高危型人乳头状瘤病毒E6/E7mRNA检测对宫颈病变的诊断研究

目的探讨高危型人乳头状瘤病毒E6/E7 mRNA检测对宫颈病变的诊断价值。方法选择我院2019年3月-11月接诊的疑似宫颈病变患者,筛选163例患者,采用电脑随机分组的方式,163例分为81例对照组以及82例研究组,对照组接受高危型人乳头瘤病毒(human papollomavirus,HPV)DNA检测,研究组患者均接受高危型人乳头状瘤病毒E6/E7 mRNA检测,对比两组患者的检测准确率。结果研究组慢性宫颈炎、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)I型宫颈癌诊断准确率均高于对照组,有统计学意义(P<0.05);研究组与对照组在CIN Ⅱ型、CIN Ⅲ型诊断准确率上虽无统计学意义,但是研究组的准确率高于对照组。结论对于疑似宫颈病变的患者而言,接受高危型人乳头状瘤病毒E6/E7 mRNA检测,不仅能够有效的判断疾病的类型,且诊断准确率较高,值得推广。     

人乳头瘤病毒E6／E7mRNA检测在宫颈病变筛查中的应用价值

目的：探讨人乳头瘤病毒（HPV）E6/E7 mRNA 检测在宫颈病变筛查中的应用价值。方法2012年6月至2013年4月在余姚市人民医院因宫颈炎症、阴道异常出血或排液的720例门诊患者行宫颈 TCT 和 HPV DNA 检测,对 TCT 结果≥ASC-US 或（和）高危 HPV DNA 阳性者行宫颈 HPV E6/E7mRNA 检测和组织病理学检查。以病理学结果为标准,与 HPV DNA 检测相比较,评价 HPV E6/E7 mRNA 检测对 CIN2＋诊断价值,并用受试者工作特征（ROC）曲线评价其准确性。结果在 CIN 和宫颈浸润癌中,HPV E6/E7 mRNA 总体阳性率为78．7％,低于 HPV DNA 总体阳性率（92．6％）,差异有统计学意义（P ＜0．001）。HPV E6/E7 mRNA 检测诊断 CIN2＋的灵敏度为83．0％,低于 HPV DNA（94．5％）,差异有统计学意义（P ＜0．01）;特异度为51．1％,高于 HPV DNA（22．8％）,差异有统计学意义（P ＜0．001）。 HPV E6/E7 mRNA、HPV DNA 对诊断 CIN2＋的 ROC 曲线下面积分别为0．670、0.587, HPV E6/E7 mRNA 检测对 CIN2＋的诊断的准确性高于 HPV DNA,差异有统计学意义（P ＜0．01）。结论 HPV E6/E7 mRNA 检测诊断 CIN2＋特异度和准确性高于 HPV DNA,对宫颈病变的筛查有一定应用价值。     

Human papillomavirus testing for primary cervical cancer screening

Cervical cancer is the second most common cancer in women worldwide and the seventh most common cause of cancer deaths in women in Europe. Today, we know how to prevent almost every case of this disease; organized cervical cancer screening based on the Papanicolaou or Pap smear has been proven to prevent 80% of cervical cancer deaths, while new technologies for the detection of the human papillomavirus (HPV) or the prevention of HPV infection offer the potential to make even more progress in the battle against this disease. Testing for carcinogenic or high-risk HPV types is gaining acceptance for the triage of women with borderline cytology and for follow-up after treatment of high-grade cervical lesions. Now, a number of large-scale randomized controlled trials have shown that HPV testing as a primary screening test can detect approximately 50% more high-grade lesions than the Pap test, albeit with a lower specificity if all HPV-positive women are followed up. However, alternative screening algorithms in which HPV-positive women are triaged with cytology have been shown to have equivalent specificity to the Pap test without compromising the increased sensitivity. Further advantages of HPV testing come from the fact that it is an objective and automatable test with a dichotomous result. These attributes can yield cost savings through reductions in staff numbers and simplification of quality control procedures while reducing turnaround times. In countries seeking to improve cervical cancer prevention, the implementation of HPV testing as the primary screening test with cytology for the triage of HPV-positive women is an option that should be fully evaluated. This review summarizes the recent advances in HPV testing in cervical cancer prevention.     

