cobas 4800高危型人乳头瘤病毒检测技术介绍

正0引言子宫颈癌是常见妇科恶性肿瘤之一,全球发病率在女性恶性肿瘤中居第二位。子宫颈癌的发生是一个连续的过程,即由细胞分化失调到不典型增生,到原位癌,最后发展成子宫颈癌。目前研究发现,人乳头瘤病毒(HPV)是子宫颈癌及其前身宫颈上皮内瘤变(cervical intraepithelial neoplasias,CIN)发生的主要原因,超过99%的宫颈癌患者体     

人乳头瘤状病毒DNA载量与子宫颈癌前病变的关系

目的:研究妇女生殖道感染高危型人乳头瘤状病毒(HR-HPV)的载量与子宫颈上皮内瘤变(CIN)的关系。方法:病例来自我院2005年4月至2005年12月的住院病人,收集其子宫颈脱落细胞,采用第二代杂交捕获实验(HC-II)进行HPV DNA检测,病毒载量由样本的相对光单位(RLU)与标准阳性对照(PC)之比(RLU/PC)来衡量,按照log10RLU/PC分为阴性(0)、低度载量(0~1.14)、中度载量(1.15~2.28)和高度载量(2.29~3.44)。子宫颈病变按照病理诊断分为正常、CIN1、CIN2和CIN3。采用非条件多项式logistic回归分析病毒载量与子宫内瘤变(CIN)的关系。结果:98.0%(50/51)的CIN3、93.3%(28/30)的CIN2、72.2%(13/18)的CIN1、50%(161/322)的正常组HR-HPV DNA检测阳性,各组阳性对象的中位log10RLU/PC分别为2.18、1.59、1.49和1.47。从低度病毒载量与CIN1的相对危险度(OR)为1.7(0.5~7.2)至高度病毒载量与CIN3的OR值为88.6(11.6~676.7),显示出感染病毒载量与子宫颈病变程度呈正相关(P<0.001)。结论:子宫颈HR-HPV病毒载量是影响子宫颈癌前病变的重要危险因素。     

重庆市627例宫颈癌患者人乳头瘤病毒感染状况分析

目的：分析来自重庆市的宫颈癌患者的人乳头瘤病毒（human papilloma virus, HPV）感染状况及基因亚型分布,为宫颈癌的区域性防治提供参考。方法：对我院 2021 年 7 月至 12 月确诊的 627 例重庆籍宫颈癌患者进行 HPV 基因分型检测,并对检测结果进行统计分析。结果：宫颈癌患者的 HPV 感染率为 92.34%。以单一感染为主（71.61%）,二重感染次之（15.47%）。宫颈鳞癌的 HPV 总体感染率、二重感染率、多重感染率均明显高于宫颈腺癌,差异有统计学意义（P < 0.05）。总体检出率最高的前 5 位 HPV 亚型全部为高危人乳头瘤病毒,分别为：HPV16、18、58、52、33,感染率分别为 54.07%、7.66%、6.70%、4.47%、3.83%。不同年龄组宫颈癌患者 HPV 感染率之间,差异无统计学意义（χ2 = 1.705,P = 0.790）。HPV16 亚型作为优势亚型在不同年龄组宫颈癌患者中的感染率均为第 1 位,HPV52 亚型在 21 ~ 29 岁年龄组感染率较高,HPV58 亚型在 40 ~ 59 岁年龄组感染率较高,HPV18 亚型癌 30 ~ 39 岁和≥ 60 岁年龄组的感染率较高。结论：重庆地区宫颈癌患者 HPV 感染以单一感染为主,二重感染次之;感染率最高的前 5 位 HPV 亚型分别为：HPV16、18、58、52、33,全部为 HPV 高危亚型,且 HPV16亚型的感染率远远高于其他亚型。     

人乳头瘤病毒检测的研究进展

近年来,生殖道人乳头状病毒（HPV）的感染率日益增加,其临床与亚临床感染已成为当今社会流行最广泛的性传播疾病之一。文献报道,细胞学正常的妇女宫颈HPV感染率为10．2％～40．0％。HPV感染也是CIN、宫颈癌的明确病因,99％以上的宫颈癌有HPV感染。HPV检测广泛用于HPV相关恶性病变的自然史和病因学研究,目前它正在成为宫颈癌的另一种或是附加的筛检方法,对宫颈癌的预防及治疗有着非常重要的意义。     

