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Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7

Authors :
Peter L. Irwin
Sue Reed
Andrew G. Gehring
Shu-I Tu
Hashem Akhavan-Tafti
Peter E. Andreotti
Richard S. Handley
Source :
Journal of Immunological Methods. 293:97-106
Publication Year :
2004
Publisher :
Elsevier BV, 2004.

Abstract

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.

Details

ISSN :
00221759
Volume :
293
Database :
OpenAIRE
Journal :
Journal of Immunological Methods
Accession number :
edsair.doi.dedup.....f04778669e9f5aa30407074f3ad41332
Full Text :
https://doi.org/10.1016/j.jim.2004.07.004