The Study Of The Application Of Human Papilloma Virus E6/E7 MRNA Detection In Cervical Cancer Screening

Background and objective:Cervical cancer is one of the common gynecological malignancies,from the progressive development of it?s precancerous lesions.It is a long and gradual process from CIN to cervical cancer.So,efficient screening in the prevention and treatment of cervical cancer is particularly important.Numerous studies show that HPV infection is the most common cause of cervical cancer and its precancerous lesions.Currently,TCT combined with HPV detection is a major clinical screening tool.TC T inspection process due to operation,reading and other subjective factors prone to false negative or false positive.HPV DNA detection is currently widely used HPV detection technology.But most of the HPV infection in clinical lack of continuity and stability.Most new infections can fade within 2 years.This makes the specificity of HPV DNA detection relatively low.Means that HPV DNA detection methods may detect a large number of clinically meaningless infections.Thereby increasing the patient's economic and psychological burden.This study was designed to investigate the value of HPV E6/E7 mRNA detection in HPV-positive patients,and to explore the application value of HPV E6/E7 mRNA in cervical cancer screening.Materials and methods:1.Clinical information: Selected in June 2015-October 2016 in Jilin University First Hospital Cancer Gynecological outpatient department and inpatient department of 200 cases of TCT and HPV DNA test confirmed HPV positive patients.Patients aged 23 to 72 years old,average age(40.50 ± 10.50)years old.No history of hysterectomy and cervical surgery,no history of pelvic radiation therapy,3 days of asexual life,vaginal washing and vaginal drug operation.TCT cytology classification:No intraepithelial lesion or malignant cells(NILM)110 cases,Atypical squamous cells(ASC)29 cases,Low-grade squamous intraepithelial lesions(LSIL)23 cases,High-grade squamous intraepithelial lesions(HSIL)27 cases,Squamous cell carcinoma or Atypical glandular cells(SCC/AGC)11 cases.Among the 200 patients,100 cases were examined by cervical biopsy and pathology.Biopsy Pathology Category: No intraepithelial lesion or malignant cells(NILM)20 cases,CIN grade I28 cases,CIN grade? 39 cases,CIN grade ? 8 cases,Cervical cancer 5 cases.All patients signed informed consent.2.Method2.1 Specimen collection and preservation :With a special cervical brush into the cervix outside the mouth,turn clockwise 3 to 5 turns.Collect cervical surface and cervical exfoliated cells.The cervix was then shaken 10 times in a bottle containing Preserv Cyt TM stock solution.The cells after the elution of the lid,paste the label,minus20? to keep the preparation.2.2 Specimens were:Load the specimens into a refrigerator,then send to the Henan Cotech(Xinxiang)Biotechnology Co.,Ltd.2.3 HPV E6/E7 mRNA detection: Researchers of the company strictly follow the HPV E6/E7 mRNA detection instructions for testing.2.4 Settle results:The results of HPV E6/E7 mRNA of the 200 specimens and the results of TCT tests and cervical biopsy are all recorded in Excel to do statistical analysis.3.Statistical analysis :This study is a cross-sectional survey design.The data were analyzed using SPSS 19.0 software.The positive trend of HPV E6/E7 mRNA positive rate in different grades of cervical lesions was analyzed by univariate linear regression analysis.HPV E6/E7 mRNA expression levels are expressed in quartiles P75.Comparison of HPV E6/E7 mRNA expression levels in cervical lesions at different levels Spearman correlation analysis was used to show the changes in HPV E6/E7 mRNA copy number with cervical lesions.P <0.05 for the difference is statisticallysignificant.Results:1.There were significant differences in HPV E6/E7 mRNA positive rate between different TCT grade cervical lesions.And HPV E6/E7 mRNA positive rate and TCT grade was positively correlated(F value= 9.474,P=0.037<0.05,Positive ratio of the prediction coefficient is 0.955).There was also a positive correlation between HPV E6/E7 mRNA copy number and TCT grade(P=0.002778<0.05,spearman rank correlation coefficient is 0.943).2.There were significant differences in HPV E6/E7 mRNA positive rate between different pathological grades of cervical lesions.And the positive rate of HPV E6/E7 mRNA was positively correlated with pathological grade(F value=33.563,P=0.010<0.05,Positive ratio of the prediction coefficient is 0.952).HPV E6/E7 mRNA copy number and pathological grading also showed a positive correlation(P=0.01667<0.05,spearman rank correlation coefficient is 0.989).3.100 patients had both pathology and HPV E6/E7 mRNA test results,effectiveness Analysis included shunt.CIN class II,CIN grade III and cervical cancer were classified as CIN 2+ in biopsy pathology.NILM and CIN I are called <CIN 2.HPV E6/E7 mRNA detection of CIN 2+ cervical lesions of the sensitivity of 82.69%,specificity of 62.50%,positive predictive value of 70.49%,negative predictive value of76.92%.4.100 patients have both pathology and TCT test results,include into the analysis of shunt efficiency.CIN class II,CIN grade III and cervical cancer are classified as CIN 2+ in biopsy pathology.NILM and CIN I are classified as <CIN 2.NILM ?ASC and LSIL in the TCT test results are TCT negative.While HSIL and SCC/AGC are classified as TCT positive.TCT detection of CIN 2+ cervical lesions of the sensitivity of 65.38%,specificity of 60.42%,positive predictive value of 64.15%,negative predictive value of 61.70%.5.100 patients with biopsy pathology,at the same time they have HPV E6/E7 mRNA and TCT test results,include into the analysis of shunt efficiency.CIN class II,CIN grade III and cervical cancer are classified as CIN 2+ in biopsy pathology.NILM and CIN I are called <CIN 2.The TCT results of HSIL ?SCC/AGC and positive results of HPV E6/E7 mRNA detection are all classified as positive for combined test.The TCT results of NILM ?ASC ?LSIL or negative results of HPV E6/E7 mRNA test are all classified as negative for combined detection.HPV E6/E7 mRNA combined TCT test of CIN 2+ cervical lesions of the sensitivity of 63.46%,specificity of 75.00%,positive predictive value of 73.33%,negative predictive value of65.45%.Conclusion:1.The expression of HPV E6/E7 mRNA was detected in patients with different pathological grades.Statistical analysis showed: With the degree of cervical lesions increased,HPV E6/E7 mRNA positive rate was on the rise,the positive rate of HPV E6/E7 mRNA was positively correlated with pathological grade.The expression of HPV E6/E7 mRNA in cervical tissues with different pathological grades was strongly correlated with pathological grade.2.HPV E6/E7 mRNA detection can be used for shunt patients with HPV-positive patients.To avoid the false positive of HPV DNA detection in clinical practice.3.HPV E6/E7 mRNA detection combined with TCT for cervical cancer sceening,help to predict the extent of disease progression,effectively shunt patients with pre-cervival lesions,effectively manage the population.