Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six coccidiostats in feed and eggs

M.E. Bienenmann-Ploum, A.C. Huet, K. Campbell, T.L. Fodey, U. Vincent, P. Delahaut, C.T. Elliot, M.W.F. Nielen

Research output: Contribution to journalArticleAcademicpeer-review

18 Citations (Scopus)

Abstract

Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority coccidiostats. Different coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4'-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 µg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 µg/kg, respectively.
Original languageEnglish
Pages (from-to)1361-1373
JournalAnalytical and Bioanalytical Chemistry
Volume404
Issue number5
DOIs
Publication statusPublished - 2012

Fingerprint

Coccidiostats
Immunoassay
Eggs
Screening
diclazuril
European Union
Nicarbazin
Lasalocid
Veterinary Drugs
Coccidiosis
Monensin
Poultry
Flow cytometry
Microspheres
Limit of Detection
Immune Sera
Assays
Carrier Proteins
Flow Cytometry
Color

Keywords

  • time-resolved fluoroimmunoassay
  • veterinary drug residues
  • additive nicarbazin
  • immunosorbent-assay
  • tissue distribution
  • poultry
  • salinomycin
  • chickens
  • contamination
  • validation

Cite this

Bienenmann-Ploum, M.E. ; Huet, A.C. ; Campbell, K. ; Fodey, T.L. ; Vincent, U. ; Delahaut, P. ; Elliot, C.T. ; Nielen, M.W.F. / Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six coccidiostats in feed and eggs. In: Analytical and Bioanalytical Chemistry. 2012 ; Vol. 404, No. 5. pp. 1361-1373.
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abstract = "Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority coccidiostats. Different coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4'-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 µg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 µg/kg, respectively.",
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six coccidiostats in feed and eggs. / Bienenmann-Ploum, M.E.; Huet, A.C.; Campbell, K.; Fodey, T.L.; Vincent, U.; Delahaut, P.; Elliot, C.T.; Nielen, M.W.F.

In: Analytical and Bioanalytical Chemistry, Vol. 404, No. 5, 2012, p. 1361-1373.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six coccidiostats in feed and eggs

AU - Bienenmann-Ploum, M.E.

AU - Huet, A.C.

AU - Campbell, K.

AU - Fodey, T.L.

AU - Vincent, U.

AU - Delahaut, P.

AU - Elliot, C.T.

AU - Nielen, M.W.F.

PY - 2012

Y1 - 2012

N2 - Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority coccidiostats. Different coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4'-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 µg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 µg/kg, respectively.

AB - Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority coccidiostats. Different coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4'-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 µg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 µg/kg, respectively.

KW - time-resolved fluoroimmunoassay

KW - veterinary drug residues

KW - additive nicarbazin

KW - immunosorbent-assay

KW - tissue distribution

KW - poultry

KW - salinomycin

KW - chickens

KW - contamination

KW - validation

U2 - 10.1007/s00216-012-6214-1

DO - 10.1007/s00216-012-6214-1

M3 - Article

VL - 404

SP - 1361

EP - 1373

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

IS - 5

ER -