Positive human leukocyte antigen B27 (HLA-B27) gene assay is associated with ankylosing spondylitis, but does not confirm the diagnosis of ankylosing spondylitis. HLA-B27 belongs to the major histocompatibility complex genes, which are mainly expressed on human lymphocytes and other nucleated cells. Since a large amount of data shows that there is a link between HLA-B27 positivity and ankylosing spondylitis, patients with positive HLA-B27 need to further improve the relevant auxiliary examinations such as nuclear magnetic resonance imaging (NMRI), CT, autoimmune antibodies, and other relevant examinations to clarify the presence of ankylosing spondylitis. However, a positive HLA-B27 test does not necessarily indicate ankylosing spondylitis, but can only provide a clue for the final diagnosis, and a negative test does not completely exclude ankylosing spondylitis. In addition, there is a possibility that the offspring of a patient with a positive HLA-B27 test may carry the same disease-causing gene. It is recommended that patients with positive HLA-B27 gene test should go to the rheumatology and immunology department of regular hospitals to further improve the relevant auxiliary examinations to confirm the diagnosis.