HPV E6/E7 mRNA检测在HPV分型及宫颈疾病筛查中的作用研究

Clinical study of HPV E6/E7 mRNA detection in HPV typing and screening of cervical diseases

目的探讨人乳头瘤病毒(HPV)E6/E7 m RNA检测在女性宫颈疾病筛查中的作用。方法选取2016年9月至2017年9月贵州医科大学附属医院妇产科门诊和住院部就诊的HPV阳性感染者287例,其中宫颈疾病患者267例[包括宫颈炎、宫颈上皮内瘤变(CIN)Ⅰ~Ⅲ、宫颈癌],阳性感染者中体检无疾病表现及病理学检查无宫颈病变者20例。取患者宫颈脱落细胞用支链DNA技术(简称b-DNA技术)检测高危HPV E6/E7 m RNA,患者同时行宫颈液基薄层细胞检测和HPV分型检测,对不同年龄、HPV分型、阴道细胞学分型(简称TBS分型)及疾病分级的HPV E6/E7 m RNA的诊断价值进行效能分析。结果在不同年龄段内E6/E7阳性和阴性所占比例差异有统计学意义,P<0.05,在年龄组≤30岁组和>50~60岁组,E6/E7阳性者所占比例最大,分别为85.40%和84.40%;而HPV E6/E7m RNA拷贝数在不同年龄组内差异没有统计学意义。E6/E7结果在单一感染和混合感染,以及单一感染和多重感染间差异无统计学意义;在不同感染型别中,只有HPV 16型随病变严重程度的增加其感染率呈递增趋势(P<0.05),其余HPV型与病变严重程度无明显相关性(P>0.05)。不同疾病分组间,E6/E7阳性率的分布差异有统计学意义,表现为无宫颈病变组(0)<宫颈炎(15.60%)<CINⅠ(86.20%)<CINⅡ(93.00%)<CINⅢ(96.50%)<宫颈癌(100.00%),P<0.05;E6/E7拷贝数表达差异有统计学意义,表现为宫颈炎1773.89(936.54)<CINⅠ4997.74(10 439.75)<CINⅡ52 655.02(38 495.10)<CINⅢ72 300.12(38 298.57)<宫颈癌121 616.68(34 348.59),P<0.05。结论在宫颈疾病中检测高危型HPVE6/E7 m RNA的转录对女性宫颈癌高危人群的筛查和诊治具有重要的价值,或可作为进一步分流HPV阳性病人的方法。

Objective To explore the detection of human papillomavirus E6/E7 mRNA in the screening of women＇s cervical disease.Methods A total of 287 HPV-positive patients were selected from September 2016 to September 2017 in the Department of Obstetrics and Gynecology of the Affiliated Hospital of Guizhou Medical University,among whom 267 patients had cervical disease（including cervicitis,CINⅠ-CINⅢ and cervical cancer）.Another 287 HPV-postitive patients were chosen,among whom 20 had no disease manifestations on physical examination or cervical lesions on pathological examination.The branched-chain DNA technique（b-DNA）was used to detect the high-risk HPV E6/E7 mRNA in cervical exfoliated cells.Liquid-based thin-layer cervical cytology and HPV genotyping assays were used to analyze the diagnostic value of HPV E6/E7 mRNA in different ages,HPV typing,vaginal cytology type（TBS type）,and disease grading.Results The difference in the proportion of positive and negative E6/E7 in different age groups was statistically significant（P〈0.05）;the proportion of E6/E7-positive patients in the age group≤30 years and 〉50-60 years was the largest,being 85.40% and 84.40%,respectively.However,HPV E6/E7 mRNA copy number did not differ significantly among different age groups.The E6/E7 results also showed that there was no significant difference between single infection and mixed infection,or between single infection and multiple infections.Only HPV 16 showed an increasing rate of infection with the increasing severity of lesions（P〈0.05）,and there was no significant correlation between other HPV types and lesion severity in different infection types（P〉0.05）.The distribution of E6/E7-positive rates had statistical difference among different disease groups：no cercical lesions（0）〈cervicitis（15.60%）〈CIN Ⅰ（86.20%）〈 CINⅡ（93.00%）〈CINⅢ（96.50%） cervical cancer（100.00%）,P〈0.05.There was statistical difference in E6/E7 copy number,which was as follows：cervicitis 1773.89（936.54） 〈CINⅠ4997.74（10 439.75） 〈CINⅡ52 655.02（38 495.10）〈 CIN Ⅲ72 300.12（38 298.57） 〈cervical cancer 121 616.68（34 348.59）,P〈0.05.Conclusion Detection of high risk HPVE6/E7 mRNA transcription in cervical diseases is of great value to the screening and diagnosis of women at high risk of cervical cancer,and it can be used as a way to further divert HPV positive patients.