应用支链DNA技术检测人乳头瘤病毒E6/E7mRNA在宫颈疾病筛查中的价值

目的评价支链DNA(bDNA)技术检测人乳头瘤病毒(HPV)E6/E7mRNA在宫颈疾病筛查中的临床价值。方法对262例宫颈疾病筛查妇女宫颈脱落细胞采用bDNA技术检测HPVE6/E7 mRNA、第二代杂交捕获技术(HC2)检测高危HPV,同时进行宫颈病理学检查。结果 (1)随着宫颈疾病级别升高,bDNA技术和HC2法检测HPV阳性率均呈现趋势性增加(χ2=65.397,P<0.001;χ2=65.018,P<0.001);(2)bDNA技术对总体人群检出阳性率为39.7%,HC2法检出阳性率为56.1%,HC2法显著高于bDNA技术(χ2=13.489,P<0.001),二者一致性一般(Kappa=0.501);(3)bDNA技术对CIN2+诊断的灵敏度为63.4%(95%CI:54.5%～72.3%),显著低于HC2法78.6%(95%CI:71.0%～86.2%)(χ2=5.549,P=0.018);而特异性为78.0%(95%CI:71.4%～84.6%),高于HC2的60.7%(95%CI:52.8%～68.5%)(χ2=9.798,P=0.002)。结论 HC2法检测高危HPV仍是目前宫颈疾病筛查...     

宫颈细胞HPV E6/E7 mRNA检测在宫颈病变诊断中的价值研究

目的探讨宫颈细胞人乳头状瘤病毒(HPV)E6/E7mRNA检测对于诊断宫颈病变的应用价值。方法选取2014年10月至2015年10月在该院妇产科因宫颈炎性反应、阴道异常出血或排液门诊患者就诊的407例患者,行新柏氏液基细胞学检测(TCT)和HPV E6/E7mRNA检测,对TCT结果异常的34例患者行病理科组织病理学检查,以病理报告为"金标准"。统计分析HPV E6/E7mRNA检测对宫颈病变的应用价值。结果 TCT检测出异常患者34例,占总患者数的8.35%,无上皮内瘤变或恶性病变(NILM)患者373例,占总患者数的91.65%;HPV E6/E7mRNA检测异常患者87例,占总患者数的21.37%。2组差异有统计学意义(P0.05)。TCT不同结果与HPV E6/E7mRNA拷贝数的比较表明,从意义不明的不典型鳞状细胞(ASCUS)到不能排除高度上皮内病变的不典型鳞状上皮(ASC-H)、鳞状上皮内低度病变(LSIL)、鳞状上皮内高度病变(HSIL),随着细胞学诊断级别的升高,HPV E6/E7mRNA阳性率同步升高(P0.05)。结论宫颈细胞HPV E6/E7mRNA诊断可较大程度降低过度检查和治疗的可能性,诊断宫颈上皮内瘤变(CIN)Ⅱ+的准确性较高,可降低宫颈病变的漏诊率,其对于宫颈病变的诊断有较高的临床应用价值。     

HPV E6／E7 mRNA 检测在葫芦岛地区宫颈癌筛查中的应用

目的：探讨人乳头瘤病毒（HPV）E6/E7 mRNA 检测在宫颈癌筛查中的应用。方法选取于该院妇科治疗的160例患者的超薄液基细胞学检查（thin prep cytology test,TCT）标本,根据患者的检查结果将其分为4个组：正常组（32例）、宫颈上皮内瘤变（CIN）Ⅰ组（44例）、CINⅡ组（67例）、CINⅢ组（17例）。除了 CIN 分级外,又根据不同的病理学类型对患者进行分组。在不同的组别中,比较 HPV E6/E7 mRNA 检测和 HPV DNA 检测阳性率的差异;并比较不同病理学分组间 HPV E6/E7 mRNA 和HPV DNA 水平的差异。结果在不同 CIN 分级的患者及对照组人群中,HPV E6/E7 mRNA 和 HPV-DNA 的阳性率比较,差异均有统计学意义（P ＜0．05）。对不同病理学分组间 HPV E6/E7 mRNA 和 HPV DNA 水平进行比较,差异有统计学意义（P ＜0．05）。结论在宫颈癌的筛查中 HPV E6/E7 mRNA 检测可以作为筛选手段之一,该项检测和细胞学检测联合运用有助于诊断准确性的提高。     

