Objective：To investigate the clinical value and significance of high risk human papilloma virus（HPV）E6/E7 mRNA test in patients with HPV infection. Methods：Between January 2017 and June 2017，in Center of Diagnosis and Treatment for Cervical Disease，200 patients with HPV infection in cervical screening received high-risk HPV E6/E7 mRNA test and pathological biopsy under colposcopy. We analyzed the E6/E7 mRNA detection rate of positive cases，and the E6/E7mRNA expression in different pathological diagnosis. Results：The E6/E7 mRNA detection rate was 60% in the 200 patients. The highest E6/E7 mRNA positive expression in the high-grade squamous intraepithelial lesion group（HSIL）had significant differences with the low-grade squamous intraepithelial lesion group（LSIL）and the nomal group. The mRNA sensitivity was 97.9% and negative predictive value was 98.8%（P < 0.05）. The sensitivity，specificity，positive predictive value and negative predictive value were lower in the LSIL group. Conclusion：We could shunt the patients with HPV infection，avoid excessive treatment and reduce the patient’s psychological burden byHR-HPV E6/E7 mRNA test. High-risk HPV E6/E7 mRNA test has high clinical value in diagnosis of HSIL during cervical screening.