尖锐湿疣的临床与实验研究

对本科性接触传播疾病（ＳＴＤ）专科门诊１９８９￣１９９６年间的各种ＳＴＤ数据统计发现,尖锐湿疣痊居各种性传播疾病病例数的第一位或第二位,用ＰＣＲ和斑点杂交等方面检测尖锐湿疣组织标本中ＨＰＶ ＤＮＡ,表明ＰＣＲ方法更加敏感。用ＰＣＲ方法检测宫颈鳞癌及宫颈和外阴尖锐湿疠ＨＰＶ感染,证实宫颈鳞癌以高危型ＨＰＶ１６为主,而外阴及宫颈尖锐湿疣均以低危型ＨＰＶ６／１１为主。为了鉴别尖锐湿疣与假性湿疣,进行了组     

女性尖锐湿疣和假性湿疣的HPV原位杂交检测与病理相关研究

目的 从组织学角度结合ＨＰＶ原位杂交检测探讨尖锐湿疣和假笥湿疣及相关病变的临床病理特点。方法 收集１９６例活检标本,随机抽取典型尖锐湿疣７０例,假性湿疣３０例,相关病变１０例作ＨＰＶ６Ｂ、１原位杂交检测分析,观察ＨＥ组织切片。结果 本组材料中,７０例（１００％）典型尖锐湿疣和３例（１０％）假性湿疣ＨＰＶ表达阳性,相关病变均表达阴性,尖锐湿疣和相关病变ＨＰＶ表达具有显著差异性（Ｐ〈０．００１）。结论     

多对引物PCR方法鉴别尖锐湿疣和假性湿疣

应用多对引物ＰＣＲ方法对尖锐湿疣９５例和假性湿疣３３例进行了鉴别。结果９５例尖锐湿疣中８６例（９０．５％）ＨＰＶ６／１１ＤＮＡ阳性，３３例假性湿疣ＨＰＶＤＮＡ阴性，两组有非常显著差别（Ｐ＜０．００１）。本研究结果显示假性湿疣与ＨＰＶ无关。ＰＣＲ方法准确、省时、方便，是鉴别尖锐湿疣与假性湿疣的理想方法。     

尖锐湿疣皮损人乳头瘤病毒基因分型的研究

目的探讨外生殖器及子宫颈尖锐湿疣患者皮损脱落细胞或组织中人乳头瘤病毒(human papillomavirus, HPV)11种基因型别的分布状况及临床意义.方法取子宫颈及生殖器尖锐湿疣疣体脱落细胞或尖锐湿疣组织,经PCR扩增后其产物与11种常见HPV亚型探针进行导流快速反斑点印迹杂交.结果 132例尖锐湿疣患者标本中,经PCR扩增并通过通用探针杂交分析后,有123例标本HPV DNA阳性93%.PCR阳性标本中,112例成功分型.高危型HPV16、18、31、33、52、58和66分别检出42例(31.8%)、22例(16.7%)、9 例(6.8%)、6例(4.5%)、18例(13.6%)、17例(12.9%)和5例(3.8%).低危型HPV6、11、53型分别检出55例(41.7%)、75例(56.8%)和7例(5.3%).危险度待定HPV亚型CP8304检出率为4.5%(6/132).有63.9%患者存在2～5型、高危/低危型HPV交叉感染,二重、三重、四重和五重型别感染检出率分别为31.8%(42/132)、26.5%(35/132)、4.5%(6/132)和1.5%(2/132).结论 HPV感染趋于多重化;HPV 11、6、16型感染仍然是尖锐湿疣发病的主要型别.     

女阴假性湿疣病因及诊断的研究

为了明确女阴假性湿疣（ＰＣＶ）的病因及与尖锐湿疣（ＣＡ）有无相关性，本文采用聚合酶链式反应（ＰＣＲ）技术以及病理、电镜等方法对４２例ＰＣＶ的病因及诊断进行了研究。结果表明，ＰＣＶ与人乳头瘤病毒感染无关，与长期慢性非特异性刺激有一定关系，与念珠菌感染关系不密切；ＰＣＲ是诊断ＰＣＶ的一种快速、灵敏，特异性强的方法。     

