Objective To investigate achaette-scute complex like1/LIM homeobox8 (ASCL1/LIM8) and 19 members of sequence similarity families 4/ microRNA 124-2 (FAM19A4 /miR124-2) in predicting the risk of post-operative recurrence of cervical CIN2/3 grade. Methods A retrospective analysis was performed on 412 patients who were treated for the first time in gynecology department of our hospital from July, 2021 to December, 2022, which were divided into recurrence group and recurrence-free group according to the recurrence situation. The main assessment result was post-operative recurrence, and HPV infection, cytology and colposcopy materials were collected.Results At the end of follow-up, 48 of 412 patients had recurrences, with a recurrence rate of 11.65%. Compared with recurrence-free group, age, age segment, menopausal status, smoking, histopathological diagnosis before treatment, HPV vaccine, ASCL1/LHX8, FAM19A4/miR124-2 were significantly increased in the recurrence group (all P＜0.05). In the recurrence group, methylation levels were obviously higher in recurrence patients with persistent HPV infection compared to with HPV-negative and sporadic HPV infection (all P＜0.05). Among recurrence patients of 48, there were statistically significant differences in ASCL1/LHX8 positive compared with HPV16/18 positivity, cytology positivity and FAM19A4/miR124-2 positivity (all P＜0.05). There were significant differences among cytology, HPV16/28 and FAM19A4/miR124-2, as well as between positive of cytology and HPV16/28 (χ2=17.057, P＜0.001). The median follow-up time was 41 months, there was a statistically significant difference in median recurrence time between recurrence group and recurrence-free group (15vs.41, P＜0.001). Cutoff value (4.75) of risk score was obtained based on Cox method and divided into high and low expression, survival analysis results showed that the recurrence-free rate of patients with high expression was remarkably worse than that of low expression (P＜0.001). Smoking, positive incisal margin, positive of HPV16/18 and cytology at the end of follow-up, positives of ASCL1/LHX8 and FAM19A4/miR124-2, menopause might be the risk factors for recurrence of post-operative in cervical CIN2/3 grade.Conclusion HPV16/18 infection, cytology, ASCL1/LHX8 and FAM19A4/miR124-2 methylation are increased in post-operative patients with cervical CIN2/3 grade, which may affect the risk of recurrence after cervical CIN2/3 grade treatmen