Objective：This study aims to explore the predictive value of p16/Ki-67 in the progression of high-risk HPV-positive patients with negative cytology. Methods：One hundred and sixty-eight high-risk HPV-positive women with negative cytology were enrolled from May 2017 to June 2017 in the Department of Gynecology，the First Affiliated Hospital of Nanjing Medical University. Follow up were conducted for a period of 2 years to analyze the relationship between the progression and p16/Ki-67 dual staining. Results：①Outcome：88 cases（52.4%）had regressed，54 cases（32.1%）persisted，26 cases（15.5%）progressed；②Univariate analysis showed that age of patient，HC2 HPV value and high-risk HPV genotyping were not risk factors of the progression（P＞0.05），positive p16/Ki-67 dual staining showed higher rates of progression（P＜0.001）. Multivariable logistic analysis showed that positive p16/Ki-67 dual staining was related to the progression；③55 cases had positive p16/Ki-67 dual staining，19 cases（34.5%）of them progressed；113 cases had negative p16/Ki - 67 dual staining，7 cases（6.2%）of them progressed. There were significant difference in progression between patients with positive p16/Ki-67 dual staining and patients with negative p16/Ki-67 dual staining（P＜0.05）. The sensitivity of p16/Ki-67 in prognosis was 73.1%，the specificity was 74.7%，the positive predictive value was 34.5%，the negative predictive value was 93.8%；④High-risk HPV natural changes：152 patients finished the 2-year follow up. The negative conversion ratio at 6，12，18，24 months for women with negative results of p16/Ki-67 dual staining were higher than those with positive results（P＜0.05）. Conclusion：Positive p16/Ki-67 dual staining result was a risk factor for progression and negative high-risk HPV conversion. P16/Ki-67 dual staining had predictive value for progression and showed good risk stratification for TCT-negative and high-risk HPV-positive women.