尖锐湿疣患者皮损中HPV的检测分型及HPV6b L1变异分析

目的 探讨温州地区尖锐湿疣（CA）患者人乳头瘤病毒（HPV）感染型别及HPV6基因多态性。 方法 采用基因芯片检测73例CA患者皮损组织HPV 21种基因型的感染率,对所获得的11例患者的HPV6阳性者行HPV6b L1基因测序分析。 结果 73例CA患者标本中检出HPV阳性61例,总的阳性检出率为83.56％。在61例HPV阳性患者,其中单一型别阳性检出率为93.44％,单一型别感染中以HPV11（29.51％）型为主,其次为HPV6 （18.03％）型;混合型感染阳性检出率为6.56％,以HPV6+11型感染为主。此外,CA患者HPV 6b L1基因测序结果表明,6个位点的核苷酸存在多态性,其中810位点A/T变异存在于所有标本,可能为温州地方株突变。结论 HPV11、6型感染为CA的主要致病型别,HPV 6b L1基因存在多处突变。     

上海郊区10株人乳头状瘤病毒6型的L1基因序列和进化树分析

目的了解上海郊区人乳头状瘤病毒(HPV)6型L1基因在不同疣组织(尖锐湿疣和寻常疣)结构特点和变异规律,为临床治疗和预防提供理论依据。方法采用PGMY09/11通用引物扩增的方法进行HPV基因分型,8例尖锐湿疣和2例寻常疣HPV 6型阳性标本再进行L1基因直接测序;测序结果进行BLAST;然后;进一步用Geneious软件对测序结果与Gen Bank中发布的其他国家及国内的HPV6型病毒株L1基因序列比对,同时进行进化树构建。结果与参考株X00203相较共出现11种同义突变和3种错义突变。从BLAST结果和构建的系统进化树图可以看出,本地区HPV6型病毒株中,样本13639、14960、15987和15989是HPV6的A型;14300、16320、14304、15976、15982和15330是HPV6的B1型。结论本地区皮肤疣感染的HPV6型4株为A系,6株为B1亚系,与国内外HPV6型病毒株基因都有同源性背景。     

尖锐湿疣66例HPV检测及基因分型分析

目的了解贵阳地区尖锐湿疣(CA)皮损中人乳头瘤病毒(HPV)的感染状况及基因测序分析。方法提取66例CA临床样本的HPVDNA,采用MY09/MY11引物进行PCR扩增,筛选出阳性样本;分别用HPV6,11,16,18型特异性引物对阳性样本扩增分型;10例不同临床特征阳性样本,将其MY09/MY11PCR产物进行正反向测序分析。结果65例检测为HPVDNA阳性,HPV6(62,93.0%),HPV11(53,81.5%),HPV16(5,7.60%),HPV18(6,9.0%)单型别感染(13,20.0%)以HPV6为主;混合感染(52,80.0%)以HPV6+11(41,63.2%)为主;三型混合分别为HPV6+11+16(4,6.1%)和HPV6+11+18(5,7.6%)。测序分析:8例为HPV6型,有6个核苷酸位点发生突变,其中第7162位核苷酸C→T,导致P147L和7159位核苷酸A→T,导致Y146F发生错义突变;2例为HPV11型,1例与提交序列有100%的一致性,另1例有4个位点发生核苷酸突变,第7104位核苷酸缺失一个A,导致N133T错义突变。结论本地区CA主要以HPV6+11型的混合感染为主。HPV6,11型与原型序列比较均发现有碱基和氨基酸的改变。     

男性尖锐湿疣皮损HPV基因芯片型别分析

目的探讨本地区男性尖锐湿疣患者皮损中的HPV型别感染情况及其临床意义。方法采用基因芯片微阵列技术对200例男性尖锐湿疣皮损进行26种HPV基因型别检测。结果男性尖锐湿疣患者皮损中HPV检出率为99.00%(198/200),检出的198例阳性患者中单一型别感染42.42%(84/198),以HPV6/11为主(94.00%);多重HPV混合感染以高低危HPV混合感染为主(P<0.05),18～35岁组较>35岁组高危HPV感染率偏多(P<0.05),皮损部位间存在HPV型别差异(P<0.05)。皮损中检测出HPV7型感染4例和HPV67型感染1例。结论男性尖锐湿疣皮损以HPV6/11型感染为主,并多伴有其他HPV型别感染;年龄、皮损部位和感染重数影响HPV型别分布。     

