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Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7.
J Immunol Methods. 2004 Oct; 293(1-2):97-106.JI

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.

Authors+Show Affiliations

United States Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA. agehring@errc.ars.usda.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

15541280

Citation

Gehring, Andrew G., et al. "Enzyme-linked Immunomagnetic Chemiluminescent Detection of Escherichia Coli O157:H7." Journal of Immunological Methods, vol. 293, no. 1-2, 2004, pp. 97-106.
Gehring AG, Irwin PL, Reed SA, et al. Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7. J Immunol Methods. 2004;293(1-2):97-106.
Gehring, A. G., Irwin, P. L., Reed, S. A., Tu, S. I., Andreotti, P. E., Akhavan-Tafti, H., & Handley, R. S. (2004). Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7. Journal of Immunological Methods, 293(1-2), 97-106.
Gehring AG, et al. Enzyme-linked Immunomagnetic Chemiluminescent Detection of Escherichia Coli O157:H7. J Immunol Methods. 2004;293(1-2):97-106. PubMed PMID: 15541280.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7. AU - Gehring,Andrew G, AU - Irwin,Peter L, AU - Reed,Sue A, AU - Tu,Shu-I, AU - Andreotti,Peter E, AU - Akhavan-Tafti,Hashem, AU - Handley,Richard S, PY - 2004/03/31/received PY - 2004/07/06/revised PY - 2004/07/14/accepted PY - 2004/11/16/pubmed PY - 2004/12/16/medline PY - 2004/11/16/entrez SP - 97 EP - 106 JF - Journal of immunological methods JO - J Immunol Methods VL - 293 IS - 1-2 N2 - 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. SN - 0022-1759 UR - https://www.unboundmedicine.com/medline/citation/15541280/Enzyme_linked_immunomagnetic_chemiluminescent_detection_of_Escherichia_coli_O157:H7_ DB - PRIME DP - Unbound Medicine ER -