人乳头瘤病毒检测与食管癌的研究进展

食管癌是我国常见的十大恶性肿瘤之一,越来越多的生物学和流行病学证据表明食管癌的发生与人乳头瘤病毒(HPV)感染有关,对食管癌中HPV的检测和型别研究,不仅可对食管癌病因学提供新的线索,而且对预防和治疗也具有积极意义。     

人乳头瘤病毒检测与食管癌的研究进展

食管癌是我国常见的十大恶性肿瘤之一,越来越多的生物学和流行病学证据表明食管癌的发生与人乳头瘤病毒（HPV）感染有关,对食管癌中HPV的检测和型别研究,不仅可对食管癌病因学提供新的线索,而且对预防和治疗也具有积极意义。     

cobas 4800与careHPV用于人乳头瘤病毒检测的对比研究

[目的]比较cobas 4800检测和careHPV人乳头瘤病毒（human papillomavirus,HPV）检测方法用于宫颈癌筛查的有效性和一致性。[方法]本研究纳入856名研究对象,采用cobas4800和careHPV检测方法对宫颈脱落细胞标本进行HPV DNA检测。以病理组织学为金标准,评估careHPV和cobas 4800检出宫颈上皮内瘤变2（cervical intraepithelial neoplasia grade 2,CIN2）及以上病人的有效性和准确性。采用McNemar检验对两种检测方法进行一致性检验。[结果]最终853例妇女的有效检测结果纳入统计分析。careHPV和cobas 4800检出HPV DNA阳性率分别是37.1%（316/853）和39.3%（335/853）。cobas 4800检测和careHPV检出HPV DNA一致率为83.2%（710/853）,Kappa=0.65（95%CI：0.59-0.70）。careHPV和cobas 4800检出CIN2＋的敏感度和特异性分别为94.4%（95%CI：81.3%-99.2%）和65.5%（95%CI：62.1%-68.7%）,94.4%（95%CI：81.3%-99.2%）和63.2%（95%CI：59.7%-66.5%）。在最后纳入统计分析的853名妇女中HPV16和HPV 18的感染率分别为13.1%和2.6%,而在36例CIN2＋的病人中,HPV16和HPV18的感染率分别为77.8%和2.8%。[结论]careHPV和cobas4800作为初筛方法检出CIN2＋的一致性较好,然而两种方法又有其不同的优势,应该根据当地的经济水平来选择用于筛查的HPV DNA检测方法。     

人乳头瘤病毒与宫颈癌关系的研究进展

人乳头瘤病毒(HPV)是能诱发宫颈癌的DNA病毒,尤其是高危型和混合感染.HPV的生物特性导致其具有致癌性,利用HPV的核酸检测可简便迅速地诊断宫颈癌,有利于早期治疗.鉴于HPV与宫颈癌的关系,HPV疫苗研制有重要意义.     

人乳头瘤病毒与泌尿系统肿瘤

人乳头瘤病毒与泌尿系统肿瘤江西医学院第一附属医院（３３０００６）徐鹏程综述王道仁审校近年来，随着肿瘤病毒病因研究的不断深入，人乳头瘤病毒（ＨＰＶ）与肿瘤的关系受到普遍关注，国外有关ＨＰＶ与膀胱癌、前列腺癌、尿道癌、阴茎癌关系的报道日益增多（１～８），...     

Comparison of human papillomavirus genotyping and cytology triage,COMPACT Study: Design,methods and baseline results in 14 642 women