人乳头瘤病毒11型早期蛋白E6和E7基因的克隆及序列分析

目的：从尖锐湿疣(CA)标本中克隆出人乳头瘤病毒11型(HPV11)早期蛋白E6、E7基因，并进行序列测定及编码氨基酸序列分析，为HPV感染的检测和基因工程疫苗研究奠定基础。方法：用PCR法，从CA标本中扩增出HPV11E6、E7基因，与载体pGEX-6P-1连接成重组质粒pGEX-6P-1／E6、pGEX-6P-1／E7，酶切鉴定及双脱氧法测序观察其变异状况。结果：克隆出HPV 11早期蛋白E6、E7基因，成功构建重组质粒pGEX-6P-1／E6、pGEX-6P-1／E7。本研究克隆出的HPV11 E7基因与GenBank标准株序列完全相同，E6基因有两个位点的变化。结论：本研究克隆出的HPV11 E6、E7基因与标准株基本相同，这将为进一步研究E6、E7基因的表达、免疫活性及流行病学奠定基础。     

自行制备的HPV6b和11型E7蛋白多克隆抗体检测尖锐湿疣皮损的研究

目的 采用自行制备的HPV6b和11型E7蛋白多克隆抗体检测外生殖器尖锐湿疣皮损组织中HPV6b和11型E7蛋白的表达,探讨该抗体在临床检验中的应用价值。 方法 55例经临床和病理确诊的尖锐湿疣皮损石蜡标本,用自行制备的HPV6b和11型E7蛋白多克隆抗体进行免疫组化检测,其中18例皮损冰冻标本用RT-PCR法测定HPV6b和11型E7蛋白mRNA表达,分析与免疫组化结果的符合率。 结果 55例尖锐湿疣皮损的免疫组化染色显示,HPV6b和11型E7蛋白为胞核染色,且皮损全层表皮细胞均有阳性表达,但基底层阳性细胞较多。HPV6b和11型E7蛋白阳性表达率分别为76.36%（42/55）和58.18%（32/55）,总阳性表达率为94.55%（52/55）,两蛋白双阳性率为40.00%（22/55）,两蛋白均阴性表达为3例（5.45%）。18例尖锐湿疣冰冻标本经RT-PCR法检测,HPV6b和11型E7 mRNA阳性表达分别为15例和10例,双阳性表达7例,其阳性表达型别与免疫组化结果完全一致,符合率均为100%。 结论 该自行制备的HPV6b和11型E7蛋白多克隆抗体免疫组化法检测尖锐湿疣皮损,检测结果直观,可以观察到HPV6b和11型感染细胞在病损中的分布位置。     

包头地区尖锐湿疣的发病人群及HPV的型别分析

目的 分析包头地区尖锐湿疣（CA）的发病人群及其人工乳头瘤病毒（HPV）的型别。方法 用HPV通用型和特异型引物6／11、16／18、16、18型对CA组织进行聚合酶链反应（PCR）检测。结果 通用型100％、6／11型66．8％、16型1％、6／11＋16／18型13．3％,其它型10．5％。结论 包头地区CA感染以HPV6／11型为主。HPV感染高峰年龄是20－40岁。女性明显高于男性,尤其是生育期妇女。     

