The Study Of The Effect Of DNA Methylation Assay In Triaging High-risk HPV-positive Women

Objective:Cervical cancer is a common malignant tumor that seriously endangers women’s health.The persistent infection of high-risk HPV is an important reason for the occurrence and development of cervical cancer.HPV detection has gradually replaced cytological detection as the primary screening method for cervical cancer due to its higher detection sensitivity.The relatively low specificity of HPV testing leads to unnecessary diagnosis and treatment for many women,which increases medical costs.Therefore,it is urgent to find effective triage methods.Currently,the common triage methods for high-risk HPV-positive women include cytology test and HPV16/18 genotyping test.Both have disadvantages in clinical application,so it is necessary to find other efficient triage methods.The aim of this study is to evaluate the efficacy of multiple methods,including ASTN1、DLX1、ITGA4、RXFP3、SOX17、ZNF671 host gene methylation assay,alone and in combination in triage women with high-risk HPV infection.Methods:Women who had received abnormal cervical cancer screening results and obtained pathological results of the cervical biopsy under colposcopy in the gynecological outpatient department of Shengjing Hospital of China Medical University from September 2021 to September 2022 were selected for the study.A total of 243 high-risk HPV-positive women were included in this study,and a DNA methylation test was performed on the discarded TCT remaining samples of the enrolled women.The triage performance of the DNA methylation test,TCT test,HPV16/18 genotyping test,visual inspection with acetic acid,visual inspection with Lugol’s iodine alone,and combined detection on high-risk HPV-positive women was compared.During the study,P<0.05 was used as the test level.Results:1.The top five high-risk HPV types that were more frequently infected in this study were 16(41.6%),58(15.2%),52(14.8%),18(11.1%),and 51(9.1%).2.The differences in host gene methylation detection,VIA detection,VILI detection,and HPV16/18 genotyping detection between the CIN1-group and the CIN2+group were statistically significant(P<0.05).There was no significant difference in the positive rate of TCT detection between the two groups(X~2=0.214,P=0.643).3.The differences in host gene methylation detection,TCT detection,VIA detection,and HPV16/18 genotyping detection between the CIN2-group and the CIN3+group were statistically significant(P<0.05).There was no significant difference in the positive rate of VILI detection between the two groups(X~2=2.512,P=0.113).4.With the increase of cervical lesions,the positive rates of host gene methylation,VIA,and VILI all showed an increasing trend.The positive rate of VILI in different grades of cervical lesions was higher than that of other detection methods.With the increase in cervical lesion grade,the positive rate of the TCT test and HPV16/18 genotyping test did not increase.5.Among all the triage methods used alone to detect CIN2+/CIN3+women,VILI had the highest sensitivity and negative predictive value,and VIA had the highest specificity,positive predictive value,and accuracy.The specificity,positive predictive value,negative predictive value,and accuracy of host gene methylation in detecting CIN2+/CIN3+women were higher than those of TCT and HPV16/18 genotyping.6.The AUC of host gene methylation for detecting CIN2+women was 0.73,and the AUC of host gene methylation for detecting CIN3+women was 0.85,both of which were higher than those of TCT for detecting CIN2+women and CIN3+women.7.When the two triage methods were used as the combined detection method,the sensitivity of the combined detection method was higher than that of the two detection methods alone,and the specificity of the combined detection method was lower than that of the two detection methods alone.When the two detection methods are used to detect CIN2+and CIN3+patients,VIA combined with host gene methylation detection has the highest detection specificity while ensuring the detection sensitivity.8.With the aggravation of cervical lesions,except for ITGA4 methylation detection indicators,the positive rates of the other five single host gene methylation detection indicators showed an increasing trend.DLX1 methylation had the highest positive rate among the six methylation markers,while ITGA4 methylation had the lowest positive rate.9.When single host gene methylation was detected in CIN2+and CIN3+women,the DLX1 methylation assay had the highest detection sensitivity,66.2%,and 81.3%,respectively.ITGA4 methylation assay had the lowest sensitivity but the highest specificity and positive predictive value:specificity:97.1%and 95.3%, respectively;positive predictive value:73.7%and 47.4%,respectively.Conclusion:1.Host gene methylation detection,VIA detection,and HPV16/18genotyping detection can be used to identify the difference between the low-grade cervical intraepithelial lesion and the high-grade cervical intraepithelial lesion.2.Host gene methylation,VIA,and VILI detections are predictive of cervical lesion grade.3.The specificity,positive predictive value,negative predictive value,and accuracy of DNA methylation testing in the diagnosis of CIN2+/CIN3+were better than those of TCT and HPV16/18 genotyping testing in triage high-risk HPV-positive women.4.When any two methods were randomly combined to triage high-risk HPV-positive women,the diagnostic sensitivity increased while the specificity decreased.5.Host gene methylation detection has efficacy in triaging high-risk HPV-positive women.