Title

Rapid Detection of E. coli Using Flow Cytometry Immunofluorescence

Presenter Information

Clinton Elg

Document Type

Oral Presentation

Location

SURC Ballroom B/C/D

Start Date

21-5-2015

End Date

21-5-2015

Keywords

Escherichia coli, Water Quality, Public Health

Abstract

The bacteria Escherichia coli O157:H7 poses a public health threat as demonstrated in a Washington State lake-associated swimming outbreak that hospitalized eight children in 1999 and the recent outbreak on Mercer Island that closed schools and restaurants for nearly one week as residents boiled drinking water due to contamination. Current methodology of testing for pathogenic bacteria in surface water relies on counting all fecal coliform bacteria with no specificity for pathogenic strains, which are rarely enumerated as the typical laboratory requires three days to positively identify pathogenic E. coli O157:H7. I am developing a flow cytometer assay for rapid (<30 min) detection of E. coli O157:H7 from natural waterways (i.e., streams, lakes, ponds) at detection levels suitable for application to public safety. To date, I have used fluorescent tagged E. coli O157:H7 antibodies (immunofluorescence) to detect the pathogen via flow cytometry in heterogeneous natural stream samples in less than 20 minutes and am incorporating use of a second fluorescent dye to differentiate organic matter from nonorganic debris in mixed water samples to lower the E. coli O157:H7 detection threshold. The ability of this novel flow cytometry assay to speed detection of E. coli O157:H7 from days to minutes holds the potential to protect public health in recreational waters. It also has potential application for use in rapid diagnoses of E. coli O157:H7 in human feces, protection of drinking water quality, and the prevention of food borne illness.

Poster Number

38

Faculty Mentor(s)

Clay Arango, Blaise Dondji

Department/Program

Biological Sciences

Additional Mentoring Department

Biological Sciences

Additional Mentoring Department

Biology

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May 21st, 8:30 AM May 21st, 11:00 AM

Rapid Detection of E. coli Using Flow Cytometry Immunofluorescence

SURC Ballroom B/C/D

The bacteria Escherichia coli O157:H7 poses a public health threat as demonstrated in a Washington State lake-associated swimming outbreak that hospitalized eight children in 1999 and the recent outbreak on Mercer Island that closed schools and restaurants for nearly one week as residents boiled drinking water due to contamination. Current methodology of testing for pathogenic bacteria in surface water relies on counting all fecal coliform bacteria with no specificity for pathogenic strains, which are rarely enumerated as the typical laboratory requires three days to positively identify pathogenic E. coli O157:H7. I am developing a flow cytometer assay for rapid (<30 >min) detection of E. coli O157:H7 from natural waterways (i.e., streams, lakes, ponds) at detection levels suitable for application to public safety. To date, I have used fluorescent tagged E. coli O157:H7 antibodies (immunofluorescence) to detect the pathogen via flow cytometry in heterogeneous natural stream samples in less than 20 minutes and am incorporating use of a second fluorescent dye to differentiate organic matter from nonorganic debris in mixed water samples to lower the E. coli O157:H7 detection threshold. The ability of this novel flow cytometry assay to speed detection of E. coli O157:H7 from days to minutes holds the potential to protect public health in recreational waters. It also has potential application for use in rapid diagnoses of E. coli O157:H7 in human feces, protection of drinking water quality, and the prevention of food borne illness.