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Detection of Escherichia coli O157:H7 in raw meat by immunomagnetic separation and multiplex PCR.
Acta Microbiol Pol. 2002; 51(4):327-37.AM

Abstract

The aim of this research was to elaborate fast and sensitive method ofdetection of E. coli O157:H7 in food samples. Raw ground meat obtained from retail was artificially inoculated with low numbers of E. coli O157:H7. 18 h enrichment culture allowed pathogenic bacteria to multiply to the levels detectable in multiplex PCR. Immunomagnetic separation with magnetic beads coated with an antibody against E. coli O157:H7 were used to concentrate target bacteria and to separate PCR inhibitors. A portion of the bacterial suspension was used in a multiplex PCR to amplify eae (attaching and effacing) gene, stx (shiga toxin) genes and 90 kbp plasmid. The sensitivity of E. coli O157:H7 detection method was shown to be 1 cfu per 25 g of food sample. The total analysis can be completed within 24 h, whilst traditional methods involves enrichment, direct plating and confirmation tests with entire time at least 3 days.

Authors+Show Affiliations

Lekowska-Kochaniak A
Department of Microbiology, National Food and Nutrition Institute, Powsińska 61/63, 02-903 Warsaw, Poland.
Czajkowska D
No affiliation info available
Popowski J
No affiliation info available

MeSH

AnimalsCattleEscherichia coli O157Immunomagnetic SeparationMeatPolymerase Chain ReactionSensitivity and Specificity

Pub Type(s)

Journal Article

Language

eng

PubMed ID

12708821

Citation

Lekowska-Kochaniak, Aneta, et al. "Detection of Escherichia Coli O157:H7 in Raw Meat By Immunomagnetic Separation and Multiplex PCR." Acta Microbiologica Polonica, vol. 51, no. 4, 2002, pp. 327-37.
Lekowska-Kochaniak A, Czajkowska D, Popowski J. Detection of Escherichia coli O157:H7 in raw meat by immunomagnetic separation and multiplex PCR. Acta Microbiol Pol. 2002;51(4):327-37.
Lekowska-Kochaniak, A., Czajkowska, D., & Popowski, J. (2002). Detection of Escherichia coli O157:H7 in raw meat by immunomagnetic separation and multiplex PCR. Acta Microbiologica Polonica, 51(4), 327-37.
Lekowska-Kochaniak A, Czajkowska D, Popowski J. Detection of Escherichia Coli O157:H7 in Raw Meat By Immunomagnetic Separation and Multiplex PCR. Acta Microbiol Pol. 2002;51(4):327-37. PubMed PMID: 12708821.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of Escherichia coli O157:H7 in raw meat by immunomagnetic separation and multiplex PCR. AU - Lekowska-Kochaniak,Aneta, AU - Czajkowska,Danuta, AU - Popowski,Janusz, PY - 2003/4/24/pubmed PY - 2003/5/16/medline PY - 2003/4/24/entrez SP - 327 EP - 37 JF - Acta microbiologica Polonica JO - Acta Microbiol Pol VL - 51 IS - 4 N2 - The aim of this research was to elaborate fast and sensitive method ofdetection of E. coli O157:H7 in food samples. Raw ground meat obtained from retail was artificially inoculated with low numbers of E. coli O157:H7. 18 h enrichment culture allowed pathogenic bacteria to multiply to the levels detectable in multiplex PCR. Immunomagnetic separation with magnetic beads coated with an antibody against E. coli O157:H7 were used to concentrate target bacteria and to separate PCR inhibitors. A portion of the bacterial suspension was used in a multiplex PCR to amplify eae (attaching and effacing) gene, stx (shiga toxin) genes and 90 kbp plasmid. The sensitivity of E. coli O157:H7 detection method was shown to be 1 cfu per 25 g of food sample. The total analysis can be completed within 24 h, whilst traditional methods involves enrichment, direct plating and confirmation tests with entire time at least 3 days. SN - 0137-1320 UR - https://www.unboundmedicine.com/medline/citation/12708821/Detection_of_Escherichia_coli_O157:H7_in_raw_meat_by_immunomagnetic_separation_and_multiplex_PCR_ DB - PRIME DP - Unbound Medicine ER -