第二代杂交捕获法检测宫颈人乳头瘤病毒感染的初步研究

目的:探讨人乳头瘤病毒与宫颈癌及癌前期病变的相关性.方法:选取2004年12月～2005年7月我院妇科宫颈病门诊妇女1689例,取宫颈刷出物同时作宫颈薄层液基细胞学涂片与HPV-DNA检测,HPV-DNA检测方法是第二代杂交捕获法,可一次检测13种高危型HPV病毒,分析HPV感染与宫颈癌及癌前期病变的关系.结果:HPV-DNA检出率随宫颈癌变程度加重呈趋势性升高,1689例病人中高危型HPV检出率为25.16%(425/1689),宫颈癌及癌前期病变中为90.35%(234/259).结论:高危型HPV是诱发宫颈癌及癌前期病变的重要病因学因素.第二代杂交捕获法可作为宫颈癌的一种筛查方法.     

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.     

HPV testing in the evaluation of the minimally abnormal Papanicolaou smear.

Minor cytologic abnormalities of the cervix, such as atypical squamous cells of undetermined significance (ASCUS), are vastly more common than high-grade squamous intraepithelial lesions or invasive cancer. Current guidelines for the management of ASCUS include repeating the Papanicolaou (Pap) smear at specific intervals, referring all patients for colposcopy or using an adjunctive test such as hybrid capture human papillomavirus (HPV) testing or cervicography. The usefulness of the Pap smear is limited by its considerable false-negative rate and its dependence on clinician and laboratory performance. Colposcopy is a highly sensitive procedure, but many patients with ASCUS have normal colposcopic findings. The hybrid capture test not only measures quantitative HPV load but also detects both oncogenic and nononcogenic HPV types, thereby increasing the probability that serious cervical disease is not missed. Hybrid capture sampling is simple to perform, and positive results are strongly associated with cervical dysplasia. HPV testing in women with ASCUS can be used as an adjunctive test to identify those with HPV-associated disease; it can also serve as a quality assurance measure. Together, repeat Pap smears and HPV testing should identify most patients with underlying cervical dysplasia. Combined testing may also minimize the number of unnecessary colposcopic examinations in women who have no disease.     

Multiple biomarkers in molecular oncology. I. Molecular diagnostics applications in cervical cancer detection

The screening for cervical carcinoma and its malignant precursors (cervical neoplasia) currently employs morphology-based detection methods (Papanicolaou [Pap] smear) in addition to the detection of high-risk human papillomavirus. The combination of the Pap smear with human papillomavirus testing has achieved significant improvements in sensitivity for the detection of cervical disease. Diagnosis of cervical neoplasia is dependent upon histology assessment of cervical biopsy specimens. Attempts to improve the specificity of cervical disease screening have focused on the investigation of molecular biomarkers for adjunctive use in combination with the Pap smear. Active research into the genomic and proteomic alterations that occur during human papillomavirus-induced neoplastic transformation have begun to characterize some of the basic mechanisms inherent to the disease process of cervical cancer development. This research continues to demonstrate the complexity of multiple genomic and proteomic alterations that accumulate during the tumorigenesis process. Despite this diversity, basic patterns of uncontrolled signal transduction, cell cycle deregulation, activation of DNA replication and altered extracellular matrix interactions are beginning to emerge as common features inherent to cervical cancer development. Some of these gene or protein expression alterations have been investigated as potential biomarkers for screening and diagnostics applications. The contribution of multiple gene alterations in the development of cervical cancer suggests that the application of multiple biomarker panels has the potential to develop clinically useful molecular diagnostics. In this review, the application of biomarkers for the improvement of sensitivity and specificity of the detection of cervical neoplasia within cytology specimens will be discussed.     