山东省潍坊市尖锐湿疣患儿乳头瘤病毒检测和型别分析

目的 检测山东省潍坊市尖锐湿疣患儿中人乳头瘤病毒（HPV）感染情况及基因亚型。 方法 采用基因扩增技术及导流杂交分型技术,对24例患儿疣体的HPV进行检测分型。 结果 24例尖锐湿疣检出23例HPV阳性,阳性率95.8% ,20例单一型HPV感染占87.0%,混合感染型3例占13.0%;感染型别依次是HPV6（9/23）,HPV16（6/23）,HPV11（5/23）,HPV58（3/23）,HPV18（2/23）,HPV53（1/23）。 结论 该地区尖锐湿疣患儿HPV型别以6、16、11、58、18为主,患儿感染HPV型别与生活中密切接触者感染的型别相似。     

尖锐湿疣患者人乳头瘤病毒11型L1基因的克隆及序列分析

目的克隆和分析HPV11的晚期表达基因序列L1及其编码的氨基酸序列,为HPV感染的检测和基因工程疫苗研制提供基础。方法采用PCR技术扩增了1例尖锐湿疣患者疣组织中HPV 11 L1基因,并对其进行了克隆及全序列分析。结果本例尖锐湿疣临床标本HPV 11 L1基因与GenBank收录的原型(收录号:M14119)比较有8个碱基变异。结论 中国地区尖锐湿疣HPV 11 L1基因存在着变异。     

上海地区女性外阴尖锐湿疣患者宫颈HPV感染及分型研究

目的 探讨上海地区女性外阴尖锐湿疣患者的宫颈HPV感染情况.方法 采用美国FDA批准的捕获杂交试验和基因芯片技术对194例女性外阴尖锐湿疣患者皮损和宫颈脱落细胞进行HPV基因型别研究.结果 194例女性尖锐湿疣患者外阴皮损HPV检出率为100%,其中低危型HPV 144例占74.2%,高危型HPV 50例占25.8%.低危型中136例为单一型别感染,占94.4%,均为HPV6或HPV11;高危型中43例为低危与高危型HPV混合感染,占86%.194例患者中,166例(85.6%)合并宫颈HPV感染,其中低危型感染者119例,感染率为61.4%,高危型感染46例,感染率为23.7%.外阴和宫颈皮损HPV型别的一致性达到95.8%(159/166),低危型HPV感染率从高到低依次为HPV11＞HPV6＞HPV53,高危型依次为HPV16＞HPV18＞HPV52＞HPV31＞HPV45、HPV58.结论 上海地区女性外阴尖锐湿疣患者中高危型HPV感染率较高,且近1/4患者合并宫颈高危型HPV感染.     

The prevalence of "high-risk" HPV types in penile condyloma-like lesions: correlation between HPV type and morphology.

OBJECTIVE--To evaluate the prevalence of "high-risk" human papilloma virus (HPV) types in penile condyloma-like lesions and to correlate HPV types with clinical and histological features. DESIGN--The study included 94 male patients with signs of penile HPV infection. From acuminate, papular and macular lesions, specimens were collected for HPV DNA hybridisation, using the dot blot and Southern blot techniques. Biopsy specimens from 51 cases were examined by light microscopy for signs of koilocytosis and dysplasia. SETTING--The STD outpatient clinic of the Department of Dermatovenereology of Sahlgrenska Hospital, Göteborg, Sweden. RESULTS--In 79 (90%) of 88 patients HPV DNA was detected by dot blot. Of 51 cases examined by histology 88% disclosed an evident koilocytosis. "High-risk" HPV types (16, 18, 31, 33, 35) were demonstrated in 8% of acuminate, 24% of papular and 56% of macular lesions. In 29% of 51 lesions examined histologically moderate to severe dysplasia was observed. There was a significant correlation between "high-risk" HPV types and dysplasia. CONCLUSION--"High-risk" HPV types are prevalent in papular and especially macular penile condyloma-like lesions. The histological finding of koilocytosis concomitant with dysplasia strongly indicates infection with a "high-risk" HPV type. Although the risk of penile cancer is low, it is from an epidemiological point of view important to diagnose these lesions. Until simple tests for HPV typing are available, biopsy for light microscopy (histology) should be obtained liberally from papular and macular condyloma-like lesions. In atypical cases of balanoposthitis HPV aetiology should also be considered.     

