运用导流杂交基因芯片技术在重庆地区人乳头瘤病毒感染分型检测的研究

目的 探讨运用导流杂交基因芯片技术(HybriMax)在临床人乳头瘤病毒(HPC)感染分型检测中的运用,调查重庆地区宫颈病变患者HPV病毒分型情况进行.方法 采用导流杂交基因芯片技术,对473例宫颈病变患者以病理学诊断为金标准对宫颈病变进行分组后,进行生殖道HPV感染分型检测.结果 17种HPV亚型被检出,总检出率为63.0%(284/473),检出率较高的亚型为HPV16(23.7%、112/473)、HPV58(12.7%、60/473)、HPV53(7.4%、35/473).HPV感染率随宫颈病变SPI、CIN I、CIN II、CIN III的变化而升高(X2=77.06、P＜0.01).结论 重庆地区最常见的HPV型别是HPV16、58型.导流杂交基因芯片技术能够时检测HR-HPV和LR-HPV,可判断多型感染;用于临床诊断和普查均为理想方法.     

人乳头状瘤病毒E6/E7mRNA检测在宫颈癌早期筛查中的价值

目的 探讨人乳头状瘤病毒（HPV）E6/E7 mRNA检测在宫颈癌早期筛查中的价值.方法对700例疑似患者进行HPV E6/E7 mRNA及导流杂交基因芯片分型检测,对其阳性检出率、敏感性、特异性进行分析评价.结果 HPV导流杂交基因芯片分型阳性检出率80.0%,HPV E6/E7 mRNA阳性检出率37.3%（P〈0.05）;在3个宫颈细胞病理学分级中,HPV导流杂交基因芯片分型与HPV E6/E7 mRNA检测的阳性率随级别增加而增高,但基因芯片分型检测更为明显（P〈0.05）;随访分析中HPV E6/E7 mRNA检测的特异性、阳性预测值均明显优于HPV导流杂交基因芯片分型.结论在宫颈癌筛查中,HPV导流杂交基因芯片分型结合HPV E6/E7 mRNA检测对疾病进一步明确诊断具有较大意义.     

某区PCR法检测HPV感染的临床意义及分析

目的探讨闽北建阳区PCR-反向点杂交法在检查患者HPV感染病变中的临床意义及分析。方法回顾性分析我院检测的1624例HPV感染患者的临床资料,采用PCR-反向点杂交法对病变标本进行HPV分型检测。结果共进行1624次HPV分型检测,检出感染患者504例,检出率为31.0%,感染较多的是16、52、58、81型,并以高危型感染为主。单种分型感染共346例,两种及以上感染158例。结论 PCR-反向点杂交法在HPV感染的诊断率较高,特异性较强,对闽北地区HPV感染病变的早期发现,判断预后及指导治疗有重要价值。     

贵州省部分少数民族妇女宫颈癌的HPV谱研究

目的 探讨人乳头瘤病毒(human papilloma virus,HPV)各亚型在贵州省部分地区少数民族宫颈癌患者中的型别分布情况.方法 采用导流杂交基因芯片技术,对145例贵州省少数民族宫颈癌患者石蜡组织标本中的HPV 21种亚型(包括13种高危亚型、5种低危亚型和3种中国人群常见亚型)进行检测,并对HPV感染率及...     

某地区人乳头瘤病毒感染基因亚型的分布特点

目的了解渝北区妇女HPV感染率、基因亚型和年龄的分布情况。方法采用导流杂交法检测1181例女性宫颈脱落细胞,对其进行HPV基因亚型检测。结果 1181例标本中,感染数215例,感染率18.20%,高危亚型居前五位的依次为52、16、39、18和58型;高危型HPV感染率高于低危型,单一感染率高于多重型感染(P<0.05);各年龄组感染无显著性差异(P>0.05)。结论 HPV感染具有年轻化趋势,HPV基因分型检测对疫苗的制备与宫颈癌的防治有指导意义。     

医师取样与受检者自取样HPVDNA检测对比分析

目的采取医师取样及自取样对住院初治宫颈癌病人进行HPV DNA分型检测筛查,比较两种方法HPV检测符合率。方法随机选择宫颈癌初治患者15例,采用自身对照,使用导流杂交基因芯片技术检验技术,比较医师取样及白取样进行HPV DNA分型检测的结果。结果医师取样及受检者自取样进行HPV DNA分型检测的符合率为93．75％。结论受检者自取样如果方法得当,具有与医师取样几乎相同的检出率。     