p16/Ki-67双染联合高危型人乳头瘤病毒E6/E7mRNA检测对子宫颈癌前病变诊断意义的初步评价

目的探讨p16/Ki-67双染联合高危型人乳头瘤病毒E6/E7mRNA检测对子宫颈癌前病变(CINⅡ+病变,包括CINⅡ、CINⅢ及宫颈原位癌)诊断的意义。方法应用宫颈炎标本176例、CINⅠ标本121例、CINⅡ标本95例、CINⅢ标本86例和宫颈原位癌标本75例为研究对象。回顾分析前期细胞学样本p16/Ki-67双染、HPV E6/E7mRNA检测结果,观察p16/Ki-67双染和HPV E6/E7mRNA检测在同级细胞学、组织学诊断中阳性率的差异。比较p16/Ki-67双染、HPV E6/E7mRNA、p16/Ki-67双染联合HPV E6/E7mRNA发现CINⅡ(+)病变的差异。结果在未见上皮内瘤变及恶性病变(NILM)组与意义不明的非典型鳞状细胞(ASCUS)组,p16/Ki-67双染和HPV E6/E7mRNA检测阳性率差异均显著(P<0. 05);而在低级别鳞状上皮内瘤变(LSIL)组与高级别鳞状上皮内瘤变以上组(HSIL+),差异不显著(P>0. 05)。②在宫颈炎、CINⅠ、CINⅡ、CINⅢ、宫颈原位癌组,p16/Ki-67双染阳性率和HPV E6/E7mRNA检测阳性率差异均不显著(P>0. 05)。③三种检测方法诊断CINⅡ(+)病变的灵敏度、特异度、符合率、阳性预测值、阴性预测值总体差异均显著(P<0. 05)。p16/Ki-67双染联合HPV E6/E7mRNA检测法诊断CINⅡ(+)病变的灵敏度、阴性预测值最高,分别为95. 70%、84. 85%、89. 87%、84. 48%、95. 82%。结论 p16/Ki-67双染、HPV E6/E7mRNA检测均可应用于宫颈癌前病变的筛查。相比较于单独检测,两者联合应用明显提高了CINⅡ(+)病变灵敏度、阴性预测值,提高了筛查方法的效率,有在临床推广使用的潜能。     

Human papillomavirus viral load: a possible marker for cervical disease in HIV-infected women

Laboratory markers of human papillomavirus infection have been recognized as relevant tools in programmes designed to reduce the burden of cervical cancer. The ongoing experience with these laboratory markers serves to confirm not only their negative predictive value (close to 100%) but also their positive association with developing or developed lesions. This aspect is particularly relevant in HIV-infected subjects who show an increased prevalence, incidence and severity of infections and lesions even in the era of efficacious control of their immunosuppression. Among the possible virus-related parameters proposed as relevant markers (viral persistence, load, expression, genomic integration capacity) we here analyse the informative value of human papillomavirus viral load measurement as a possible risk marker in this particular clinical setting.     

A therapy modality using recombinant IL-12 adenovirus plus E7 protein in a human papillomavirus 16 E6/E7-associated cervical cancer animal model