Although cytology‐based screening programs have significantly reduced mortality and morbidity from cervical cancer, the global consensus is that primary human papillomavirus (HPV) testing for cervical screening increases detection of high‐grade cervical intraepithelial neoplasia (CIN) and invasive cancer. However, the optimal triage strategy for HPV‐positive women to avoid over‐referral to colposcopy may be setting specific. As Japan requires data that have been generated domestically to modify screening guidelines, we conducted a 3‐year prospective study, COMparison of HPV genotyping And Cytology Triage (COMPACT), to evaluate the potential role of HPV16/18 partial genotyping and cytology for primary HPV screening. In total, 14 642 women aged 20 to 69 years undergoing routine screening at 3 centers in Hokkaido were enrolled. Conventional cytology and HPV testing were carried out. Women with abnormal cytology or HPV16/18 positivity underwent colposcopy. Those with 12 other high‐risk (hr) HPV types underwent repeat cytology after 6 months. Primary study endpoints were detection of high‐grade cervical disease defined as CIN2/CIN3 or greater as determined by consensus pathology. Prevalence of cytological abnormalities was 2.4%. hrHPV, HPV 16, and HPV 18 were detected in 4.6%, 0.9%, and 0.3% of women, respectively. HPV16/18 were detected in all (8/8) invasive cervical cancers and in all (2/2) adenocarcinomas in situ. Both cytological abnormalities and hrHPV positivity declined with increasing age. This is the first Japanese study to investigate the role of partial genotyping and cytology in an HPV‐based screening program. Results should help policy‐makers develop guidelines for future cervical screening programs and management of cervical abnormalities based on HPV genotype.     

Distribution and incidence of atypical glandular lesions in cervical cytology focusing on the association with high-risk human papillomavirus subtypes

Adenocarcinoma in situ (AIS) is considered a precursor of adenocarcinoma. Cervical adenocarcinoma has been associated with human papillomavirus (HPV), while other subtypes of AIS and endocervical adenocarcinoma have no precursor lesions and are not associated with HPV. Cervical cytology and HPV genotyping are important in the detection of these different subtypes. Notably, endometrial lesions and infiltration with secondary adenocarcinoma may lead to misdiagnosis of endocervical lesions. The aim of the present study was to avoid misdiagnosis of squamous cell changes and endometrial lesions as endocervical lesions in cervical screening. A total of 210,510 female cytological samples were analyzed between the beginning of January 2020 and the beginning of January 2021. The samples were processed for conventional cytological techniques, and for molecular detection and subtyping of high-risk HPV (HPV-HR) according to the advice and measurements of BD Biosciences (117,765 samples) and the PapilloCheck^® HPV test (5,579 samples). The present study was carried out in Germany using the Munich classification III. II-g (Bethesda classification: atypical glandular cells not otherwise specified) was detected in 0.12% of cases under the age of 35 years. Another peak was noticed within the 41–60-years age group (0.11%). In the 41–50-years age group, a peak for II-e (Bethesda classification: Endometrial cells) (1.5%) was identified. An association was revealed between HPV16, HPV18 and HPV45 with cervical intraepithelial neoplasia III, AIS, endocervical adenocarcinoma and squamous cell carcinoma, in addition to other HPV-HR subtypes, such as HPV33/58, as well as 52, 56/59/66 in the different age groups. In patients aged <35 years, 0.03% of cases were vaccinated cases against HPV. In the 35–40-years age group, there was only one vaccinated case (0.0045%); in the 41–50-years age group, there were 11 vaccinated cases (0.031%); and in the 51–60-years age group, there was one vaccinated case (0.002%). No patients aged >60 years were vaccinated against HPV in the analyzed cohort. In conclusion, most cases of HPV-associated glandular dysplastic changes and neoplasia occurred in sexually active women aged between 35 and 60 years. In addition, endocervical adenocarcinoma may occur at any age with or without an HPV infection.     

Assessing the time dependence of prognostic values of cytology and human papillomavirus testing in cervical cancer screening

Accurate assessment of risks for developing cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) after a given set of screening test results is instrumental to reaching valid conclusions and informing cervical cancer screening recommendations. Using data from the Canadian Cervical Cancer Screening Trial (CCCaST), we assessed prognostic values of enrollment screening test results to predict CIN2+ among women attending routine cervical screening using multivariable Cox proportional hazards (PH) regression and its flexible extension during each of two follow-up periods (protocol-defined and extended). Nonproportional (time-dependent (TD)) and/or nonlinear effects were modeled, as appropriate. Women with abnormal cytology had hazard ratios (HRs) for CIN2+ detection of 17.61 (95% CI: 11.25–27.57) and 10.46 (95% CI: 5.41–20.24) relative to women with normal cytology during the protocol-defined and extended follow-up periods, respectively. High-risk human papillomavirus (HR-HPV) positivity was an even stronger predictor of CIN2+ risk, with significant TD effects during both follow-up periods (p <0.001 for both TD effects). Risks among women co-testing HR-HPV+ with and without abnormal cytology (relative to women co-testing negative) were highest immediately after baseline, and decreased significantly thereafter (p <0.001 for both TD effects). HRs for HPV16+ and HPV18+ women (relative to those testing HR-HPV-) did not vary significantly over time (HR = 182.96; 95% CI: 95.16–351.77 and HR = 111.81; 95% CI: 44.60–280.31, respectively). Due to TD effects, conventional Cox model estimates considerably underestimated adjusted HRs associated with positive HR-HPV testing results early on in the follow-up periods.     

