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

21 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

Fingerprint

enterotoxins
Milk
milk
flow cytometry
immunoglobulin G
Flow Cytometry
Immunomagnetic Separation
immunomagnetic separation
Immunoglobulin G
Fluorescence
fluorescence
antibodies
Antibodies
rabbits
Rabbits
staphylococcal enterotoxin B
Particle Size
Goats
sampling
Limit of Detection

All Science Journal Classification (ASJC) codes

  • Food Science
  • Microbiology

Cite this

Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk. / Miyamoto, Takahisa; Kamikado, Hideaki; Kobayashi, Hiroshi; Honjoh, Ken-ichi; Iio, Masayoshi.

In: Journal of Food Protection, Vol. 66, No. 7, 01.07.2003, p. 1222-1226.

Research output: Contribution to journalArticle

Miyamoto, Takahisa ; Kamikado, Hideaki ; Kobayashi, Hiroshi ; Honjoh, Ken-ichi ; Iio, Masayoshi. / Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk. In: Journal of Food Protection. 2003 ; Vol. 66, No. 7. pp. 1222-1226.
@article{a5a49cdf4cda40dcadcbc5ce3d409b3c,
title = "Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk",
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.",
author = "Takahisa Miyamoto and Hideaki Kamikado and Hiroshi Kobayashi and Ken-ichi Honjoh and Masayoshi Iio",
year = "2003",
month = "7",
day = "1",
doi = "10.4315/0362-028X-66.7.1222",
language = "English",
volume = "66",
pages = "1222--1226",
journal = "Journal of Food Protection",
issn = "0362-028X",
publisher = "International Association for Food Protection",
number = "7",

}

TY - JOUR

T1 - Immunomagnetic flow cytometric detection of staphylococcal enterotoxin B in raw and dry milk

AU - Miyamoto, Takahisa

AU - Kamikado, Hideaki

AU - Kobayashi, Hiroshi

AU - Honjoh, Ken-ichi

AU - Iio, Masayoshi

PY - 2003/7/1

Y1 - 2003/7/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0037770001&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037770001&partnerID=8YFLogxK

U2 - 10.4315/0362-028X-66.7.1222

DO - 10.4315/0362-028X-66.7.1222

M3 - Article

C2 - 12870756

AN - SCOPUS:0037770001

VL - 66

SP - 1222

EP - 1226

JO - Journal of Food Protection

JF - Journal of Food Protection

SN - 0362-028X

IS - 7

ER -