〔Abstract〕 Objective To analyze the clinical research value of hematological indexes such as human leukocyte antigen B27 (HLA-B27), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and disease activity in the auxiliary diagnosis of ankylosing spondylitis (AS). Methods 129 patients with AS (AS group) treated in the Second People's Hospital of Guangdong Province from December 2015 to January 2019 were randomly selected for retrospective analysis. All patients were tested for HLA-B27, ESR, CRP and disease activity evaluation; 52 normal subjects (healthy group) who underwent physical examination in our hospital in the same period were randomly selected as the control. Results The positive rates of HLA-B27, ESR and CRP in AS group were 85.27%, 69.77% and 56.59% respectively, which were significantly higher than 3.85%, 9.62% and 0.00% in healthy group (P < 0.05). According to the ankylosing spondylitis disease activity score (ASDAS), AS patients were divided into low disease activity group (66 cases) and high disease activity group (63 cases). The overall evaluation of patients and doctors, evaluation of whether to receive intensive treatment, and Bath ankylosing spondylitis disease activity index (BASDAI) scores in the high disease activity group were higher than those in the low disease activity group, The differences were statistically significant (P < 0.05). Conclusion HLA-B27 index can be used for clinical auxiliary diagnosis of AS. Its combination with ESR and CRP has certain guiding significance for analyzing the disease activity, disease development and prognosis of AS patients.