The invention relates to a method for rapidly detecting a human leucocyte antigen B27 (HLA-B27) and an in-vitro diagnosis kit, which belong to the technical field of medical biology. In the kit, a pair of HLA-B27 specific primers and a specific fluorescent probe are adopted for rapidly detecting the HLA-B27. In the kit, the pair of HLA-B27 specific primer and the specific fluorescent probe can beused for rapidly and accurately detecting HLA-B27 genes in samples such as human peripheral blood and the like, and the kit has the advantages of high specificity, high sensitivity, time saving, labor saving, low cost, high flux and the like, and can be applied to auxiliary diagnosis of multiple spondyloarthropathies which severely endanger human health such as clinical ankylosing spondylitis, Reiter syndrome, psoriatic arthritis, ulcerative colitis accompanying arthropathy, acute anterior tract uveal, and the like.