贵州地区宫颈癌机会性筛查资料分析

目的探讨贵州地区进行宫颈癌机会性筛查的价值。方法回顾性分析2010年11月-2011年10月贵州省人民医院妇科门诊及住院进行宫颈癌机会性筛查的1 842例患者资料,筛查方法包括液基细胞学技术、宫颈刮片、人乳头瘤病毒(HPV)分型检测、HPV第2代捕获杂交法检测、阴道镜检查,以病理确诊宫颈癌前病变及宫颈癌。结果贵州地区女性人群的HPV平均感染率为21.87%。共检出宫颈癌前病变39例(2.12%),宫颈癌2例(0.11%)。结论医院内就诊女性宫颈病变检出率高,应高度重视其机会性筛查,有助于早期干预治疗。     

HPV分型检测和TCT检查在宫颈病变筛查中的应用

目的：探讨人乳头瘤病毒（HPV）分型检测和液基细胞学（TCT）检查在筛查宫颈病变中的应用。方法：回顾性分析2012.9~2013.9在我院妇科门诊就诊,同时采用HPV分型和TCT检查的门诊妇女宫颈脱落细胞标本1128例,并最终进行组织病理学检查。结果：1128例标本中HPV阳性率为64.4%（726/1128）,共检出21种亚型,高危亚型16种,低危亚型5种,高危型以16,58,52多见,构成比分别为25.0%,12.1%和10.2%。低危型以61,11型多见,构成比分别为3.4%和1.9%。其中单一感染555例,多重感染171例,最多为四重感染。TCT检查≥ASCUS为87.8%（990/1128）,在ASCUS、LSIL、HSIL和SCC中HPV的检出率分别为60.3%、71.8%、80.3%和100%。HPV分型检测（H组）的阳性预测值为74.1%,阴性预测值为68.7%,TCT检查（T组）的阳性预测值和阴性预测值分别为63.2%（626/990）和72.5%（100/138）,两者联合检测（H＋T组）阳性预测值和阴性预测值分别为75.5%（100/138）、100%（100/100）。H组和H＋T组的阳性预测值高于T组（X2值分别为22.690,28.822,P值均为0.000）,H＋T组的阴性预测值高于H组和T组（X2值41.847,32.768,P值均为0.000）。结论： HPV分型检测是准确性高并能明确基因类型及多重感染的检测,联合TCT检查更能有效的筛查宫颈病变细胞,为临床防治宫颈癌及青岛地区HPV疫苗使用提供更可靠地科学依据。     

HPV感染流行病学及其与生殖系统其他疾病相关性研究

目的了解本地区女性生殖道人乳头瘤病毒(HPV)感染分布特征。方法收集2 509例16~79岁女性就诊者及体检者宫颈脱落细胞,采用核酸分子杂交基因分型技术进行HPV分型检测,同时进行液基薄层细胞学检查、白带常规检查等其他检查,分析HPV整体及主要亚型检测结果,以及HPV感染和其他生殖系统疾病的关系。结果 HPV总检出率为13.9%,高危型HPV检出率为10.4%,低危型HPV检出率为3.3%。常见HPV高危型为16、58、18、33和56亚型;常见低危型为11、6和43亚型。感染高峰年龄段为16~20岁、55~60岁,以单一感染为主。宫颈炎患者、阴道炎合并宫颈炎患者HPV检出率高于健康者(P0.05);解脲脲原体阳性患者HPV检出率高于阴性患者(P0.05)。结论 HPV检出率,尤其是16、58和33亚型检出率较高,和其他生殖道感染性疾病有一定的相关性。     

Clinical experience with the Cervista HPV HR assay: correlation of cytology and HPV status from 56,501 specimens

Testing for high-risk (HR) human papillomavirus (HPV) is a key component of current recommendations for cervical cancer screening. Herein is described our clinical experience using Cervista HPV HR, a testing platform recently approved by the US Food and Drug Administration for clinical use. Using data from a high-volume commercial laboratory, a retrospective analysis of cytologic and Cervista HPV HR test results from 56,501 samples was performed, and an indirect comparison was made with previous experience with 53,008 samples tested using the Hybrid Capture 2 platform. Of samples analyzed using Cervista HPV HR, 1.5% were of insufficient volume for testing and 1.1% yielded an insufficient signal from the internal control to be reported. In samples with a cytological interpretation of atypical squamous cells of undetermined significance, 48.5% (95% confidence interval [CI], 47.5 to 49.5) tested positive using Cervista HPV HR, compared with 59.4% (95% CI, 58.3 to 60.5) of samples using Hybrid Capture 2. Of samples from women aged 30 years or older with a negative cytological interpretation, 5.8% (95% CI, 5.6 to 6.1) tested positive using Cervista HPV HR, compared with 5.5% (95% CI, 5.3 to 5.7) of samples using Hybrid Capture 2. When stratified by five-year age groups between 30 and 65 years, positivity rates for high-risk human papillomavirus were similar in the Cervista HPV HR and Hybrid Capture 2 populations, and were consistent with expectations established by the literature.     