萍乡地区人乳头瘤病毒感染现状调查

目的了解萍乡地区妇女HPV感染率和亚型分布特点及与HPV感染相关的因素。方法从2009年8月-2012年月12月．采用本院体检和门诊共1336例．进行了13种HPV高危型检测,发现阳性者进行了导流杂交基因芯片分型检测。结果1336例中,发现HPV感染者326例,总感染率为2414％。高危阳性者182例采用基因分析．全部为阳性,共检出13种HPV亚型,其中检出单一感染42例,占23．1％,混合型感染140例,占76．9％。高危型中HPV16是最常见,其次是HPV33、HPV58、HPV52和HPV68型各占2到5位。不同年龄妇女HPV感染也存在一定的差异,36—40岁、41—45岁、46—50岁检出率高于其他年龄段分别为6．1％、5．1％和3．4％。在11例浸润癌中,发现感染HPV16的有6例．HPV 33有3例。HPV 31有1例：在11例CINⅢ中HPV 16有9例,感染HPV33有2例。结论萍乡地区妇女宫颈HPV感染最常见的高危型是HPV16、HPV33、HPV58、HPV52和HPV68。     

基因芯片检测宫颈中人乳头状病毒的感染

目的采用自制HPV基因芯片技术HPV病毒各亚型在已婚妇女宫颈中的感染状况。方法选择门诊常规筛查女性638人,收集宫颈脱落细胞标本,同时进行TCT检测及自制HPV基因芯片分型检测。结果 HPV感染率43.0%,常见亚型低危型HPV6,高危型HPV16、52、33,HPV感染在各年龄组的感染率无显著差异。高危型HPV阳性率随TCT病变的严重程度而增加,差异有统计学意义（P〈0.05）。结论 HPV基因芯片结合TCT做宫颈病变筛查经济快速,对已婚各年龄组女性的HPV感染都应重视。     

石家庄市女性HPV感染情况分析

目的研究石家庄市女性宫颈脱落细胞中人乳头瘤病毒（HPV）的感染状况及其特点。方法采用核酸分子快速导流杂交技术对石家庄市2234例女性患者的宫颈脱落细胞进行HPV基因型检测。结果所测标本中HPV阳性检出率为36．3％（812／2234）,21种基因型均被检出,其中高危型651例,低危型161例。HPV高危型中,阳性率最高的是HPV16型,其他主要基因型依次是HPV52、58、53、18、66、68、31、33、39型;低危型主要为6、11、81（CP8304）型;单一HPV基因型感染的阳性率为17．1％（381／2234）,多重HPV基因型感染的阳性率为12．1％（270／2234）。结论石家庄市女性宫颈脱落细胞HPV感染率为36．3％,HPV16、52、58型是主要的感染基因型。     

高危型HPV-DNA检测在宫颈病变筛查中的应用价值

目的:探讨高危型人乳头瘤病毒(HR-HPV)检测在宫颈病变筛查中的应用价值。方法:应用第二代杂交捕获法(HC-Ⅱ)和液基薄层细胞学(TCT)对839例受检者同时行13种高危型HPV DNA检测和宫颈细胞学检查;HPV阳性和细胞学结果为非典型鳞状细胞(ASCUS)及以上患者行阴道镜下病理活检,分析HPV感染与宫颈上皮内病变的关系。结果:HPV检出率随宫颈病变程度加重呈上升趋势,ASCUS中HPV检测阳性率为75.47%,HPV阳性的ASCUS患者中病理活检47.17%为宫颈上皮内瘤变或更严重病变。结论:HPV检测在宫颈病变筛查中可作为细胞学的辅助手段提高诊断准确性,并可指导HPV感染者的治疗及用于患者的追踪随访。     

A method for removing contaminating protein during purification of human papillomavirus type 18 L1 protein from <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Human papillomavirus (HPV) types 16 and 18 are the main targets in the field of prophylactic vaccines for preventing cervical cancer. L1 protein, the major capsid protein of HPV, selfassembles into virus-like particles (VLP), which are the major component of prophylactic vaccines. To obtain highly purified L1 protein, contaminants must be removed by several chromatography steps. However, this requires a great deal of time and labor, and results in loss of large amounts of the target protein. Therefore, we have sought to develop an efficient method for removing contaminants prior to chromatography during the purification of HPV18 L1 protein from Saccharomyces cerevisiae. For this purpose the contaminating proteins were removed by an ammonium sulfate precipitation step and further removed by a removal of precipitated contaminants step. Purification of the L1 protein by chromatography was significantly improved by the removal of precipitated contaminants step. In the present work we developed two one-step chromatography methods (heparin and cation-exchange chromatography), and HPV18 L1 proteins purified by both methods self-assembled into VLP. The two chromatographic purification methods are simpler and more convenient than previous methods and are widely applicable to work with VLPs.     