人乳头瘤病毒检测阴性子宫颈癌的研究进展

子宫颈癌是妇科最常见的恶性肿瘤,研究证实99.7%的宫颈癌是因感染人乳头瘤病毒（human papillomavirus,HPV）造成的,但几乎所有的研究中都发现有HPV检测阴性的子宫颈癌存在。HPV检测阴性的子宫颈癌可以概括为假阴性和真阴性两种情况,造成假阴性的原因有病变小、取材有限、病毒载量低、非高危人乳头瘤病毒基因型、技术错误或检测灵敏度不足等。现有的筛查方式具有一定局限性,一些具有高灵敏度和特异性的新型分子标记物如微小核糖核酸、FAM19A4基因甲基化等已被证明有望作为子宫颈癌早期检测和诊断的指标。近年来关于HPV阴性子宫颈癌的研究越来越多,但对HPV阴性子宫颈癌患者临床特点的分析存在差异。本文主要对HPV阴性子宫颈癌假阴性的原因、筛查诊断及临床特点方面进行综述。     

A DNA methylation classifier of cervical precancer based on human papillomavirus and human genes

Testing for high‐risk (hr) types of human papillomavirus (HPV) is highly sensitive as a screening test of high‐grade cervical intraepithelial neoplastic (CIN2/3) disease, the precursor of cervical cancer. However, it has a relatively low specificity. Our objective was to develop a prediction rule with a higher specificity, using combinations of human and HPV DNA methylation. Exfoliated cervical specimens from colposcopy‐referral cohorts in London were analyzed for DNA methylation levels by pyrosequencing in the L1 and L2 regions of HPV16, HPV18, HPV31 and human genes EPB41L3, DPYS and MAL. Samples from 1,493 hrHPV‐positive women were assessed and of these 556 were found to have CIN2/3 at biopsy; 556 tested positive for HPV16 (323 CIN2/3), 201 for HPV18 (73 CIN2/3) and 202 for HPV31 (98 CIN2/3). The prediction rule included EPB41L3 and HPV and had area under curve 0.80 (95% CI 0.78–0.82). For 90% sensitivity, specificity was 36% (33–40) and positive predictive value (PPV) was 46% (43–48). By HPV type, 90% sensitivity corresponded to the following specificities and PPV, respectively: HPV16, 38% (32–45) and 67% (63–71); HPV18, 53% (45–62) and 52% (45–59); HPV31, 39% (31–49) and 58% (51–65); HPV16, 18 or 31, 44% (40–49) and 62% (59–65) and other hrHPV 17% (14–21) and 21% (18–24). We conclude that a methylation assay in hrHPV‐positive women might improve PPV with minimal sensitivity loss.     

维、汉族妇女宫颈鳞癌组织中人乳头瘤病毒基因分型研究

目的：了解维、汉族妇女宫颈鳞癌组织中人乳头瘤病毒（HPV）感染及其基因型的分布情况,并进一步探讨HPV感染及其基因型在维、汉族妇女宫颈鳞癌之间的分布是否有差异。方法：采用可检测23种HPV基因型的基因芯片方法分别检测30例维吾尔族宫颈鳞癌组织及30例汉族宫颈鳞癌组织的HPV基因型。结果：60例宫颈鳞癌组织中HPV的阳性率为95%（57/60）,其中维吾尔族宫颈鳞癌HPV的阳性率为96.6%（29/30）,汉族宫颈鳞癌HPV的阳性率为93.3%（28/30）,差异无统计学意义（P〉0.05）。60例宫颈鳞癌组织中共检测到6种HPV基因型,分别为HPV16,56,58,18,68,35,均为高危型感染;其中HPV16最常见,检出率为91.7%（55/60）,其次是HPV56,21.7%（13/60）;另外,60例宫颈鳞癌中共检测到HPV双基因型重复感染22例,检出率36.7%（22/60）。结论：HPV感染与维、汉族妇女宫颈鳞癌关系密切,尤其以HPV16型最有关,其次为HPV56型,并存在一定数量的多亚型重复感染。HPV感染在维、汉族妇女宫颈鳞癌之间无差异。     

