Objective：To explore the laboratory verification methods and clinical characteristics of cytological HPV-negative cervical cancer. Methods：From September 2015 to October 2018，33 cervical cancer patients with HPV-negative and pathologically diag－nosed by cervical exfoliated cell Hybribio 21 typing（K-21） and 60 HPV-positive cervical cancer patients in the same period were collected. Laboratory validation was carried out with pathological paraffin specimens，molecular pathology（hybribio membrane hybridization + PCR） kits（K-14，K-18 and K-37）. To analyze the clinical characteristics of HPV-negative and positive cervical cancer. Results：33 cases of cervical cancer with negative cytology were detected by tissue paraffin section and molecular pathology. The results of K-14 and K-18 kits were 13 negative and 9 of K-37 kits were negative. There were statistical differences in the first clinical manifestations between 9 patients with negative cervical cancer and 84 patients with positive cervical cancer in molecular pathology，the proportion of contact bleeding and irregular vaginal bleeding in negative group was smaller than that in positive group（3/9∶41/84，1/9∶33/84； ?字2=12.889，P=0.006）. There were statistical differences in FIGO stages at the time of treatment，the proportion of Ib2，IIa2，Ⅲ and Ⅳ stages in negative group was higher than that in positive group（3/9∶13/84，1/9∶7/84，2/9∶0/84； ?字2=12.786，P=0.021）. There were statistical differences in pathological types，the proportion of adenocarcinoma in negative group was higher than that in positive group（3/9∶5/84； ?字2=5.752，P=0.041）. There were statistical differences in tumor infiltration depth among surgical patients，the propor－tion of early infiltration，infiltration to cervical external orifice and infiltration to uterine body in negative group was higher than that in positive group（2/7∶6/64，2/7∶14/64，3/7∶11/64； ?字2=8.332，P=0.035）. Conclusion：Theoretically，HPV-negative cervical cancer exists. Clinically，HPV negative cervical cancer has the characteristics of atypical initial clinical manifestations，late FIGO stage and large proportion of adenocarcinoma.