尖锐湿疣患者HPV基因型检测与预后复发相关性研究

目的了解尖锐湿疣患者HPV基因型的分布情况，并观察免疫治疗后患者预后复发情况与HPV型别是否具有相关性。方法130例尖锐湿疣患者均行电灼术清除疣体，并采用导流杂交基因芯片分型技术对皮损处的疣体进行HPV的基因分型测定，术后予卡介菌多糖核酸肌肉注射2个疗程，治疗后半年内记录复发情况及复发患者的HPV基因类型。结果130例尖锐湿疣患者HPV—DNA阳性率达94．62％；共检测出13种HPV基因类型，其中低危型HPV6（47．69％）、HPV11（27．69％）所占比例较高，多重感染率达24．62％。免疫治疗后，HPV多重感染患者复发率（43．75％）明显高于单一型感染患者（26．31％），两者间差异有显著性（P〈0．05）.结论尖锐湿疣患者以HPV6、11型感染为主，HPV基因多重感染率高，且与尖锐湿疣的复发有一定相关性。     

Quadrivalent human papillomavirus (types 6, 11, 16, 18) recombinant vaccine (Gardasil®): a review of its use in the prevention of premalignant genital lesions, genital cancer and genital warts in women

Quadrivalent human papilloma virus (HPV) [types 6, 11, 16, 18] recombinant vaccine (Gardasil?; Silgard?) is composed of virus-like particles (VLPs) formed by self-assembly of recombinant L1 capsid protein from each of HPV types 6, 11, 16 and 18. The VLPs are noninfectious, containing no DNA, and are highly immunogenic, inducing high levels of neutralizing antibodies against the particular HPV types when administered to animals or humans. Quadrivalent HPV vaccine is indicated for use from the age of 9 years for the prevention of premalignant genital lesions (cervical, vulvar and vaginal), cervical cancer and external genital warts (condyloma acuminata) causally related to certain oncogenic or specific HPV types. In placebo-controlled clinical trials, quadrivalent HPV vaccine administered as three doses over 6 months provided high-level protection against infection or disease caused by the vaccine HPV types over 2-4 years of follow-up in females aged 15-45 years who were naive to the vaccine HPV types. A degree of cross-protection against certain other non-vaccine high-risk HPV types was also observed. The vaccine is not effective against current infection with a vaccine HPV type. Girls or women with current infection with one or more of the vaccine HPV types gained protection from infection or disease caused by the remaining vaccine HPV types and they were also protected against reinfection with the same HPV type after clearance of an infection caused by a vaccine HPV type. High seroconversion rates and high levels of anti-HPV antibodies were observed in all vaccinated individuals of all age ranges from 9 to 45 years. No correlation was found between antibody levels and protective efficacy of the vaccine. Rechallenge with quadrivalent HPV vaccine produced a potent anamnestic humoral immune response. The vaccine is generally well tolerated and is projected to be cost effective in most pharmacoeconomic models. Therefore, quadrivalent HPV vaccine offers an effective means, in combination with screening programmes, to substantially reduce the burden of HPV-related precancerous lesions and cancer, particularly cervical cancer, as well as anogenital warts.     

High-risk hpv infection in sexually active and non active young women

To determine the prevalence of high-risk HPV infection before first vaginal intercourse and after initiation of sexual activity in 14-22 years old girls, to find which HPV types occur most commonly. During the gynecologic exam cervical swabs were collected. Specimens were tested using Hybrid Capture 2 DNA test. In positive samples digene HPV genotyping test was performed. Subjects included 264 women, of whom 169 (64.0%) reported previous sexual activity and 95 (36.0%) had not yet had vaginal intercourse. There were 43 cases (16.3%) of high risk HPV, with 39 cases in those reporting sexual activity and 4 in virgins. Single strain HPV infection was detected in 62.8%, two strains in 23.2%, and three strains in 14.0% of subjects. HPV-16 was the most common type (20.9%), next more prevalent types were 18, 56, 31, 33 and 59. HPV-16 or HPV-18 were detected in 39.5% of the subjects. Not a single case was identified containing both types. The prevalence of high-risk HPV is common in sexually active young Lithuanian women. HPV were found in a small percentage of girls who had never had vaginal intercourse. The prevalence of types 16 and 18 among Lithuanian women was higher than in international data.     

