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Abstract

In this study, we show a significantly reduced assay time and a greatly increased bead recovery for a commercial Luminex-based multiplex diagnostic immunoassay by performing all liquid handling steps of the assay protocol in a non-contact acoustic trapping platform. The Luminex assay is designed for detecting antibodies in poultry serum for infectious bursal disease virus, infectious bronchitis virus, Newcastle disease virus and avian reovirus. Here, we show proof-of-concept of a microfluidic system capable of being fully automated and handling samples in a parallel format with a miniature physical footprint where the affinity beads are retained in a non-contact levitated mode in a glass capillary throughout the assay protocol. The different steps are: incubation with the serum sample, secondary antibodies and fluorescent reporters and finally washing to remove any non-specifically bound species. A Luminex 200 instrument was used for the readout. The flow rates applied to the capillary during the initial trapping event and the wash steps were optimised for maximum bead recovery, resulting in a bead recovery of 75% for the complete assay. This can be compared to a bead recovery of approximately 30% when an automatic wash station was used when the assay was performed in the conventional manual format.

Keywords

Acoustic trapping; Immuno-based assay; Multiplex; Luminex

Published in

Analytica Chimica Acta
2015, volume: 853, pages: 682-688

SLU Authors

  • Xia, Hongyan

    • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    • World Organization for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine

  • Belak, Sandor

    • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    • National Veterinary Institute (SVA)
    • World Organization for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine

UKÄ Subject classification

Medical Bioscience

Publication identifier

  • DOI: https://doi.org/10.1016/j.aca.2014.07.008

Permanent link to this page (URI)

https://res.slu.se/id/publ/67640