Objective To investigate the clinical features,electrophysiological results,treatment regimen,and prognosis of anti-GAD65 antibody-associated stiff-person syndrome. Methods A retrospective analysis was performed for four patients with anti-GAD65 antibody-associated stiff-person syndrome who attended Department of Neurology,The First Affiliated Hospital of Air Force Medical University,from December 2019 to March 2023. Clinical phenotype,electrophysiological characteristics,treatment methods and prognosis were analyzed. Results All four patients with anti-GAD65 antibody-associated stiff-person syndrome were female,with an age of 49-74 years,and delayed diagnosis(1-7 years) was observed in all four patients. Among the four patients,one had epilepsy,one had cerebellar ataxia,one had type 1 diabetes,and three had Hashimoto's thyroiditis. In the acute stage of the disease,one patient received high-dose hormone shock therapy,one received human immunoglobulin therapy,and two received low-dose hormone. As for long-term immunotherapy,three patients were given mycophenolate mofetil,and one patient was given regular infusion of human immunoglobulin. All four patients showed response to clonazepam and immunoregulatory therapy. Conclusion Anti-GAD65 antibody-associated stiff-person syndrome is often observed in middle-aged and elderly women,with delayed diagnosis in most cases. Patients may also have other clinical phenotypes and autoimmune diseases,and typical discharge of motor unit potential often exits in axial muscles. Clonazepam and immunotherapy are effective treatment methods for this disease.