The rapid detection of human HLA-B*27 gene based on BASIC isothermal detection

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Immunogenetics

Background:

• Ankylosing spondylitis (AS) is a chronic inflammatory arthritis linked to the human leukocyte antigen B27 (HLA-B27).
• Current HLA-B27 detection methods like qPCR and flow cytometry are time-consuming and labor-intensive.
• A need exists for a rapid and efficient diagnostic tool for HLA-B27 detection.

Purpose of the Study:

• To develop a rapid diagnostic platform for HLA-B27 gene detection.
• To combine isothermal amplification (BASIS) with CRISPR/Cas12a for enhanced specificity and speed.
• To evaluate the platform's performance against existing methods for ankylosing spondylitis screening.

Main Methods:

• Developed the BASIC platform integrating BASIS isothermal amplification and CRISPR/Cas12a.
• Utilized universal primers targeting conserved regions for broad HLA-B*27 genotype amplification.
• Employed CRISPR/Cas12a with specific guide RNA for precise detection of HLA-B*27 amplicons.

Main Results:

• The BASIC platform achieved detection in 1 hour with analytical sensitivity as low as 100 aM.
• Demonstrated resistance to interference from homologous genes and common biological substances.
• Clinical sample detection showed complete concordance with quantitative real-time PCR (qPCR).

Conclusions:

• The BASIC platform is a sensitive, specific, simple, and rapid tool for HLA-B27 gene detection.
• It shows significant promise for early screening and diagnosis of ankylosing spondylitis.
• Offers a viable alternative to conventional methods, improving diagnostic efficiency.