COMPARISON BETWEEN IMMUNOFLUORESCENCE AND PCR IN DETECTING HUMAN PAPILLOMA VIRUS IN CONDYLOMA ACUMINATA

CONDYLOMA acuminata (CA) was the secondmost common sexually transmitted disease in Chi-na1caused by human papilloma virus (HPV)·Polymerase chain reaction (PCR) is the most widely usedtechnique to detect HPVs in CA and in the smear of cer-vix·2Immnofluor…     

假性湿疣样尖锐湿疣的病原学研究

目的 研究临床酷似假性湿疣的尖锐湿疣的病原学。 方法 回顾性分析温州医科大学附属第一医院2013年1月-2015年12月收治的典型尖锐湿疣(15例)、假性湿疣(15例)、假性湿疣样尖锐湿疣(25例)患者的临床资料。对比分析3组患者的临床表现、病理学表现及HPV分型,以探讨临床酷似假性湿疣的尖锐湿疣的病原学特征。 结果 典型湿疣皮损表现为疣状粗糙、易破溃的淡红色小丘疹;假性湿疣皮损多为发于两侧小阴唇内侧的1~2 mm大小群集丘疹;假性湿疣样尖锐湿疣临床表现酷似假性湿疣。典型尖锐湿疣醋酸白实验阳性,假性湿疣阴性,假性湿疣样尖锐湿疣弱阳性。尖锐湿疣HPV全部阳性[4种低危HPV(6、11、43、55)、10种高危HPV(16、18、33、39、51、52、53、58、59、66)],1种HPV亚型4例,2种HPV亚型6例,3种HPV亚型3例,4种HPV亚型2例;假性湿疣HPV检测全部阴性;假性湿疣样尖锐湿疣HPV全部阳性[3种低危HPV(6、11、81)、7种高危型HPV(16、18、33、35、51、53、58)],1种HPV亚型8例,2种HPV亚型12例,3种HPV亚型3例,4种HPV亚型2例。 结论 临床表现为假性湿疣的患者中可能包含高危HPV感染的尖锐湿疣。      

PCR用于尖锐湿疣诊断的价值

用人乳头瘤病毒基因Ｌ１区通用引物（ＭＹ１１／ＭＹ０ｇ）进行聚合酶链反应，检测５０例尖锐湿疣新鲜组织。其中含３例病变不典型病理活组织检查不能确诊者。ＨＰＶ，ＤＮＡ阳性率１００％ＨＰＶ－６．１１，１６，１８型分别为３６％，７０％，１４％．８％。结果显示：（１）ＰＣＲ检测ＨＰＶ具有灵敏。特异、简单快速等优点，可有助于不典型尖锐湿疣患考的诊断，（２）ＭＹ１１／ＭＹ０９在筛选ＨＰＶ感染时是较理想的引物，（３）尖锐湿疣组织中以ＨＰＶ６、１１为最常见型别。     

Detection and typing of human papillomavirus infection of uterine cervix in young women by non-isotopic subgenomic probes on Southern blot--a report of studies in Sweden

Human papillomavirus (HPV) was detected by non-isotopic subgenomic probes on Southern blot hybridization (Oncor Inc., Gaithenberg, MD). The 108 samples which were obtained from patients below the age of 30 (mean age 19.0 +/- 2.8 yrs). All samples were collected from the ectocervical mucosa by Cytobrush (Medscand, Malm?, Sweden) as previously described. HPV-DNA was detected in 16 cases (15%). In 36 cases (33%) the patients presented clinical findings of condyloma or cervical intra-epithelial neoplasia (CIN) at simultaneous vaginal ectocervical smears. HPV-DNA was detected in 12 out of 36 cases. On the other hand, in the patients without subclinical findings HPV-DNA were detected in only 4 out of 70 cases (6%). HPV-DNA was detected in one of the 4 patients with accuminate condyloma of the cervix and in 4 out of 16 cases with similar lesions in the vulva. The only HPV type found in these patients was type 6. Papillomavirus was also detected in 5 out of 16 cases (31%) with flat condylomas, most studied types being represented here. CIN was only reported two patients both of them carrying HPV-DNA (types 16 and 18 respectively). The most common type of virus was HPV 6. Combined infections with two or three types were seen in 5 out of 16 HPV-positive cases (31%). Such cases are readily detected and typed with the present Southern blot procedure, where the use of subgenomic probes enables the distinction of all types, even when present in the same sample.     

