P16/Ki67 Dual Staining And High Risk HPV E6/E7 MRNA Detection In Cervical Cancer Screening

Background and PurposeCervical cancer is one of the gynecological tumors which is harmful for women’s health and life safety.With the improvement of people’s educational level,the progress of medical level and the positive promotion of cervical cancer screening,the detection of cervical lesions in China has been significantly improved.Cytology is a traditional method in cervical cancer screening,with high specificity and much higher accuracy than Pap smear technology.However,its sensitivity is not high,because it has to make subjective judgment according to the shape of cells,as well as the interference of the factors such as the production and staining,and the difference of the professional level of different cytologists,which may lead to its insufficient diagnostic efficiency。High risk human papillomavirus(HPV)detection is a revolutionary change from morphological detection to molecular biological detection with high sensitivity.However,HPV detection can not distinguish persistent infection or not,so the false positive rate will increase and the misdiagnosis rate will increase。Therefore,many people are exploring some new detection methods to improve thescreening efficiency of cervical cancer.HPV E6/E7 mRNA and p16/Ki67 dual staining are two new biomarkers.P16/Ki67 dual staining method is a combined biomarker that can indicate HPV transforming infection.When p16 and Ki67 are found in the same cervical epithelial cell,it means that the cell is in an abnormal proliferation state,and the original cycle has been destroyed.This may indicate the occurrence of high-grade cervical squamous intraepithelial lesions.HPV E6/E7 mRNA can identify whether HPV infection is a primary infection.When E6,E7 protein is in low expression,it is proved that HPV is not continuous infection,and it will not produce cervical high lesions.However,when HPV infection continues,there are a large number of mRNA transcription and translation.Then the cycle of cells changes into malignant tumor cells,and cervical lesions occur.The purpose of this study was to explore the efficacy of p16/Ki67 dual staining and HPV E6/E7 mRNA detection methods in cervical cancer screening,and to provide a new idea for cervical cancer screening.MethodsThe samples of patients who received cervical cancer screening in the First Affiliated Hospital of Zhengzhou University from October 2018 to October 2020 were collected.There are relevant inclusion and exclusion criteria.A total of 745 patients were enrolled,with an average age of 42.46 ±11.05 years old(19-70 years old).TCT,high-risk HPV,p16/Ki67 dual staining,E6/E7 mRNA detection and pathological results of colposcopy biopsy were detected in each specimen.SPSS 25.0 and medcalc software were used to calculate the sensitivity,specificity,positive predictive value,negative predictive value,and area under ROC curve(AUC).Measurement data were analyzed by t-test and Kruskal Wallis analysis,P<0.05 was statistically significant.A total of 745 patients were collected,including 218 patients with chronic cervical inflammation,138 patients with LSIL,298 patients with HSIL,2 patients with adenocarcinoma in situ and 89 patients with cervical cancer.Results1.Among 745 patients,the sensitivity of HPV detection(96.92%)was significantly higher than that of E6/E7(73.01%)(P<0.01);but its specificity(8.71%)was far lower than that of E6/E7(42.98%)(P<0.01).The sensitivity of TCT was significantly higher than that of E6/E7(73.01%)(P<0.05),but the specificity(39.60%,42.98%)was not significantly different(P>0.05);The sensitivity of p16/ki67 test was significantly lower than that of HPV(96.92%)(P<0.01),but its specificity(84.27%)was significantly higher than that of HPV(8.71%)(P<0.01).The sensitivity of p16/ki67(81.49%)was not significantly different from that of TCT(80.21%)(P>0.05),but its specificity(84.27%)was significantly higher than that of TCT(39.60%)(P<0.01).The sensitivity(81.49%)and specificity(84.27%)of p16/ki67 were significantly higher than that of E6/E7(73.01%,42.98%)(P<0.05).The AUC of E6/E7 test(0.580)showed no significant difference compared with that of HPV test(0.528)and TCT test(0.599)(P>0.05).The AUC of double staining test(0.829)was significantly higher than that of HPV test(0.528),TCT test(0.599)and E6/E7 test(0.580)(P<0.05).2.when triage other high-risk HPV positive patients(300 cases),the sensitivity of p16/ki67 test(78.07%)was not significantly different from that of TCT(86.84%),and the specificity(82.26%)was significantly higher than that of TCT(29.57%)(P<0.01).The sensitivity of E6/E7 was significantly lower than that of TCT(86.84%)(P<0.05),and there was no significant difference between specificity(30.11%)and TCT(29.57%)(P>0.05).The sensitivity of p16/ki67(78.07%)was not significantly different from that of E6/E7(73.68%)(P>0.05);the specificity(82.26%)was significantly higher than that of E6/E7(30.11%)(P<0.05).There was no significant difference between the AUC of E6/E7 test(0.519)and TCT test(0.582)(P>0.05).The AUC of double staining test(0.802)was significantly higher than that of TCT test(0.582)and E6/E7 test(0.519)(P<0.05).3.In 196 patients with ASCUS,the sensitivity of dual staining detection(77.32%)was not significantly different from that of E6/E7 detection(67.01%)(P>0.05);the specificity(84.85%)was significantly higher than that of E6/E7 detection(39.39%)(P<0.05).The sensitivity of p16/Ki67 detection(77.32%)was significantly lower than that of high-risk HPV detection(98.97%)(P<0.01);the specificity(84.85%)was significantly higher than that of HPV detection(11.11%)(P<0.05).The sensitivity of E6/E7 detection(67.01%)was significantly lower than that of HPV(98.97%)(P<0.01);the specificity(39.39%)was significantly higher than that of HPV(11.11%)(P<0.01).The AUC of E6/E7 test(0.532)was not significantly different from that of HPV test(0.550)(P>0.05).The AUC of dual stain test(0.811)was significantly higher than that of HPV test(0.550)and E6/E7 test(0.532)(P<0.05).4.There was no significant difference between the sensitivity of p16/Ki67(78.72%)and E6/E7 detection(80.85%)in 127 patients with LSIL(P>0.05);the specificity(85.00%)was significantly higher than that of E6/E7 detection(33,75%)(P<0.05).The sensitivity of p16/Ki67 detection(78.72%)was significantly lower than that of HPV detection(95.74%)(P<0.05);the specificity(85.00%)was significantly higher than that of HPV detection(16.25%)(P<0.05).The sensitivity of E6/E7 detection(80.85%)was significantly lower than that of HPV(95.74%)(P<0.05);the specificity(33.75%)was significantly higher than that of HPV(16.25%)(P<0.05).There was no significant difference between the AUC of E6/E7 test(0.573)and HPV test(0.560)(P>0.05).The AUC of dual stain test(0.819)was significantly higher than that of HPV test(0.560)and E6/E7 test(0.573)(P<0.05).Conclusions1.As two new biomarkers,p16/Ki67 test and E6/E7 test showed good efficacy in cervical cancer screening,among which p16/Ki67 test performed better than the other three screening methods;the screening efficacy of E6/E7 test was similar to that of HPV and TCT test.2.In triage other HPV positive patients,p16/Ki67 detection was significantly better than TCT detection and E6/E7 detection.The sensitivity of p16/Ki67 and E6/E7 detection was slightly lower than that of TCT detection,but the specificity of p16/Ki67 was much higher than that of TCT and E6/E7 detection.The overall screening efficiency of p16/Ki67 was still the highest.3.In ASCUS patients,the sensitivity of p16/Ki67 was similar to that of E6/E7 detection,which was slightly lower than that of HPV detection,but the specificity of p16/Ki67 was much higher than that of E6/E7 detection and HPV detection;in LSIL patients,the results were the same.