Sammanfattning

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.

Nyckelord

Acoustic trapping; Immuno-based assay; Multiplex; Luminex

Publicerad i

Analytica Chimica Acta
2015, Volym: 853, sidor: 682-688

SLU författare

• Xia, Hongyan

  • Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
    
  • World Organization for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine

• Belak, Sandor

  • Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
    
  • Sveriges Veterinärmedicinska Anstalt (SVA)
  • World Organization for Animal Health (OIE) Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine

UKÄ forskningsämne

Medicinsk biovetenskap


Publikationens identifierare

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

Permanent länk till denna sida (URI)

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