Interleukin (IL)-12 has been reported to induce cellular immune responses for protection against tumor formation. Here we investigate the utility of adenoviral delivery of IL-12 as an adjuvant for a human papillomavirus E7 subunit vaccine in a mouse tumor challenge model. Direct intratumoral injection of AdIL-12 resulted in a significant suppression of tumor growth compared to the control group. Injection of E7 protein into either a tumor site or the distance site along with AdIL-12 further enhanced antitumor effects significantly higher than either AdIL-12 or E7 injection alone. This combined injection resulted in complete regression of 9-mm-sized tumor in 40% of animals as well as lasting antitumor immunity against tumor recurrence. We also evaluated immune responses induced by these injections. AdIL-12 plus E7 enhanced E7-specific antibody responses significantly higher than AdIL-12 or E7 injection. In particular, the production level of interferon (IFN)-gamma from E7-specific CD4(+) T cells was similar between AdIL-12 group and AdIL-12 + E7 group. However, IFN-gamma production from E7-specific CD8(+) T cells was the most significant when injected with AdIL-12 + E7. This was consistent with intracellular IFN-gamma staining levels of CD8(+) T cells, suggesting that AdIL-12 + E7 injection enhances antitumor immunity in the human papillomavirus (HPV) 16 tumor model through increased expansion of the cytotoxic T-lymphocyte (CTL) subset. This enhanced protection appeared to be mediated by CD8(+) T cells, as determined by in vivo T-cell subset deletion. Thus, these studies demonstrate that E7 vaccines can induce CTL responses responsible for antitumor effects in the presence of IL-12 delivered via adenovirus vectors. This likely provides one additional approach for immune therapy against cervical cancers.     

多重实时荧光定量PCR方法在高危型人乳头瘤病毒E6/E7 mRNA检测中的应用

摘要：目的?建立一种基于多重实时荧光定量PCR(MRT-PCR)技术的可同时检测14种高危亚型人乳头瘤病毒(HPV16、18、31、33、35、39、52、45、51、56、58、59、66、68)癌基因E6/E7 mRNA的方法。方法?对14种高危型HPV各自的E6/E7基因序列区域设计特异性引物和探针，配以优化的反应体系，形成HPV E6/E7 mRNA检测体系，评价方法检出限、特异性。收集223例临床样本，分别用自建的一步法MRT-PCR法与转录介导等温核酸扩增(TMA)技术(Aptima?HPV试剂盒)检测HPV E6/E7 mRNA，同时评估HPV E6/E7 mRNA检测结果与薄层液基细胞学检测、宫颈组织病理学检测结果的一致性。结果?建立的检测方法对常见的低危型HPV无交叉检出，且对14种高危型HPV的检测限可达10～100 copies/μL。该方法检测223例临床标本的结果与Aptima??HPV检测结果相比，一致性较好(Kappa=0.910)。以组织病理结果为CINⅡ及以上的作为阳性，MRT-PCR法临床检测敏感性为84.0%，特异性为94.4%，阳性预测值为65.6%，阴性预测值为97.9%。结论?建立的MRT-PCR方法具有特异性好、灵敏度高和稳定性好等优点，为HPV临床快速检测以及宫颈病变筛查提供了一个有效的技术平台。     

HPV DNA和HPV E6/E7 mRNA检测在宫颈癌筛查中的应用价值探讨

目的探讨HPV DNA和HPV E6/E7 mRNA检测在宫颈癌筛查中的应用价值。方法选取宫颈病变筛查的受试者407例为研究对象,均行宫颈薄层液基细胞学(TCT)检查,并检测细胞学标本中HPV DNA和HPV E6/E7 mRNA的表达。215例行阴道镜宫颈活检,结合组织病理诊断结果进行统计分析。结果 407例细胞病理学诊断结果:正常范围(WNL)192例(47.17%)、意义不明的不典型鳞状上皮细胞(ASCUS)106例(26.04%)、低度鳞状上皮内病变(LSIL)70例(17.20%)和高度鳞状上皮内病变(HSIL)39例(9.58%)。诊断为LSIL和HSIL患者中,HPV DNA和HPV E6/E7 mRNA检出率差异无统计学意义(P0.05);诊断为ASCUS患者中,HPV DNA检出率显著高于HPV E6/E7 mRNA(P0.01)。215例组织病理学诊断结果:正常109例,阴性106例,包括CINⅠ级44例、CINⅡ级27例、CINⅢ级32例、宫颈鳞癌3例。对于诊断为CINⅡ级、CINⅢ级/宫颈癌患者,HPV DNA和HPV E6/E7 mRNA检测的灵敏度、特异度和阳性预测值差异无统计学意义(P0.05),但HPV E6/E7 mRNA检测的阴性预测值和准确度显著高于HPV DNA检测(P0.05或P0.01)。结论 HPV E6/E7 mRNA检查较HPV DNA检查能更好地评估HPV感染患者宫颈病变恶化风险,联合TCT检测对提高宫颈癌早期筛查和诊断的准确度具有积极意义。     