聚合酶链反应及转印杂交对尖锐湿疣复发的病因学研究

用通用引物介导的聚合酶链反应及其产物斑点杂交技术，对１５例复发及相应的原发尖锐湿疣（ＣＡ）皮损中人乳头瘤病毒相关序列进行了检测和分型；同时还对３５例原发ＣＡ周围外观正常皮肤组织ＤＮＡ作了转印杂交分析。结果表明隐性感染是ＣＡ复发的非常重要原因，再感染也是ＣＡ复发不容忽视的原因。     

尖锐湿疣人乳头瘤病毒的基因分型

目的：检测尖锐湿疣中人乳头瘤病毒（HPV）的基因型别。方法：用通用引物进行聚合酶链反应,扩增尖锐湿疣中HPV DNA,用生物素标记的特异性基因分型探针,对聚合酶链反应扩增后的HPV DNA进行斑点杂交,结果：50例尖锐湿疣标本经PCR扩增后48例HPV DNA阳性,阳性标本中HPV6,11,16,18型的检出率分别为16．7％（8／48）,37．5％（18／48％）,14．6％（7／48）,4．2％（2／48）,多重型别检出率为22．9％（11／48）,结论 HPV11型感染是尖锐湿疣发病的首要原因。     

巨大尖锐湿疣患者人乳头瘤病毒型别检测

目的 探讨巨大尖锐湿疣患者皮损人乳头瘤病毒（HPV）-DNA型别。方法 收集门诊2007年1月至2010年1月巨大尖锐湿疣患者67例,凯普导流杂交基因芯片技术对疣体行HPV-DNA分型检测。结果 67例巨大尖锐湿疣患者中63例检测出HPV-DNA,阳性率为94.02%。HPV-DNA阳性者中检出低危型HPV 84例次（占60.87%）,高危型HPV 54例次（占39.13%）。HPV亚型仍以低危型HPV6、11为主,但高危型HPV16、18偏高。结论 巨大尖锐湿疣感染高危型HPV16、18偏高。     

Anal condylomas in men. 1. Histopathological and virological assessment.

A series of 128 biopsy specimens from anal condylomas in 73 homosexual or bisexual and 38 heterosexual men (mean (SD) age 31.8 (9.6) years) were subjected to histological assessment and human papillomavirus (HPV) typing by in situ DNA hybridisation with 35S-labelled HPV 6, 11, 16, 18, 31, and 33 probes. Most patients were also tested serologically for antibodies to human immunodeficiency virus (HIV). As evaluated on light microscopy, most (74%, 95/128) of the lesions were exophytic (papillary) acuminate warts, 15% (19) were flat, and 11% (14) were pigmented papulous lesions. No signs of anal intraepithelial neoplasia (AIN) were seen in 70% (90) of the 128 biopsy specimens (NAIN), 27% (35) were classified as showing AIN I, and another 2% (three) as AIN II. AIN was significantly (p less than 0.05) more often associated with papulous lesions, only 43% (6/14) of which showed NAIN compared with 72% (68/98) of acuminate condylomas. The duration of disease was directly related to the presence and severity of AIN in the lesions; thus in 47 lesions that had been present for more than 12 months, NAIN was found in 31 (66%), AIN I in 14 (30%), and AIN II in two (4%). HPV DNA of at least one of the six types tested for was detected in 109/125 (87%) lesions. HPV 6 and HPV 11 were the two most common types, comprising 57% (62) and 37% (40), respectively, of the 109 HPV DNA positive cases. Only seven (6%) biopsy specimens were associated with any of HPV types 16, 18, 31, or 33, which carry a high risk of potential malignant transformation. No association was found between sexual preferences of patients and the incidence of any of the various HPV types. Neither did the distribution of the various HPV types differ between men with antibody to HIV and those without antibody. All the men with antibody to HIV were homosexual or bisexual. On microscopy, 93% (38) of 41 lesions containing HPV 11 and 75% (48/64) of HPV 6 lesions were of the acuminate wart type; in comparison, the remaining 16 HPV 6 lesions were equally either flat or papulous (eight, 13% each). Of the 64 HPV 6 and 41 HPV 11 associated lesions, 73% (47) and 63% (26), respectively, were classified as NAIN. Only two lesions were associated with HPV 16, and both showed mild dysplasia. On the other hand, two HPV 6 induced lesions were associated with AIN II. No differences were found between HPV 6 and HPV 11 in duration of disease; (39%, and 27% respectively, had been present for more than 12 months). The results showed that overt anal wart disease was associated with HPV types 6 and 11 in most cases. Although HPV types considered as being of higher oncogenic potential were detected relatively rarely, the associated AIN in a relatively high proportion (31% 32/105) of HPV 6 or 11 induced lesions indicated that a malignant potential, even for HPV 11 associated anal warts, cannot be excluded.     

