探讨人乳头瘤病毒(HPV)-E6/E7 mRNA检测，在宫颈癌初筛(宫颈癌初筛Cervista法检测)结果为高危型HPV DNA呈阳性受试者的临床价值。

选取2015年11月30日至2016年11月30日，在湖州市妇幼保健院妇科就诊的437例高危型HPV DNA呈阳性受试者为研究对象。按照年龄，将其分为≥25～30岁组(n＝118)、≥30～40岁组(n＝117)、≥40～50岁组(n＝129)、≥50～60岁组(n＝52)、≥60～65岁组(n＝21)。对所有受试者进行新柏液基薄层细胞学检查(TCT)和支链DNA(bDNA)技术检测宫颈脱落细胞HPV-E6/E7 mRNA表达情况及阴道镜检查，并对可疑部位取组织标本进行活组织病理学检查，以病理学检查结果为"金标准"。比较不同年龄组、不同组织病理学诊断结果受试者HPV-E6/E7 mRNA表达情况，评估HPV-E6/E7 mRNA检测在宫颈病变筛查中的价值。不同年龄组、不同组织病理学诊断结果受试者的HPV-E6/E7 mRNA阳性率比较，以及HPV-E6/E7 mRNA检测与TCT预测组织病理学诊断结果为宫颈病变≥高级别鳞状上皮内瘤变(HSIL)的敏感度、特异度、阳性预测值、阴性预测值比较，采用χ²检验或Fisher确切概率法。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求，并与所有受试者签署临床研究知情同意书。

①不同年龄组高危型HPV DNA呈阳性受试者的HPV-E6/E7 mRNA阳性率比较，差异无统计学意义(P>0.05)。②对437例高危型HPV DNA呈阳性受试者的组织病理学诊断结果显示，正常宫颈组织为149例，低级别鳞状上皮内瘤变(LSIL)为60例，HSIL为215例[宫颈上皮内瘤变(CIN)2、3级各为73例和142例]，宫颈鳞状细胞癌(SCC)为9例，宫颈腺癌为4例。③组织病理学诊断结果为宫颈病变≥HSIL受试者的HPV-E6/E7 mRNA阳性率为98.2%(224/228)，显著高于组织病理学诊断结果为宫颈病变≤LSIL者(45.5%，95/209)，并且差异有统计学意义(χ²＝151.507，P<0.001)。④在437例高危型HPV DNA呈阳性受试者中，HPV-E6/E7 mRNA检测对于预测组织病理学诊断结果为宫颈病变≥HSIL的敏感度和阴性预测值，均显著高于TCT，分别为98.2%(224/228) vs 71.1%(162/228)和96.6%(114/118) vs 68.7%(145/211)，并且差异均有统计学意义(χ²＝64.870、35.140，P<0.001)；其特异度显著低于TCT ，分别为54.6% (114/209)与69.4%(145/209)，并且差异亦有统计学意义(χ²＝8.925，P＝0.003)；而二者的阳性预测值比较，差异无统计学意义(χ²＝0.137，P＝0.711)。⑤在TCT结果分别为无上皮内病变或恶性病变(NILM)与未明确意义的非典型鳞状上皮细胞(ASC-US)的高危型HPV DNA呈阳性受试者中，HPV-E6/E7 mRNA检测，对于预测组织病理学诊断结果为宫颈病变≥HSIL的敏感度、特异度、阳性预测值、阴性预测值分别为97.0%(64/66)、56.6%(82/145)、50.4%(64/127)、97.6%(82/84)与100.0%(32/32)、62.5%(20/32)、72.7%(32/44)、100.0%(20/20)。

使用bDNA技术检测HPV-E6/E7 mRNA表达情况，对宫颈癌初筛结果为高危型HPV DNA呈阳性受试者的宫颈病变进一步筛查具有一定临床价值。采用bDNA技术对高危型HPV DNA呈阳性，而TCT结果为NILM或ASC-US受试者进行分流，可降低宫颈病变漏诊率和阴道镜转诊率。

To explore the clinical values of human papilloma virus (HPV)-E6/E7 mRNA detection in high-risk HPV DNA-positive women(detected by Cervista HPV).

