A homogeneous HLA-B*27 genotyping assay using dried reagent mixtures

Area of Science:

• Immunogenetics
• Molecular Diagnostics
• Rheumatology

Background:

• The human leukocyte antigen B*27 (HLA-B*27) allele is strongly associated with ankylosing spondylitis.
• Accurate and accessible diagnostic methods for HLA-B*27 are crucial for patient management.

Purpose of the Study:

• To develop and validate a novel, simplified assay for HLA-B*27 typing.
• To assess the performance and stability of an assay using dried reagents and homogeneous time-resolved fluorescence PCR.

Main Methods:

• Development of a homogeneous time-resolved fluorescence based PCR assay for HLA-B*27 typing.
• Utilized whole blood samples dried on collection cards with pre-dried reagent wells.
• Assay validation involved comparison with a DELFIA-based method on 229 patient samples.

Main Results:

• The developed assay demonstrated 100% concordance with the established DELFIA-based method.
• Dried reagents remained stable for at least eight weeks at room temperature without performance degradation.
• The assay requires only the addition of the sample to initiate the homogeneous amplification.

Conclusions:

• A novel, simplified, and stable assay for HLA-B*27 typing has been successfully developed and validated.
• This assay offers a promising tool for the diagnostic process of ankylosing spondylitis.
• The use of dried reagents enhances assay convenience and stability for clinical applications.