惠州地区高危型HPVE6/E7 mRNA检测在宫颈癌筛查中的临床意义

目的探讨与液基细胞学(TCT)检测结果比较,分析高危型人乳头瘤病毒(HPV)E6/E7 mRNA检测在宫颈癌筛查中的临床价值。方法选取该院妇科接受TCT检查的140例患者,根据检查结果分为5个组:炎性/良性组(32例),宫颈上皮内瘤变(CIN)Ⅰ组(44例),INⅡ组(38例),CINⅢ组(17例),宫颈癌组(9例)。比较各组HPV E6/E7mRNA检测阳性率和表达水平。结果 CIN异型程度增高,HPV E6/E7 mRNA阳性率和表达水平均呈增加趋势,不同CIN分级患者的阳性率和表达水平比较,差异均有统计学意义(P0.05);与TCT病理检查结果有较高的一致性。结论高危型HPV E6/E7 mRNA可以作为惠州地区宫颈癌筛查手段之一,联合HPV E6/E7 mRNA及TCT检测可提高其诊断准确性。     

人乳头瘤病毒E6／E7mRNA在不同程度宫颈病变中的应用价值

目的：探讨人乳头瘤病毒（HPV）E6／E7 mRNA水平在不同程度宫颈病变中的应用价值。方法选取2011年4月至2013年4月广东省中医院妇科门诊收治的HPV感染患者430例,将HPV‐DNA阳性标本根据病理资料分为炎症组、CINⅠ组、CIN Ⅱ组、CIN Ⅲ组和宫颈癌组。用反转录聚合酶链反应（RT‐PCR）检测各组标本的HPV E6／E7 mRNA水平,比较不同宫颈病变组间HPV‐DNA病毒载量及 HPV E6／E7 mRNA阳性检出率的差异情况。结果随着宫颈病变程度增加,HPV‐DNA病毒载量未呈现逐级递增的趋势,各病变组间的病毒载量两两比较差异无统计学意义（P＞0．05）;而HPV E6／E7 mRNA阳性检出率则逐级递增,除炎症组与CINⅠ组的检出率比较差异无统计学意义（P＞0．05）之外,其余各组之间的检出率两两比较差异均有统计学意义（P＜0．05）。结论 HPV E6／E7 mRNA与宫颈病变程度有高度的相关性,可作为宫颈疾病筛查的重要检测指标。     

Genetic immunization against cervical carcinoma: induction of cytotoxic T lymphocyte activity with a recombinant alphavirus vector expressing human papillomavirus type 16 E6 and E7

Infection of genital epithelial cells with human papillomavirus (HPV) types 16 and 18 is closely associated with the development of cervical carcinoma. The transforming potential of these high-risk HPVs depends on the expression of the E6 and E7 early viral gene products. Since the expression of E6 and E7 is selectively maintained in premalignant and malignant cervical lesions these proteins are attractive candidates for immunotherapeutic and prophylactic strategies. This report describes the construction, characterization and the in vivo immunotherapeutic potential of recombinant Semliki Forest virus (SFV) expressing the HPV16 E6 and E7 proteins (SFV-E6E7). Western blot analysis and immunofluorescence staining demonstrated expression of E6 and E7 in BHK cells infected with SFV-E6E7. Immunization of mice with SFV-E6E7 resulted in an efficient in vivo priming of HPV-specific CTL activity. The induced CTL lysed murine tumor cells transformed with the HPV16 genome and EL4 cells loaded with an immunodominant class I-binding HPV E7 peptide. CTLs could reproducibly be induced by immunization with three injections of as few as 10(5) infectious units of SFV-E6E7. Protection from tumor challenge was studied using the tumor cell line TC-1. Immunization with 5 x 10(6) SFV-E6E7 particles protected 40% of the mice from tumor challenge. These results indicate that E6E7 expression by the efficient and safe recombinant SFV system represents a promising strategy for immunotherapy or immunoprophylaxis of cervical carcinoma.     