1268例慢性宫颈炎患者人乳头瘤病毒感染与基因型分布

目的调查慢性宫颈炎患者人乳头瘤病毒(HPV)感染及其基因型的流行分布状况。方法采用基因芯片技术对1268例慢性宫颈炎患者宫颈分泌物进行18种高危型HPV基因型和5种低危型HPV基因型的检测,分析其流行分布特点。结果 HPV阳性430例,占33.91%,高危型HPV感染占HPV阳性的82.93%,以HPV16为主,其次为HPV58、HPV18、HPV56。低危型感染占17.07%,以HPV11最高,其次为HPV6;单一感染占74.65%,重复感染占25.35%;高危型复合感染占66.06%,低危型+高危型复合感染占31.19%,单纯低危型的复合感染占2.75%。慢性宫颈炎患者HPV感染率以50岁组最高。结论慢性宫颈炎患者HPV的感染率较高,以高危型HPV基因型为主,临床应重视HPV感染的筛查、监测与治疗,防治宫颈癌。     

Human papillomavirus, cervical cancer, and the vaccines

How are human papillomavirus (HPV), cervical cancer, and the recently developed HPV vaccines associated with each other? Human papillomavirus is a highly prevalent infection that is easily and unknowingly transmitted because of its asymptomatic nature and long incubation period. Infection requires skin-to-skin contact and is typically sexually transmitted. More than one-half of sexually active women acquire HPV, making it the most prevalent sexually transmitted disease. Cervical cancer ranks second in deaths from cancer among women in developing countries and kills nearly 4000 women in the United States annually. Several types of HPV have been strongly linked to causing cervical cancer and genital warts. Those causing cervical cancer are considered high-risk types and those causing genital warts are considered low-risk types. Until recently, prevention strategies included abstinence, condom usage, and early detection with a Papanicolaou test (Pap smear). New developments have led to 2 vaccines aimed at preventing the viral infection. One is a quadrivalent vaccine preventing infection from 4 HPV types (HPV types 6, 11, 16, and 18) (Gardasil). It is approved in the United States and Europe for the prevention of HPV-associated cervical cancers and genital warts in females between the ages of 9 and 26 years old. The second is a bivalent vaccine preventing infection from 2 high-risk oncogenic HPV types (HPV types 16 and 18) (Cervarix). It is currently under study and not yet available in the United States. Both vaccines have proven highly effective at preventing infection from their corresponding HPV types. Of importance, neither vaccine is to be used for treatment. Vaccination does not replace routine cervical cancer screening with Pap smears, as the vaccines do not protect against all HPV types.     

PS1-25: Cervical Cancer Testing and Follow-up in Four Managed Care Plans, 1998-2007

Background/Aims Cervical cancer screening is performed to detect pre-cancerous cervical intraepithelial neoplasia or invasive cancerous cervical lesions prior to the onset of symptoms so they can be removed before the cancer has developed or spread. With the addition of high-risk human papillomavirus (HPV) testing to the long-established Papanicolaou (Pap) smear, national and health plan screening guidelines have been regularly updated in recent years. Guidelines on screening frequency and follow-up protocols have also changed over time and will likely continue to do so as HPV vaccination becomes more widespread. Here we describe patterns and results of cervical cancer testing and follow-up over a 10-year period within four geographically-dispersed U.S. managed care organizations. Methods Using data collected by the SEARCH: Screening Effectiveness And Research in Community-Based Healthcare project, we analyzed electronic medical record data on all women aged 20-65 during the period 1998-2007 across four HMORN sites. We created standardized files for Pap smear dates and results; cervical histology dates, types, and results; and HPV test dates and results. We also collected Virtual Data Warehouse data on HPV vaccinations, and selected diagnosis and procedure codes. We calculated rates of Pap testing, HPV testing, colposcopy, and cervical histology (biopsy and treatment). We also calculated rates for Pap testing we classified as "screening." Among women who had Pap tests in 2002 and 2007 and no abnormal test directly preceding the index test, we examined patterns of screening frequency. We also examined frequencies and trends in the results of "screening" Pap testing and cervical histology. Results Overall, annual Pap testing rates decreased and HPV testing rates dramatically increased over the 10 years, while rates of colposcopy, cervical histology, and cervical treatment did not display obvious patterns. Trends varied by age group and health plan. Pap screening frequency differed by health plan; overall, in 2007 a higher proportion of subjects had longer screening intervals (2 years or greater) than in 2002. Information on patterns of Pap and histology results will be presented. Discussion Evaluating trends in cervical cancer testing and follow-up may highlight opportunities to optimize cervical cancer screening delivery in community-based settings.     

