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.
2005,
Pubmed
Deere,
Evaluation of the suitability of bis-(1,3-dibutylbarbituric acid) trimethine oxonol, (diBA-C4(3)-), for the flow cytometric assessment of bacterial viability.
1995,
Pubmed
Gerdts,
FISH and chips: marine bacterial communities analyzed by flow cytometry based on microfluidics.
2006,
Pubmed
Jepras,
Development of a robust flow cytometric assay for determining numbers of viable bacteria.
1995,
Pubmed
Joux,
Use of fluorescent probes to assess physiological functions of bacteria at single-cell level.
2000,
Pubmed
Koch,
Biomass growth rate during the prokaryote cell cycle.
1993,
Pubmed
LaGier,
An electrochemical RNA hybridization assay for detection of the fecal indicator bacterium Escherichia coli.
2005,
Pubmed
Lisle,
Comparison of fluorescence microscopy and solid-phase cytometry methods for counting bacteria in water.
2004,
Pubmed
López-Amorós,
Assessment of E. coli and Salmonella viability and starvation by confocal laser microscopy and flow cytometry using rhodamine 123, DiBAC4(3), propidium iodide, and CTC.
1997,
Pubmed
Makinoshima,
Growth phase-coupled alterations in cell structure and function of Escherichia coli.
2003,
Pubmed
Marie,
Application of the novel nucleic acid dyes YOYO-1, YO-PRO-1, and PicoGreen for flow cytometric analysis of marine prokaryotes.
1996,
Pubmed
Porter,
Fluorescent probes and flow cytometry: new insights into environmental bacteriology.
1996,
Pubmed
Porter,
Rapid, automated separation of specific bacteria from lake water and sewage by flow cytometry and cell sorting.
1993,
Pubmed
Rompré,
Detection and enumeration of coliforms in drinking water: current methods and emerging approaches.
2002,
Pubmed
Roszak,
Survival strategies of bacteria in the natural environment.
1987,
Pubmed
Roth,
Bacterial viability and antibiotic susceptibility testing with SYTOX green nucleic acid stain.
1997,
Pubmed
Tanaka,
Viability of Escherichia coli O157:H7 in natural river water determined by the use of flow cytometry.
2000,
Pubmed
Tortorello,
Quantitative analysis and isolation of Escherichia coli O157:H7 in a food matrix using flow cytometry and cell sorting.
1997,
Pubmed
Veal,
Fluorescence staining and flow cytometry for monitoring microbial cells.
2000,
Pubmed
