Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk

Takahisa Miyamoto, Hideaki Kamikado, Hiroshi Kobayashi, Ken-ichi Honjoh, Masayoshi Iio

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A rapid and sensitive method for detection of staphylococcal enterotoxin B (SEB) in raw and dry milk samples with the use of antibody-based immunomagnetic separation (IMS) in conjunction with flow cytometry (FCM) was developed. Sheep anti-SEB immunoglobulin G (IgG) was immobilized on Dynabeads M-280. The SEB initially binds to the capturing antibody, which is bound on the magnetic beads. The rabbit anti-SEB IgG binds to the captured toxin and is further labeled with a Cy5-labeled goat anti-rabbit IgG antibody. The percentage of the beads that were fluorescent was measured by FCM. FCM was carried out for 1 min, and the data obtained were expressed as histograms for particle size (forward light scatter) and histograms for fluorescence intensity. A peak corresponding to the magnetic beads was clearly distinguished from a peak derived from contaminating particles in the sample solution. In the absence of SEB, about 10% of the beads emitted fluorescence. The percentage of fluorescent beads and the fluorescence intensity increased with increasing SEB concentrations. For this IMS-FCM assay, the lower limits of detection for SEB were estimated to be 0.01 and 0.25 ng/ml for buffer and milk samples, respectively.

Original languageEnglish
Pages (from-to)1222-1226
Number of pages5
JournalJournal of Food Protection
Volume66
Issue number7
DOIs
Publication statusPublished - Jul 1 2003

All Science Journal Classification (ASJC) codes

  • Food Science
  • Microbiology

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