深圳市龙华地区人乳头瘤病毒感染率及亚型分析

目的了解深圳市龙华区某医院及其所覆盖的社康中心就诊女性人乳头瘤病毒(HPV)感染率、年龄及基因亚型分布的最新数据,以期为建立和完善当地女性HPV监测、HPV疫苗开发和宫颈癌防治系统提供科学依据。方法选取最近1年内在我院总部及其所覆盖的29个社康中心接受宫颈HPV检查的12658例女性为研究对象,按照年龄分为10组(每5岁为一组),采用PCR-膜杂交法检测28种HPV基因亚型。结果12658例检测样本中,HPV阳性2202例,总感染率17.40%。HPV亚型感染率排名前五位的分别是:HPV52、HPV16、HPV58、HPV51和HPV53。低危型、疑高危型、高危型感染率分别为6.56%(830/12658)、1.88%(238/12658)和15.67%(1984/12658)。16~20岁年龄组女性HPV感染率(48.44%)及HPV高危型感染率(58.59%)均最高,46~50岁年龄组女性HPV感染率(12.26%)及HPV高危型感染率(9.75%)均最低。多重感染方面,以单一基因感染为主,感染率为12.68%(1605/12658),两种基因型感染率为3.38%(428/12658),三种及以上基因型感染率为1.34%(169/12658),差异具有统计学意义(P<0.05)。结论该地区就诊女性HPV感染以高危型为主,年轻组(16~25岁)和中老年组(≥56岁)为高发人群,临床应采用针对性措施,推广HPV多价疫苗的应用,同时加强对年轻女性和中老年女性的卫生宣传、保健教育等工作以及对病毒感染的监测力度。     

衡阳地区HIV阳性妇女宫颈HPV感染情况研究

目的了解本地区人类免疫缺陷病毒（HIV）阳性女性宫颈人乳头瘤病毒（HPV）感染状况,为HIV阳性人群中HPV感染的防治提供依据。方法采用ELISA法和胶体金法检测4576例女性患者血清或血浆中的HIV抗体,两种方法均为阳性标本再送检确认。PCR＋膜杂交法检测病人宫颈脱落细胞及宫颈粘液标本进行HPV病毒基因分型。结果 4576例病例中,HIV感染率为0.83%;HPV感染率为21.02%。HIV阳性组中HPV感染率为60.53%,HIV阴性组中HPV感染率20.69%,差异有统计学意义（χ^2=36.02,P〈0.005）。在962例HPV阳性病例中,HIV阳性组的混合HPV感染率为56.52%;HIV阴性组的混合HPV感染率为20.98%;差异有统计学意义（χ^2=16.62,P〈0.005）。结论 HIV阳性妇女的宫颈HPV感染率高,且高危与多重HPV感染常见,提示HPV感染与HIV感染患者关系密切。     

Comparison of the immunogenicity of the human papillomavirus (HPV)-16/18 vaccine and the HPV-6/11/16/18 vaccine for oncogenic non-vaccine types HPV-31 and HPV-45 in healthy women aged 18-45 years

Protection against oncogenic non-vaccine types (cross-protection) offered by human papillomavirus (HPV) vaccines may provide a significant medical benefit. Available clinical efficacy data suggest the two licensed vaccines (HPV-16/18 vaccine, GlaxoSmithKline Biologicals (GSK), and HPV-6/11/16/18 vaccine, Merck & Co., Inc.) differ in terms of protection against oncogenic non-vaccine HPV types -31/45. The immune responses induced by the two vaccines against these two non-vaccine HPV types (cross-reactivity) was compared in an observer-blind study up to Month 24 (18 mo post-vaccination), in women HPV DNA-negative and seronegative prior to vaccination for the HPV type analyzed (HPV-010 [NCT00423046]). Geometric mean antibody titers (GMTs) measured by pseudovirion-based neutralization assay (PBNA) and enzyme-linked immunosorbent assay (ELISA) were similar between vaccines for HPV-31/45. Seropositivity rates for HPV-31 were also similar between vaccines; however, there was a trend for higher seropositivity with the HPV-16/18 vaccine (13.0-16.7%) versus the HPV-6/11/16/18 vaccine (0.0-5.0%) for HPV-45 with PBNA, but not ELISA. HPV-31/45 cross-reactive memory B-cell responses were comparable between vaccines. Circulating antigen-specific CD4+ T-cell frequencies were higher for the HPV-16/18 vaccine than the HPV-6/11/16/18 vaccine (HPV-31 [geometric mean ratio [GMR] =2.0; p=0.0002] and HPV-45 [GMR=2.6; p=0.0092]), as were the proportion of T-cell responders (HPV-31, p=0.0009; HPV-45, p=0.0793). In conclusion, immune response to oncogenic non-vaccine HPV types -31/45 was generally similar for both vaccines with the exception of T-cell response which was higher with the HPV-16/18 vaccine. Considering the differences in cross-protective efficacy between the two vaccines, the results might provide insights into the underlying mechanism(s) of protection.     

