Multiplexed fluorometric immunoassay testing methodology and troubleshooting

Area of Science:

• Veterinary diagnostics
• Biomedical research quality assurance
• Infectious disease detection

Background:

• Laboratory animal quality is crucial for reliable biomedical research.
• Serological testing is the standard for monitoring animal health.
• Traditional indirect enzyme-linked immunosorbent assay (ELISA) is a singleplex method for serosurveillance.

Purpose of the Study:

• To develop advanced diagnostic testing strategies for adventitious infectious agents in animals.
• To introduce a multiplexed assay for enhanced serosurveillance.
• To improve the efficiency and accuracy of animal model quality control.

Main Methods:

• Development of a multiplexed fluorescence immunoassay (MFIA) using microspheres.
• Simultaneous detection of up to 100 assays in a single well.
• Incorporation of internal control beads (tissue control, IgG anti-test serum species immunoglobulin, IgG control) for validation.

Main Results:

• The MFIA allows for up to 100 simultaneous assays, significantly increasing information yield per well.
• Reduced requirements for serum, reagents, and disposables compared to singleplex assays.
• Internal controls ensure sample suitability and system functionality, validating results.

Conclusions:

• The MFIA represents a significant advancement in serosurveillance for laboratory animals.
• This multiplexed approach enhances diagnostic efficiency, reduces costs, and improves the reliability of animal model data.
• The validated methodology ensures higher quality animal models for biomedical research.