Aptima HPV E6/E7 mRNA 16,18/45基因型检测技术在宫颈癌前病变和宫颈癌筛查中的应用价值

目的 探讨人乳头瘤病毒(HPV)E6/E7 mRNA 16,18/48基因型检测在宫颈癌筛查中的应用价值.方法 收集福建省宁德市闽东医院2015-2019年间Aptima HPV E6/E7 mRNA定性检测阳性患者871例,进行Aptima HPV E6/E7 mRNA 16、18/45基因型检测,追踪其宫颈液基细胞...     

FRD联合HR-HPV DNA、HPV E6/E7 mRNA检测对宫颈上皮内瘤变的诊断意义

目的探讨叶酸受体介导的宫颈特殊染色法(FRD)联合高危人乳头状瘤病毒(HR-HPV)DNA、人乳头状瘤病毒(HPV)E6/E7 mRNA检测对宫颈上皮内瘤变(CIN)的诊断意义。方法以2018年5月至2020年4月佛山市第一人民医院进行诊治的200例CIN患者作为观察组,另选取同期100例健康妇女作为对照组。对所有受试者进行FRD、HR-HPV DNA、HPV E6/E7 mRNA检测,比较2组受试者及不同病理分级CIN患者的检测结果。采用Spearman相关分析CIN诊断、病理分级与FRD、HR-HPV DNA、HPV E6/E7 mRNA检测阳性率的相关性,采用受试者工作特征(ROC)曲线评价FRD、HR-HPV DNA、HPV E6/E7 mRNA单独和联合检测对CIN的诊断效能。结果FRD检测CIN的假阳性率为3.00%,假阴性率为8.00%,2组FRD诊断CIN的准确率差异无统计学意义(P>0.05);CINⅡ级、Ⅲ级患者FRD检测的阳性率均高于CINⅠ级(P<0.05)。2组HR-HPV DNA、HPV E6/E7 mRNA检测诊断CIN准确率比较,差异均有统计学意义(P<0.05);CINⅡ级、Ⅲ级患者HR-HPV DNA、HPV E6/E7 mRNA检测的阳性率均高于CINⅠ级(P<0.05)。CIN诊断、病理分级与FRD、HR-HPV DNA、HPV E6/E7 mRNA检测的阳性率均呈正相关(r>0,P<0.05)。FRD、HR-HPVDNA、HPVE6/E7mRNA联合检测诊断CIN的曲线下面积(AUC)、灵敏度和特异度均明显高于单独检测诊断的AUC、灵敏度和特异度(P<0.05)。结论FRD、HR-HPV DNA、HPV E6/E7 mRNA联合检测对CIN具有较高的诊断效能,值得临床推广。     

Human papillomavirus vaccines for cervical cancer.

Cervical cancer is one of the most common causes of cancer-related death in women. As a result of several recent advances in molecular biology, the association between human papillomavirus (HPV) infection and cervical cancer has been firmly established, and the oncogenic potential of certain HPV types has been clearly demonstrated. Several lines of evidence suggest the importance of the host's immune response, especially cellular immune response, in the pathogenesis of HPV-associated cervical lesions. These observations form a compelling rationale for the development of vaccine therapy to combat HPV infection. Both prophylactic and therapeutic HPV vaccine strategies are being developed. Prophylactic strategies currently under investigation focus on the induction of effective humoral immune responses against subsequent HPV infection. In this respect, impressive immunoprophylactic effects have been demonstrated in animals using papillomavirus-like particles (VLPs). VLPs are antigenic and protective, but are devoid of any viral DNA that may be carcinogenic to the host. For treatment of existing HPV infection, techniques to improve cellular immunity by enhancing viral antigen recognition are being studied. For this purpose, the oncogenic proteins E6 and E7 of HPV-16 and -18 are the focus of current clinical trials for cervical cancer patients. The development of successful HPV-specific vaccines may offer an attractive alternative to existing screening and treatment programs for cervical cancer.