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Comparison of enzyme-linked immunomagnetic chemiluminescence with U.S. Food and Drug Administration's Bacteriological Analytical Manual method for the detection of Escherichia coli O157:H7.
J Microbiol Methods. 2006 Dec; 67(3):527-33.JM

Abstract

Escherichia coli O157:H7, a major foodborne pathogen, has been associated with numerous cases of foodborne illnesses. Rapid methods have been developed for the screening of this pathogen in foods in order to circumvent timely plate culture techniques. Unfortunately, many rapid methods are presumptive and do not claim to confirm the presence of E. coli O157:H7. The previously developed method, enzyme-linked immunomagnetic chemiluminescence (ELIMCL), has been improved upon to allow for fewer incidences of false positives when used to detect E. coli O157:H7 in the presence of mixed cultures. The key feature of this assay is that it combines the highly selective synergism of both anti-O157 and anti-H7 antibodies in the sandwich immunoassay format. This work presents application of a newly semi-automated version of ELIMCL to the detection of E. coli O157:H7 in pristine buffered saline yielding detection limits of approximately 1 x 10(5) to 1 x 10(6) of live cells/mL. ELIMCL was further demonstrated to detect E. coli O157:H7 inoculated into artificially contaminated ground beef at ca. 400 CFU/g after a 5 h enrichment and about 1.5 h assay time for a total detection time of about 6.5 h. Finally, ELIMCL was compared with USFDA's Bacteriological Analytical Manual method for E. coli O157:H7 in a double-blind study. Using McNemar's treatment, the two methods were determined to be statistically similar for the detection of E. coli O157:H7 in ground beef inoculated with mixed cultures of select bacteria.

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Authors+Show Affiliations

Gehring 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 <agehring@errc.ars.usda.gov>
Albin DM
No affiliation info available
Irwin PL
No affiliation info available
Reed SA
No affiliation info available
Tu SI
No affiliation info available

MeSH

Bacteriological TechniquesEscherichia coli O157Food MicrobiologyImmunomagnetic SeparationLuminescenceMeatSensitivity and SpecificityUnited StatesUnited States Food and Drug Administration

Pub Type(s)

Comparative Study
Journal Article
Validation Study

Language

eng

PubMed ID

16935371

Citation

Gehring, Andrew G., et al. "Comparison of Enzyme-linked Immunomagnetic Chemiluminescence With U.S. Food and Drug Administration's Bacteriological Analytical Manual Method for the Detection of Escherichia Coli O157:H7." Journal of Microbiological Methods, vol. 67, no. 3, 2006, pp. 527-33.
Gehring AG, Albin DM, Irwin PL, et al. Comparison of enzyme-linked immunomagnetic chemiluminescence with U.S. Food and Drug Administration's Bacteriological Analytical Manual method for the detection of Escherichia coli O157:H7. J Microbiol Methods. 2006;67(3):527-33.
Gehring, A. G., Albin, D. M., Irwin, P. L., Reed, S. A., & Tu, S. I. (2006). Comparison of enzyme-linked immunomagnetic chemiluminescence with U.S. Food and Drug Administration's Bacteriological Analytical Manual method for the detection of Escherichia coli O157:H7. Journal of Microbiological Methods, 67(3), 527-33.
Gehring AG, et al. Comparison of Enzyme-linked Immunomagnetic Chemiluminescence With U.S. Food and Drug Administration's Bacteriological Analytical Manual Method for the Detection of Escherichia Coli O157:H7. J Microbiol Methods. 2006;67(3):527-33. PubMed PMID: 16935371.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparison of enzyme-linked immunomagnetic chemiluminescence with U.S. Food and Drug Administration's Bacteriological Analytical Manual method for the detection of Escherichia coli O157:H7. AU - Gehring,Andrew G, AU - Albin,David M, AU - Irwin,Peter L, AU - Reed,Sue A, AU - Tu,Shu-I, Y1 - 2006/08/28/ PY - 2006/01/01/received PY - 2006/04/19/revised PY - 2006/04/19/accepted PY - 2006/8/29/pubmed PY - 2007/2/13/medline PY - 2006/8/29/entrez SP - 527 EP - 33 JF - Journal of microbiological methods JO - J Microbiol Methods VL - 67 IS - 3 N2 - Escherichia coli O157:H7, a major foodborne pathogen, has been associated with numerous cases of foodborne illnesses. Rapid methods have been developed for the screening of this pathogen in foods in order to circumvent timely plate culture techniques. Unfortunately, many rapid methods are presumptive and do not claim to confirm the presence of E. coli O157:H7. The previously developed method, enzyme-linked immunomagnetic chemiluminescence (ELIMCL), has been improved upon to allow for fewer incidences of false positives when used to detect E. coli O157:H7 in the presence of mixed cultures. The key feature of this assay is that it combines the highly selective synergism of both anti-O157 and anti-H7 antibodies in the sandwich immunoassay format. This work presents application of a newly semi-automated version of ELIMCL to the detection of E. coli O157:H7 in pristine buffered saline yielding detection limits of approximately 1 x 10(5) to 1 x 10(6) of live cells/mL. ELIMCL was further demonstrated to detect E. coli O157:H7 inoculated into artificially contaminated ground beef at ca. 400 CFU/g after a 5 h enrichment and about 1.5 h assay time for a total detection time of about 6.5 h. Finally, ELIMCL was compared with USFDA's Bacteriological Analytical Manual method for E. coli O157:H7 in a double-blind study. Using McNemar's treatment, the two methods were determined to be statistically similar for the detection of E. coli O157:H7 in ground beef inoculated with mixed cultures of select bacteria. SN - 0167-7012 UR - https://www.unboundmedicine.com/medline/citation/16935371/Comparison_of_enzyme_linked_immunomagnetic_chemiluminescence_with_U_S__Food_and_Drug_Administration's_Bacteriological_Analytical_Manual_method_for_the_detection_of_Escherichia_coli_O157:H7_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0167-7012(06)00157-6 DB - PRIME DP - Unbound Medicine ER -