Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7.
E. coli O157:H7 is a pathogenic microorganism that has been implicated in numerous cases of foodborne illnesses. A variety of rapid methods exist that show promise for the presumptive detection of this pathogen without the immediate need for incubating test samples for hours to days in microbial enrichment and culture media. In recent years, highly sensitive chemiluminescence has become a more affordable and portable detection method. Chemiluminescent detection has been coupled with the selectivity of antibodies, magnetic microparticle separation/isolation, and enzymatic signal amplification in order to develop a rapid method, termed enzyme-linked immunomagnetic chemiluminescence (ELIMCL). This work presents the application of ELIMCL to the detection of E. coli O157:H7 in pristine buffered saline with a detection limit of 7.6 x 10(3) for live cells in approx. 75 min assay time. The blocking agent casein and the surfactant Tween 20 were used to lower background luminescence and thus maximize signal-to-noise ratios. After 5.5 h of enrichment culture, ELIMCL was demonstrated to detect E. coli O157:H7 inoculated in ground beef at 10 CFU/g in a total assay time of about 7 h.
Gehring
Andrew G
AG
United States Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA. agehring@errc.ars.usda.gov
Irwin
Peter L
PL
Reed
Sue A
SA
Tu
Shu-I
SI
Andreotti
Peter E
PE
Akhavan-Tafti
Hashem
H
Handley
Richard S
RS
D004797:Enzyme-Linked Immunosorbent Assay; D019453:Escherichia coli O157 / Q000276:immunology / Q000302:isolation & purification*; D018189:Immunomagnetic Separation; D008163:Luminescent Measurements; D013997:Time Factors