常熟地区女性HPV感染及亚型分布调查研究

目的 调查江苏常熟地区女性人乳头瘤病毒（HPV）感染及其亚型的分布情况.方法应用DNA导流杂交技术对1 602例妇科门诊患者进行HPV基因分型检测.结果 1 602例患者中阳性患者为719例,感染率为44.9%,共检出20种HPV型别,其中单一感染者498例,占阳性人数的69.3%,二型感染者162例,占阳性人数的22.5%,三型或三型以上者59例,占阳性人数的8.2%,阳性患者中高危型的依次以16,52,68,31,33为主,低危型以11、6为主.结论 常熟地区妇女感染HPV率较高,以单一感染为主,主要为16型,其次为52,68,31,33型.作为宫颈癌筛查的有效手段,一次PCR检测多种HPV亚型的分子杂交技术有利于对HPV亚型多重感染的诊断和宫颈癌的防治有重要意义.     

皮肤性病 60例的人乳头瘤病毒感染型别分析

目的 探讨皮肤性病病人不同病变类型中的人乳头瘤病毒(HPV)感染型别的检测和分析结果.方法 选取淮南市第一人民医院2018年2月至2019年2月收治的60例皮肤性病病人作为研究对象,对所有病人均利用PCR-反向斑点杂交法进行HPV-DNA分型的检测,并对皮肤性病病人不同病变类型中HPV感染的型别进行统计和比较.结果 本研究60例标本中HPV-DNA检测阳性例数为71.67％(43/60);其中HPV单一型的31例,占比72.09％(31/43);HPV2种或以上者共12例,占比27.91％(12/43).43例HPV检测阳性标本中高危型检出例数最高,为24例,占比55.81％;低危型13例,占比30.23％;混合型6例,占比13.95％.本研究不同病变类型皮肤性病中,以尖锐湿疣的HPV感染阳性率最高,为76.47％(26/30);鲍温样丘疹病次之阳性率为66.67％(10/15);其次为冠状沟珍珠疹和假性湿疣,阳性率分别为50％(4/8)和42.86％(3/7).在HPV感染型别上,以尖锐湿疣类别最多,包括了6、11、16、18、31、33、35、43和66,而鲍温样丘疹病主要为16,男性冠状沟珍珠疹主要为低危型的6、42、43和81,女性假性湿疣也为低危型的6、42、83.结论 皮肤性病病人HPV感染存在不同的类别,因此临床上在诊断时需要积极开展相应的检测工作,从而为临床治疗方案的设定和预后结果的判断提供参考依据.     

Identification of human papillomavirus DNA in melanoma biopsy specimens of Greek population

The aim of this retrospective study was to evaluate melanoma biopsy specimens from the Greek population living in the prefecture of Larissa for the presence of human papillomavirus (HPV) DNA and to determine the possible relationship between HPV and clinical outcome in these patients. Twenty-eight melanoma biopsy specimens, 20 from primary cutaneous melanoma and eight from melanoma metastasis were obtained from 28 patients. The biopsy samples were formalin-fixed and paraffin wax-embedded. The control group consisted of three junctional melanocytic nevi, histologically confirmed, and three punch biopsies from normal skin that were obtained from six healthy individuals. The presence and types of HPV DNA were assessed by the amplification of a fragment of the LI region by consensus primer polymerase chain reaction (PCR) combined with restriction fragment length polymorphism analysis (RFLPA). In each biopsy specimen that was evaluated, HPV 6, HPV 11, HPV 16 and HPV 18 positive controls from genital HPV lesions were included. Five of 28 (17.85%) biopsy melanoma specimens were positive for HPV DNA. Conversely, HPV was not detected in any of the biopsy specimens of the control group (0/6). HPV viral type 16 was found in two samples and HPV 6 DNA in three. Our results regarding the possible relationship between melanoma and HPV DNA were not statistically significant (p > 0.05). These findings suggest that ultraviolet sun exposure remains the main cause of melanoma in our region. The role of cutaneous HPV infection in the pathogenesis of melanoma remains elusive.