Abstract: Objective To evaluate the application of high-risk human papilloma virus (HPV) E6/E7 mRNA detection method for distribution management of patients with atypical squamous cells undefined significance (ASCUS). Methods A total of 338 patients who were admitted in Lianyungang Maternal and Child Health Hospital from January 2017 to June 2019 for cervical cytological examination and diagnosed with ASCUS were selected.They underwent high-risk HPV DNA genotyping, high-risk HPV E6/E7 mRNA examination and colposcopy biopsy. The correlation between HPV E6/E7mRNA and the pathological results of colposcopy was analyzed. Results The positive rate of high risk HPV E6/E7mRNA gradually increased in the normal group, the LSIL group and the HSIL group (19.27%, 68.72%, and 92.75% respectively), with statistical difference.According to ROC curve, the predictive value of high-risk HPV E6/E7mRNA was better. The positive rate of HPVE6/E7mRNA was 36.3% in 233 cases with positive HPV DNA, which was statistically different from the positive rate of HPV E6/E7mRNA in the HSIL group (95.4%). Conclusions Detection of high-risk HPV E6/E7mRNA is useful for distribution management of ASCUC patients.