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, 95 (3), 189-195

Real-time PCR Based HLA-B*27 Screening Directly in Whole Blood

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Real-time PCR Based HLA-B*27 Screening Directly in Whole Blood

Kathrin Geiger et al. HLA.

Abstract

The linkage between the occurrence of human leucocyte antigen B*27 (HLA-B*27) and ankylosing spondylitis or other related spondyloarthritides is well documented. PCR based methods are widely used for HLA-B*27 screening. To refine HLA-B*27 testing we aimed at establishing a real-time PCR protocol to detect the HLA-B*27 allele directly in blood samples, without DNA extraction. HLA-B*27 analysis was performed by two real-time PCRs using TaqMan primer-probe assays for B*27 specific amplification of exon 2 or exon 3 of the HLA-B gene together with a mutant of Taq polymerase for direct blood PCR. Conditions for direct blood PCR were optimized and the reliability of the direct blood PCR protocol was evaluated by re-genotyping over 200 blood samples from patients who previously underwent routine DNA-based HLA-B*27 testing. Heating blood samples at 95°C for 10 minutes significantly improved PCR performance. Results from real-time PCR based HLA-B*27 testing directly in blood of over 200 patients were in 100% concordance with results obtained by routine DNA-based HLA-B*27 genotyping. In summary, we present a reliable real-time PCR protocol for HLA-B*27 screening directly in whole blood supporting fast clarification of the presence of ankylosing spondylitis or other spondyloarthritides in suspected cases.

Keywords: HLA-B*27; ankylosing spondylitis; direct blood PCR; real-time PCR.

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References

REFERENCES

    1. Chen B, Li J, He C, et al. Role of HLA-B27 in the pathogenesis of ankylosing spondylitis. Mol Med Rep. 2017;15(4):1943-1951. https://doi.org/10.3892/mmr.2017.6248.
    1. Bodis G, Toth V, Schwarting A. Role of human leukocyte antigens (HLA) in autoimmune diseases. Rheumatol Ther. 2018;5(1):5-20. https://doi.org/10.1007/s40744-018-0100-z.
    1. McMichael A, Bowness P. HLA-B27: natural function and pathogenic role in spondyloarthritis. Arthritis Res. 2002;4(Suppl 3):S153. https://doi.org/10.1186/ar571.
    1. Reveille JD. Major histocompatibility genes and ankylosing spondylitis. Best Pract Res Clin Rheumatol. 2006;20(3):601-609. https://doi.org/10.1016/j.berh.2006.03.004.
    1. Steffens-Nakken HM, Zwart G, van den Bergh FA. Validation of allele-specific polymerase chain reaction for DNA typing of HLA-B27. Clin Chem 1995;41(5):687-692. http://www.ncbi.nlm.nih.gov/pubmed/7729046.

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