[Objective] Screening suitable heat treatment duration and detoxification agents to optimize the efficient virus elimination and regeneration system of tea Chrysanthemum shoot apical meristem, addressing the issue of low rate of obtaining virus-free seedlings. [Methods] Six varieties of tea Chrysanthemum including "WuYuanHuangJu", "JinSiHuangJu", "HangBaiJu", "BeiJingJu", "QiYueXue", and "Ruhe" were used as materials. The axenic seedlings were pre-treated with alternating temperature heat treatment of 40°C/32°C day/night, detoxification agents such as salicylic acid, ribavirin, and abscisic acid, as well as a combination of detoxification agents and heat treatment. The rates of callus induction, regeneration frequency, and average number of regenerated shoots from inoculated shoot apical meristems were compared among different treatment methods, evaluating the detoxification efficacy under various treatments.[Results] (1) There was a significant difference in the regeneration frequency of shoot apical meristems among different varieties of tea Chrysanthemum (P<0.05), with "WuYuanHuangJu" having the highest regeneration frequency of 54.77%, followed by "JinSiHuangJu", "BeiJingJu", "QiYueXue", HangBaiJu and "Ruhe". (2) Under different heat treatment days, the difference in regeneration of shoot apical meristems in tea Chrysanthemum reached a significant level, with "HangBaiJu" and "QiYueXue" showing the best regeneration effect after 40℃/32℃ heat treatment. (3) Salicylic acid, Ribavirin, and Abscisic acid treatments inhibited the regeneration of shoot apical meristems. (4) Salicylic acid and ribavirin treatments combined with heat treatment promoted the regeneration of shoot apical meristems, while high concentration Abscisic acid combined with heat treatment severely inhibited the regeneration of shoot apical meristems. Treatment with 10 μmol/L Salicylic acid combined with heat treatment showed the best regeneration effect on shoot apical meristems. (5) Obtaining completely virus-free seedlings through heat treatment, the combined method of detoxification agents and heat treatment. [Conclusion] The combination of 10 μmol/L salicylic acid concentration with alternating temperature of 40°C/32°C day/night pre-treatment for 40 days significantly enhances the regeneration frequency of shoot apical meristems and results in obtaining virus-free seedlings.