Looking beyond human papillomavirus (HPV) genotype 16 and 18: Defining HPV genotype distribution in cervical cancers in Australia prior to vaccination

Australia has implemented a high‐coverage HPV vaccination program but has not, to date, established the distribution of HPV types that occur in cervical cancers in Australia. This information is important for determining the potential for cervical cancer prevention with both current and broader spectrum HPV vaccines. We analysed 847 cervical cancers diagnosed 2005 to 2015 in tertiary centres in the three most populous Australian states with resolution of specimens containing multiple HPV types using laser‐capture microdissection. Archived FFPE tissue was reviewed by specialist pathologists, sandwich sectioned, and initially whole‐tissue sections genotyped for HPV. Samples were first genotyped using SPF10‐LiPA25 (version 1). Negative samples were screened with DNA ELISA kit HPV SPF10, followed by genotyping with SPF+ LiPA if ELISA positive. If still negative, samples were tested on a qPCR assay targeting the E6 region of HPV16, 18, 45 and 33. Of the 847 cancers (65.1% squamous, 28.7% adenocarcinoma, 4.3% adenosquamous, 2.0% other), 92.9% had HPV detected. Of the HPV‐positive cancers, 607 of 787 (77.1%) contained HPV16 or 18, 125 of 787 (15.9%) contained HPV31/33/45/52 or 58, and 55 (7.0%) another HPV type. There was a strong correlation between HPV type and age, with younger women most likely to have HPV16/18 detected and least likely HPV negative. Our findings indicate that cervical cancers diagnosed in Australia more frequently contain HPV16/18 than in international series. This could be due to cervical screening in Australia increasing the proportion of adenocarcinomas, in which types 18 and 16 more strongly predominate, due to prevention of squamous cancers.     

Incidence risk of cervical intraepithelial neoplasia 3 or more severe lesions is a function of human papillomavirus genotypes and severity of cytological and histological abnormalities in adult Japanese women

We examined incidence probabilities of cervical intraepithelial neoplasia 3 (CIN3) or more severe lesions (CIN3+) in 1,467 adult Japanese women with abnormal cytology in relation to seven common human papillomavirus (HPV) infections (16/18/31/33/35/52/58) between April 2000 and March 2008. Sixty‐seven patients with multiple HPV infection were excluded from the risk factor analysis. Incidence of CIN3+ in 1,400 patients including 68 with ASCUS, 969 with low grade squamous intraepithelial lesion (LSIL), 132 with HSIL without histology‐proven CIN2 (HSIL/CIN2(?)) and 231 with HSIL with histology‐proven CIN2 (HSIL/CIN2(+)) was investigated. In both high grade squamous intraepithelial lesion (HSIL)/CIN2(?) and HSIL/CIN2(+), HPV16/18/33 was associated with a significantly earlier and higher incidence of CIN3+ than HPV31/35/52/58 (p = 0.049 and p = 0.0060, respectively). This association was also observed in LSIL (p = 0.0002). The 1‐year cumulative incidence rate (CIR) of CIN3+ in HSIL/CIN2(?) and HSIL/CIN2(+) according to HPV genotypes (16/18/33 vs. 31/35/52/58) were 27.1% vs. 7.5% and 46.6% vs. 19.2%, respectively. In contrast, progression of HSIL/CIN2(+) to CIN3+ was infrequent when HPV DNA was undetected: 0% of 1‐year CIR and 8.1% of 5‐year CIR. All cervical cancer occurred in HSIL cases of seven high‐risk HPVs (11/198) but not in cases of other HPV or undetectable/negative‐HPV (0/165) (p = 0.0013). In conclusion, incidence of CIN3+ depends on HPV genotypes, severity of cytological abnormalities and histology of CIN2. HSIL/CIN2(+) associated with HPV16/18/33 may justify early therapeutic intervention, while HSIL/CIN2(?) harboring these HPV genotypes needs close observation to detect incidence of CIN3+. A therapeutic intervention is not indicated for CIN2 without HPV DNA.