HPV Infection and Distribution of HPV Genotypes on Patients with Condyloma Acuminatum

Objective To determine the incidence of infection with HPV and the distribution of HPV genotypes on patients with Condyloma acuminatum. Methods Twenty-three different HPV types were detected by PCR and reverse dot blot （RDB） hybridization over all 6 508 samples of vulva and cervix uteri in patients with condyloma acuminatum. Including 18 types were high-risk （HR）-HPV （16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 83 and MM4）, and 5 types were low-risk （LR）-HPV （6, 11, 42, 43 and 44）. Results Among 6 508 cases, there were 3 288 cases with HPV infection and the incidence rate was 50.52%. The positive HPV patients overlap all 23 genotypes detected. There were 2 038 cases infected with a single HPV type and 1 250 cases infected with multiple HPV types. The constituent ratios in positive cases were 61.98% and 38.02%, respectively. There were 1 453 cases only with LR-HPV types infection, 945 cases only with HR-HPV types and 890 cases both with LR-HPV and HR-HPV infection, and the constituent ratios were 44.19%, 28.74% and 27.07%. There were 4 843 times positive HPV infection, including HR-HPV 2 361 times and LR-HPV 2 482 times. The common HR-HPV genotypes were 16, 52, 58, 56, 18, 66 and 33, and the incidence ratios were 6.31%, 5.06%, 4.04%, 2.60%, 2.41%, 2.40% and 2.28%. And the common LR-HPV genotypes were 6, 11 and 43, and the incidence ratios were 16.98%,11.09% and 6.75%, respectively. Overlap 23 types, the most common geno- types were HPV6 and HPVll, the incidence rates were higher than others （P〈0.05）. Conclusion HPV infection, especially with HR-HPV genotypes infect genital tract, which caused Condyloma acuminatum and cervical lesions, or cervical cancer. The detection of HPV genotypes was very important to prevent, diagnose early and therapy for cervical lesions or cervical cancer.     

三种方法诊断尖锐湿疣的对比研究

目的探讨原位分子杂交法诊断尖锐湿疣的敏感性与特异性，并与组织病理及醋白试验相比较。方法应用原位分子杂交法[所用探针为地高辛标记的DNA探针（HPV6／11，HPVl6／18，HPV31／33）]、组织病理及醋白试验分别检测实验组和对照组标本。结果37例标本中，醋白试验34例阳性；组织病理检查37例均阳性；原位分子杂交法的总阳性率为94．6％，HPV6／11阳性31例（阳性率83．7％），HPV16／18阳性2例（阳性率5．4％），1例标本HPV6／11及HPVl6／18均阳性（2．7％），HPV31／33阳性1例（阳性率2．7％），阳性标本可见细胞核着蓝色，阳性反应物主要分布在棘层空泡化细胞的细胞核内。结论原位分子杂交法检测HPV敏感性高，特异性强，能对感染有明确的组织学定位，而且还能进一步鉴定感染的型别，因此是诊断和研究尖锐湿疣的较好和先进方法。     

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

Objective To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immunogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic expression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunogenicity study. Results The recombinant plasmid (pcDNA3. 1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice‘s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata.     

人乳头瘤病毒DNA阳性的外阴,宫颈组织病理学表现

目的 探讨外阴、宫颈人乳头瘤病毒（ＨＰＶ）感染与局部组织病理学改变的关系,评价组织病理学检查在ＨＰＶ感染诊断中的意义。方法 采用原位杂交和聚合酶链反应（ＰＣＲ）方法,对１１２例外阴、宫颈病变进行了ＨＰＶ６Ｂ／１１、１６、１８型ＤＮＡ检测,并对ＨＰＶ ＤＮＡ阳性的病变进行组织病理诊断。结果 ＨＰＶ ＤＮＡ阳性者８２例,占７３．２１％,８２例中,组织病理诊断为尖锐湿疣者２２例,其余６０例组织病理学均无     