热休克蛋白在尖锐湿疣组织中的检测及其与HPV感染相关性

目的　探讨热休克蛋白 70(HSP70)在尖锐湿疣(CA)组织中的检测及其与人乳头瘤病毒 (HPV)感染的关系。方法　采用免疫组化法检测了 24例CA、46例宫颈癌和 28例正常宫颈组织HSP70,用PCR技术对各组织标本的HPV-DNA进行检测和 6, 1 1, 1 6, 1 8分型。结果　HSP7 0在CA和宫颈癌组的阳性率显著高于正常宫颈对照组 (P<0. 01)。CA组HPV6, 11型DNA阳性者HSP70的检出率显著高于阴性者(P<0. 05),宫颈癌组HPV16, 18型DNA阳性者与阴性者HSP70的检出率差异无显著性(P>0. 05)。结论　HSP70的表达与尖锐湿疣的发病有关,HSP70在尖锐湿疣组织中的表达与HPV的感染明显相关。     

宫颈尖锐湿疣的分化程度与HPV亚型的关系

为探讨宫颈尖锐湿疣的分化程度与HPV感染亚型的关系,用PCR方法分析宫颈尖锐湿疣的HPV亚型,电镜观察宫颈尖锐湿疣的超微病理变化,免疫组化方法检测宫颈尖锐湿疣中HPV抗原表达。结果显示,扁平型宫颈尖锐湿疣HPV6/11的感染率明显高于HPV16/18的感染率(P<0.01),乳头型宫颈尖锐湿疣HPV16/18的感染率明显高于HPV6/11的感染率(P<0.01);扁平型宫颈尖锐湿疣主要表现为凹空细胞,乳头型宫颈尖锐湿疣主要表现为细胞核的病理性改变;具有凹空细胞的宫颈尖锐湿疣中,HPV6/11的阳性率明显高于HPV16/18的阳性率(P<0.01),以细胞核病理性改变为特征的宫颈尖锐湿疣中,HPV16/18的阳性率明显高于HPV6/11的阳性率(P<0.01);HPV抗原的表达仅见于HPV6/11感染且凹空细胞明显的宫颈尖锐湿疣。结果表明,宫颈尖锐湿疣的分化程度与HPV感染亚型有关。     

尖锐湿疣凹空细胞的观察

本文对９７例尖锐湿疣（ＣＡ）进行聚合酶链反应检测人乳头瘤病毒（ＨＰＶ）ＤＮＡ及光镜下观察凹空细胞的特征。结果显示，９５例有凹空细胞，占９７．９４％。凹空细胞多分布在棘层的中、上层。根据核的改变将凹空细胞分为九型，以镰刀形核、毛虫样核和核大浓染型出现例数最多，分别为７５／９５、７４／９５和６１／９５，此三型对ＣＡ最有诊断价值。ＨＰＶ６、１１阳性９３例（９５．８８％），ＨＰＶ１６、１８、３１、３３、５２ｂ、５８阳性２例（２．０６％），ＨＰＶ６、１１、１６、１８、３１、３３、５２ｂ、５８阴性２例（２．０６％），但有典型的凹空细胞。２例ＣＡ无凹空细胞而ＨＰＶ６、１１为阳性。