【Abstract】Objective The objective of this essay was to analyze the diagnostic value of cytokeratin 7 (CK7), p16 and serum squamous cell carcinoma antigen (SCC-Ag) in the diagnosis of squamous intraepithelial lesion (SIL). Methods 264 patients in Department of Gynecology, Handan Central Hospital from August 2018 to December 2019 were selected as the study subjects. According to the pathological diagnosis results, 264 patients were divided into control group, low-level cervical squamous intraepithelial lesions (LSIL) group and high-level cervical squamous intraepithelial lesions (HSIL) group. 77 cases of cervical biopsy inflammation were treated as control group, 92 cases in LSIL group and 95 in HSIL group. The expression of CK7 and p16 in each group was detected by immunohistochemistry SP method. The expression of SCC Ag in serum of each group was detected by chemiluminescence immunoassay. The expression of CK7, SCC Ag and p16 in each group was analyzed by the association coefficient. The sensitivity, specificity, positive predictive value, negative of single test, two pair test and combined test were observed. The accuracy of the predicted value and the Jordan index were compared with the diagnostic effectiveness of the three. Results In control group, LSIL group and HSIL group, the positive expression rates of CK7, p16 and SCC Ag increase gradually, which are 10.39%, 31.52%, 67.37%, 5.19%, 48.91%, 95.79%, 6.49%, 15.22%, 26.32%, respectively. The differences between the HSIL group and other groups were statistically significant (χ2=53.241, P＜0.05; χ2=108.933, P＜0.05; χ2=9.948, P＜0.05). CK7 was positively correlated with SCC Ag (r=0.204, P=0.001), CK7 was positively correlated with p16 (r=0.437, P=0.000), and SCC Ag was positively correlated with p16 (r=0.350, P=0.000). The sensitivity and negative predictive value of the three methods were the highest (98.95%, 99.00%). The three had obvious positive synergy effect, and the sensitivity and negative predictive value of the three combined detection were the highest (98.95%, 99.00%). Conclusion The combined detection of CK7, SCCAg and P16 protein has better diagnostic value than single or pairwise paired detection. The combined detection of CK7, SCC Ag and p16 may be one of the value indicators of SIL diagnosis.