用生物素标记的DNA探针检测尖锐湿疣和宫颈癌中的人类乳头瘤病毒DNA序列

用生物素标记的HPV6/11DNA和HPV16/18DNA探针,对尖锐湿疣和宫颈癌标本进行原位杂交。结果大多数尖锐湿疣含HPV6/11DNA,所以尖锐湿疣的主要致病因子是H2V6/11.HPV6/11DNA阳性细胞主要位于上皮的表层,少数位于深层。个别合并HPV16/18感染,或仅为HPV16/18所感染的病人恶变的可能性大。大多数宫颈癌病人含HPV16/18DNA,提示宫颈癌的主要致病因子为HPV16/18。一些病人合并HPV6/11感染。在未找见HPV16/18DNA的病例中,可能存在其它致癌因子。     

宫颈癌组织中人乳头状瘤病毒16型DNA序列的检测

应用DNA斑点分子杂交技术检测了54例宫颈癌组织中人乳头状瘤病毒16型DNA序列,并应用DNA Southern印迹杂交技术分析了部分斑点杂交阳性的标本。结果表明,50%的宫颈癌标本中HPV16DNA阳性,而在18例正常宫颈组织中未测到HPV16DNA序列;Southern印迹杂交发现HPV16DNA以整合状态存在。提示HPV16与宫颈癌的发生密切相关。     

用生物素标记的DNA探针检测尖锐湿疣和宫颈癌中的人类乳头瘤病毒DNA序列

用生物素标记的HPV6/11DNA和HPV16/18DNA探针,对尖锐湿疣和宫颈癌标本进行原位杂交。结果大多数尖锐湿疣含HPV6/11DNA,所以尖锐湿疣的主要致病因子是H2V6/11.HPV6/11DNA阳性细胞主要位于上皮的表层,少数位于深层。个别合并HPV16/18感染,或仅为HPV16/18所感染的病人恶变的可能性大。大多数宫颈癌病人含HPV16/18DNA,提示宫颈癌的主要致病因子为HPV16/18。一些病人合并HPV6/11感染。在未找见HPV16/18DNA的病例中,可能存在其它致癌因子。     

应用PCR技术检测尖锐湿疣患者HPV感染

取尖锐湿疣的３０例患者外阴湿疣标枉和宫颈分泌物及同期非尖锐湿疣患者宫艰苦泌物，用聚合酶链反应技术检测其中ＨＰＶ６．１１ＤＮＡ和ＨＰＶ１６．１８ＤＮＡ。结果表明，尖锐湿疣患者宫颈人乳头瘤病毒检出率显著高于非尖锐湿疣患者，以ＨＰＶ６．１１为主，ＨＰＶ１６．１８在二者之间无差别。提示外阴尖锐湿疣主要由ＨＰＶ６．１１感染引起，应用ＰＣＲ技术检测湿疣组织中ＨＰＶ ＤＮＡ可为痢疾诊断提供客观依据。     

聚合酶链反应技术检测女性生殖道尖锐湿疣的临床应用

目的：早期发现亚临床和潜伏性人乳头瘤病毒（HPV）感染,给予及时处理,控制病情向尖锐湿疣临床期发展,方法：应用聚合酶链反应（PCR）技术检测可疑女性生殖道HPV感染,并通过活组织检查予以验证。结果：可疑女性生殖道尖锐湿疣 56例,聚合酶链反应检测HPV（6,11型）感染阳性53例,添组织检查验证为尖锐疣48例,PCR检测HPV感染的敏感性为1005,准确性为85．75。结论应用PCR技术检测HPV（6,11型）能发现亚临床表现的女性生殖道尖锐湿疣和HPV感染携带者。     

宫颈尖锐湿疣组织中HPV分型与端粒酶活性的研究

目的由于宫颈尖锐湿疣(CA)皮损中HPV高危型感染常与其癌变有关，而端粒酶活性增高多见于恶性疾病。该研究探讨宫颈CA皮损中HPV分型与端粒酶活性的关系。方法采用荧光定量PCR检测HPV病毒分型，端粒重复序列扩增文件-酶标法(TRAP—ELISA)检测端粒酶活性。结果尖锐湿疣患者皮损中端粒酶活性明显高于正常皮肤组织；24例(40．0％)HPV16／18型阳性，34例(56．7％)HPV6／11型阳性，2例未检测到HPV；HPV16／18型感染的皮损中端粒酶活性明显高于HPV6／11型感染者。结论认为端粒酶活性与HPV型别有关，可能与HPV病毒协同参与CA发病，HPV16／18型阳性者在临床可能有癌变倾向，抑制端粒酶活性可能可以阻碍疣体生长，为临床治疗提供一个新的方向。