Detection of human papillomavirus DNA by PCR/microfluorometry for screening of cervical cancer

BACKGROUND: Cervical cancer screening is conducted by a cytological Papanicolaou (Pap) test. For screening, it is becoming increasingly important to introduce a more objective result, based on human papillomavirus (HPV) DNA test. We describe here a practical method allowing the mass detection of HPV-DNA by PCR followed by fluorogenic DNA intercalation. METHODS: Samples used were cervical scrapes or biopsy specimens obtained from women who had undergone cytological testing for cervical cancer. Crude DNAs were extracted by a simplified proteinase K-boil method. Common and type-specific primers were newly designed for major types of high-risk HPVs. A fluorogenic DNA intercalator, SYBR Green I was directly added to the specific PCR products. The resultant fluorescence was measured by a conventional fluorometric microplate reader. RESULTS: The proposed PCR/microfluorometry (MFL) allowed a simple, rapid and economical detection of HPV-DNA without any use of labeling primers or probes. HPV-DNAs were found in 48.2% (123/255) of the cervical scrapes. The detection rate of HPV in cervical cancer biopsy specimen was 92.4% (61/66). CONCLUSIONS: PCR/MFL detection of HPV-DNA, followed by combined type-specific PCR, is expected to be an extremely useful tool in cervical cancer screening.     

Diagnosis of human papillomavirus gynecologic infections

The identification in the early 1980s of human papillomavirus (HPV) DNA in cervical carcinoma generated interest in molecular classification of the virus, and prompted studies regarding the oncogenic potential of genital HPVs. Subsequent studies confirming the presence of HPV in greater than 90% of precancerous cervical lesions and close to 100% of cervical cancers has raised concerns regarding the adequacy of Papanicolaou (Pap) smear testing for the detection of precancerous lesions/HPV infection. A variety of detection methods adjunctive to cytologic testing have been described, including detection at the macroscopic level, cerviography, colposcopy, and serologic and molecular-based HPV testing. Recently, there has been intense interest in molecular-based detection and typing of HPV-induced genital lesions. This has resulted in the development of a variety of molecular-based detection methods including Southern transfer, dot blotting, in situ hybridization, hybrid capture, and PCR-based assays. This article provides an overview of each of the molecular methods, and addresses the potential future role of molecular-based HPV testing.     

Screening for cervical cancer and initial treatment of patients with abnormal results from papanicolaou testing

New techniques for cervical cancer screening and a better understanding of the natural history of human papillomavirus (HPV) and cervical neoplasia have inspired a quest for more rational screening strategies for cervical cancer. Often, screening intervals for women older than 30 years can be expanded safely to every 3 years, and experts now agree that screening may cease after hysterectomy and in elderly women (provided certain criteria have been met). Liquid-based cytology produces more satisfactory specimens than conventional testing and offers the valuable option of treating atypical squamous cells of undetermined significance by "reflex" testing for high-risk types of HPV on the original specimen. Testing for HPV as an adjunct to cervical cytology for primary screening is now considered reasonable for many women older than 30 years.     

