Evaluation of A Novel Point‐of‐Care Real‐Time PCR Platform for Rapid Screening of HLA ‐ B27 Using Whole Blood

HLA-B27 positivity is a key determinant for early referral of axial spondyloarthropathies, yet timely testing remains constrained by reliance on centralised molecular laboratories requiring substantial infrastructure and specialised personnel. We developed and validated a fully automated, cartridge-based real-time PCR point-of-care platform, the MoiM HLA-B27 Detection Kit run on the MoiM Dx100 analyser, which integrates sample loading, magnetic bead-based DNA extraction, and qPCR into a single disposable cartridge for sample-to-answer testing directly from whole blood. Analytical performance was characterised for extraction efficiency, limit of detection, and specificity using the UCLA Immunogenetics Center DNA Reference Panel, and clinical performance was evaluated in 217 whole blood specimens submitted for routine HLA-B27 genotyping, compared against a reference PCR sequence-specific primer (PCR-SSP) method with discordant cases arbitrated by Luminex PCR-SSOP. The platform produced DNA of purity comparable to manual extraction (mean A260/A280 1.9 ± 0.04), with 100% analytical specificity and no cross-reactivity against B7-related cross-reactive group alleles. Optimising the cycle threshold cut-off eliminated non-specific late-cycle amplification responsible for four initial false positives. In the final clinical cohort (67 positive, 150 negative), the assay achieved 100% concordance with PCR-SSP (sensitivity 100% [95% CI 94.6-100%]; specificity 100% [95% CI 97.6%-100%]; Cohen's κ = 1.00), with shorter hands-on time, faster turnaround, and lower per-test cost. This integrated platform delivers laboratory-grade HLA-B27 results in a rapid, accessible format that may shorten the diagnostic pathway for ankylosing spondylitis and related spondyloarthropathies.