Objective: To investigate the application value of human papillomavirus (HPV) E6/E7 mRNA combined with thinprep cytologic test (TCT) in cervical cancer screening. Methods: A total of 975 patients with cervical cancer who underwent cervical cancer screening and obtained pathological results in Zhangjiakou Maternal and Child Health Hospital from January to July 2021 were selected as the study subjects. All subjects underwent HPVE6/E7 mRNA and TCT. According to the screening results, colposcopy cytology was performed in the positive cases of HPVE6/E7 mRNA and TCT. Taking histopathological results as the gold standard, the value of HPVE6/E7 mRNA, TCT, and their combination in the diagnosis of cervical lesions was compared. Results: Among the 975 patients, there were 352 chronic cervicitis, 315 low-grade squamous intraepithelial lesion (LSIL), 263 high-grade squamous intraepithelial lesion (HSIL) and 45 cervical cancer. The positive rate of HPVE6/E7 mRNA increased with the grade of cervical lesions (P<0.05). The efficacy of HPVE6/E7 mRNA in the diagnosis of HSIL and above cervical lesions was higher than that of TCT (P<0.05). The efficacy of HPVE6/E7 mRNA combined with TCT in the diagnosis of HSIL and above cervical lesions was higher than that of HPVE6/E7 mRNA and TCT alone (P<0.05). Conclusion: The combined detection of HPVE6/E7 mRNA and TCT can improve the diagnostic value of HSIL and above cervical lesions, and plays an important role in cervical cancer screening.