新疆地区妇女宫颈病变组织中HPV感染与分型研究

目的探讨人乳头瘤状病毒（human papillomavirus,HPV）型别在宫颈病变中分布情况。方法采用基因芯片技术对239例宫颈癌前病变或宫颈鳞癌患者的石蜡组织标本进行23种HPV基因型别检测。结果检测出20种基因型,HPV总感染率为90．0％（215／239）;单一感染中高危型HPV16感染率为84．2％（117／139）,低危型中HPV11感染检出1例;多重感染率为35．3％（76／215）,64例为HPV16与其他高危病毒的复合感染,5例为HPV16与低危病毒（6,11,43）的复合感染,7例为低危病毒与低危病毒的复合感染。结论高危型HPV16感染是新疆地区妇女宫颈病变中常见亚型,多重感染可能与宫颈病变进展无关。     

高危型HPV—DNA检测对宫颈细胞学诊断为ASC—H的分流作用

目的探讨高危型HPV—DNA检测对宫颈细胞学诊断为不典型鳞状上皮细胞不除外高度鳞状上皮内病变（ASC—H）的分流管理作用。方法对行新柏氏TCT膜式液基超薄细胞学检查,且诊断结果为ASC—H的112例患者行高危型HPV—DNA检测及阴道镜下活检：比较分析高危型HPV—DNA检测结果与病理组织学诊断结果结果112例ASC—H患者中,病理组织学结果为高级别鳞状上皮内病变以下的患者（包括CINI、宫颈慢性炎）检出率为34．82％（39／112）。HPV阳性时高级别鳞状上皮内病变以下的患者检出率为16．88％（13／77）,HPV阴性时的检出率为74．29％（26／35）;高级别鳞状上皮内病变以上的患者（包括CINII-Ⅲ、宫颈癌）检出率为65．18％（73／112）,HPV阳性时高级别鳞状上皮内病变以上的患者检出率为83．12％（64／77）,HPV阴性时的检出率为25．71％（9／35）。HPV阴性患者栓出高级别鳞状上皮内病变以下患者的概率高于HPV阳性者,差异有统计学意义（X^2=34．93,P〈0．05）。结论高危型HPV—DNA检测可有效地对ASC—H患者进行分流,有利于临床对ASC—H患者采取合理有效的处理方法。     

人乳头瘤病毒基因分型方法的建立和应用

目的 建立一种简便快捷、灵敏度高、经济实用的人乳头瘤病毒(HPV)基因分型的低密度基因芯片技术,并对该方法进行评价.方法 用低密度基因芯片技术对355例疑似HPV感染女患者检测HPV并分型;用杂交捕获Ⅱ代(HCⅡ)法进行评价.并用该法对珠江三角洲地区的730例标本进行HPV分型检测.结果 355例疑似样本中,低密度基因芯片技术和HCⅡ法分别检出211例(59.4%)和222例(62.5%)阳性标本,符合率达94.1%(334/355).低密度基因芯片技术共检测出15种常见高危型和5种低危型,其中16、52、58、56型检出率较高.结论 低密度基因芯片技术能具体分型和检测混合感染,HPV检测与细胞学检测结合.对宫颈癌筛查有重要意义.     

武汉市成年女性人乳头瘤病毒感染及基因分型检测

目的研究武汉市成年女性人乳头瘤病毒(HPV)感染及亚型分布情况。方法采用导流杂交基因芯片技术对2538例女性宫颈脱落细胞标本进行HPV基因分型检测。结果 HPV感染率14.9%(379/2538),高危型感染率12.6%(321/2538),高危型HPV阳性人群中各基因型百分比由高到低依次为52型(30.8%),16型(27.7%),58型(14.3%),18型(10.6%),≤25岁组HPV感染率、高危型感染率、混合感染率均高于其他3组。结论武汉市成年女性HPV感染状况及HPV基因型分布与其他地区差异无统计学意义。     

高危型人乳头瘤病毒在广西地区感染情况的调查分析

目的探讨高危型人乳头瘤病毒(HPV)在广西地区的感染情况。方法采用第2代杂交捕获技术混合检测13种高危型HPV-DNA,对26 796份女性宫颈分泌物标本的检测结果进行回顾性分析。结果广西地区高危型HPV感染率为17.94%,其中桂北地区HPV感染率最高,且各区域间比较差异有统计学意义(P0.05)。HPV感染的高峰年龄段在小于20岁、50~60岁、≥60岁,各年龄段HPV感染率比较差异有统计学意义(P0.05);HPV高病毒载量患者的比例为40.93%,且随着年龄的增长呈上升趋势。结论高危型HPV感染在广西地区存在地域和年龄差异,对高发地区、年龄段应重点监测,必要时对高病毒载量患者完善宫颈细胞学及组织学的检查。