Analytical and Bioanalytical Chemistry

, Volume 407, Issue 15, pp 4459–4472 | Cite as

Development of a rapid multiplexed assay for the direct screening of antimicrobial residues in raw milk

  • Terry F. McGrath
  • Laura McClintock
  • John S. Dunn
  • Gregory M. Husar
  • Michael J. Lochhead
  • Ronald W. Sarver
  • Frank E. Klein
  • Jennifer A. Rice
  • Katrina Campbell
  • Christopher T. Elliott
Research Paper
Part of the following topical collections:
  1. Hormone and Veterinary Drug Residue Analysis


Antimicrobial residues found to be present in milk can have both health and economic impacts. For these reasons, the widespread routine testing of milk is required. Due to delays with sample handling and test scheduling, laboratory-based tests are not always suited for making decisions about raw material intake and product release, especially when samples require shipping to a central testing facility. Therefore, rapid on-site screening tests that can produce results within a matter of minutes are required to facilitate rapid intake and product release processes. Such tests must be simple for use by non-technical staff. There is increasing momentum towards the development and implementation of multiplexing tests that can detect a range of important antimicrobial residues simultaneously. A simple in situ multiplexed planar waveguide device that can simultaneously detect chloramphenicol, streptomycin and desfuroylceftiofur in raw dairy milk, without sample preparation, has been developed. Samples are simply mixed with antibody prior to an aliquot being passed through the detection cartridge for 5 min before reading on a field-deployable portable instrument. Multiplexed calibration curves were produced in both buffer and raw milk. Buffer curves, for chloramphenicol, streptomycin and desfuroylceftiofur, showed linear ranges (inhibitory concentration (IC)20–IC80) of 0.1–0.9, 3–129 and 12–26 ng/ml, whilst linear range in milk was 0.13–0.74, 11–376 and 2–12 ng/ml, respectively, thus meeting European legislated concentration requirements for both chloramphenicol and streptomycin, in milk, without the need for any sample preparation. Desfuroylceftiofur-contaminated samples require only simple sample dilution to bring positive samples within the range of quantification. Assay repeatability and reproducibility were lower than 12 coefficient of variation (%CV), whilst blank raw milk samples (n = 9) showed repeatability ranging between 4.2 and 8.1 %CV when measured on all three calibration curves.

Graphical Abstract

MBio SnapEsi reader and cartridge


Rapid multiplexed screening Veterinary drug residue milk No sample preparation Planar waveguide On-site screening 


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Terry F. McGrath
    • 1
  • Laura McClintock
    • 1
  • John S. Dunn
    • 2
  • Gregory M. Husar
    • 2
  • Michael J. Lochhead
    • 2
  • Ronald W. Sarver
    • 3
  • Frank E. Klein
    • 3
  • Jennifer A. Rice
    • 3
  • Katrina Campbell
    • 1
  • Christopher T. Elliott
    • 1
  1. 1.Institute for Global Food Security, School of Biological SciencesQueen’s UniversityBelfastUK
  2. 2.MBio Diagnostics IncBoulderUSA
  3. 3.Neogen CorporationLansingUSA

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