From November 30, 2015 to November 30, 2016, a total of 437 high-risk HPV DNA-positive women were enrolled as eligible women from Huzhou Maternity & Child Care Hospital. According to age of them, they were divided into ≥25-30 year-old group (n=118), ≥30-40 year-old group (n=117), ≥40-50 year-old group (n=129), ≥50-60 year-old group (n=52), and ≥60-65 year-old group (n=21). All eligible women underwent ThinPrep liquid-based cytology test (TCT), HPV-E6/E7 mRNA expressions in cervical exfoliated cells by branched DNA (bDNA), and colposcopy. Histopathological examination of biopsies was conducted on suspicious tissues, and its results were set as the " gold standard" . Consistency of results between histopathological examination of biopsies and cytopathological examination cervical exfoliated cells were compared. Expressions of HPV-E6/E7 mRNA in groups with different ages and subjects with different results of histopathological diagnosis were compared and its values in cervical lesions screening were assessed. Chi-square test or Fisher exact test was used to compare the positive expression rates of HPV-E6/E7 mRNA among groups with different ages and subjects with different results of histopathological diagnosis, and to compare the sensitivities, specificities, positive predictive values and negative predictive values between HPV-E6/E7 mRNA test and TCT in predicting cervical lesion with high-grade squamous intraepithelial lesion (HSIL)+ (HSIL or worse) by histopathological diagnosis. This study met the requirements of the World Medical Association Declaration of Helsinki revised in 2013. Clinical informed consent was obtained from each subject.

①There was no significant difference among groups with different ages in positive expression rates of HPV-E6 /E7 mRNA in subjects with high-risk HPV DNA-positive (P>0.05). ②The histopathological results of 437 women with high-risk HPV DNA-positive were as following: 149 cases of normal cervix, 60 cases of low-grade squamous intraepithelial lesion (LSIL), 215 cases of HSIL, including 73 cases of cervical intraepithelial neoplasia (CIN) 2 and 142 cases of CIN3, 9 cases of squamous cell carcinoma (SCC) and 4 cases of adenocarcinoma. ③Positive expression rate of HPV-E6/E7 mRNA in HSIL+ subjects was significantly higher than that in LSIL- (LSIL or better) subjects which were 98.2% (224/228) and 45.5% (95/209), respectively, and the difference was statistically significant (χ²=151.507, P<0.001). ④Among 437 cases of HPV DNA-positive subjects, the sensitivity and negative predictive value of HPV-E6/E7 mRNA detection in predicting histopathological examination of HSIL+ were much higher than those of TCT, which were 98.2% (224/228) vs 71.1% (162/228), and 96.6% (114/118) vs 68.7% (145/211), respectively, and both the differences were statistically significant (χ²=64.870, 35.140, P<0.001), and its specificity was remarkably lower than that of TCT, which were 54.6% (114/209) vs 69.4% (145/209), respectively, and the difference was statistically significant (χ²=8.925, P=0.003); but there was no significant difference in positive predictive values between those two methods (χ²=0.137, P=0.711). ⑤Among the high-risk HPV DNA-positive women with no intraepithelial lesion or malignant (NILM) or atypical squamous cell of undetermined significance (ASC-US), the sensitivities, specificities, positive predictive values and negative predictive values of HPV-E6/E7 mRNA detection in predicting histopathological examination of HSIL+ were 97.0% (64/66), 56.6% (82/145), 50.4% (64/127), 97.6% (82/84), and 100.0% (32/32), 62.5% (20/32), 72.7% (32/44), 100.0% (20/20), respectively.

Detection of HPV-E6/E7 mRNA by bDNA technology has certain value for cervical lesions screening of high-risk HPV DNA-positive women, and can reduce the rate of missed diagnosis and colposcopy referral for high-risk HPV DNA-positive and cytological NILM or ASC-US women.