特殊皮肤疣中人乳头瘤病毒感染状况的临床特征和基因分型研究
摘要
疣是人乳头瘤病毒(HPV)感染所致常见的皮肤科疾病之一,在临床工作中,常遇到如下困惑:发生在指(趾)间、腋下、腹股沟或口腔内形态酷似尖锐湿疣的皮疹如何诊断?发生在肛门、外生殖器的类似寻常疣且数年无变化的皮疹是否也诊断为尖锐湿疣?其HPV型别与寻常疣相同还是与尖锐湿疣相同?非生殖器部位皮损是否存在高危型HPV感染的可能?继发肿瘤的机会有多大?本研究拟通过了解此类“特殊皮肤疣”的形态、病理、HPV型别之间的相互关系,旨在回答临床工作中遇到的问题,寻找其分子生物学、临床治疗学和流行病学资料,为这组“特殊皮肤疣”诊治提供相应的理论基础,为预后提供一定的指导作用,并为进一步的深入研究奠定基础。全文具体分为三部分。
第一部分“特殊皮肤疣”中人乳头瘤病毒的基因分型研究
目的探讨特殊部位(趾间,腹股沟)皮肤疣、特殊形态(鸡冠状、菜花状)头面部疣和特殊形态(乳头状、丘疹状)生殖器疣等四组特殊皮肤疣的HPV基因型别分布。
方法通过直接测序法对“特殊皮肤疣”中的HPV进行基因分型研究。根据测序所得到的型别,结合文献中皮肤疣的常见型别,分别设计型特异引物,通过型特异性PCR来验证和补充测序结果。
结果(1)趾间组20例、腹股沟组29例,HPV型别以HPV7为主;头面部组30例HPV型别以HPV27、7为主;生殖器组18例HPV型别以HPV6、11为主,未检测到HPV7;HPV2、7、27、57是寻常疣组中HPV主要的型别,腹股沟组和生殖器组均出现高危型(HPV16)。与寻常疣对照组相比,趾间组、腹股沟组和生殖器组HPV7构成比差异有显著统计学意义(P值分别为0.000、0.000和0.011);而头面部组HPV7构成比差异无统计学意义(P=0.413)。(2)7对TSP-PCR的结果表明,在趾间、腹股沟、生殖器组均存在多种型别的混合感染。两种方法的型别符合率较高(80～99%)。
结论趾间、腹股沟等皮肤皱褶处疣中HPV7的感染性较高,皮疹形态以菜花状、丘疹状为主。头面部菜花状疣HPV型别以HPV27、7为主,生殖器丘疹状疣HPV型别以HPV6、11为主,腹股沟疣和生殖器丘疹状疣中存在高危型HPV(HPV16)。各组均存在HPV混合型别感染。
第二部分特殊皮肤疣的临床病理特征研究
目的探讨“特殊皮肤疣”的临床和病理特征及各观察组的HPV基因型别与主要的临床病理特征之间的相关性。
方法对趾间疣、腹股沟疣、头面部疣和生殖器疣四组“特殊”皮肤疣的临床和病理特征进行归纳和总结,对各观察组的HPV基因型别与主要的临床病理特征之间的相关性进行统计学分析。
结果趾间组、腹股沟组患者均为男性,形态以菜花状、丘疹状为主,与寻常疣组形态相比,差异有统计学意义(菜花状P=0.011,丘疹状p=0.005)。疣体形态与HPV型别之间没有显著的关联性。共有56例患者的皮损作普通病理,表现寻常疣模式者18例,尖锐湿疣模式者16例,不易鉴别者22例。HPV7阳性(18份)的病理象特征:(1)乳头瘤样增生和棘层肥厚;(2)内含中等大小的透明角质颗粒的细胞深染;(3)显著的角化过度和角化不全。
结论HPV型别与疣体形态之间的相关性无显著的统计学意义。特殊皮肤疣病理可表现为典型的寻常疣病理像或尖锐湿疣病理像,HPV7阳性的病理像具有特征性组织学改变。结合特殊皮肤疣中HPV型别分布,建议特殊部位(趾间,腹股沟)皮肤疣、特殊形态(鸡冠状、菜花状)头面部疣诊断为寻常疣;特殊形态(乳头状、丘疹状)生殖器疣诊断为尖锐湿疣。
第三部分PCR产物的TA克隆及趾间皮屑HPV DNA的检测
目的对趾间疣组中的基因型进行进一步验证,初步探索HPV7在趾间疣中的寄居或致病状态。
方法采用PCR产物的TA克隆法对趾间疣组中的基因型进行进一步验证,设计趾间疣的病变阴性对照,取足癣患者和健康人群的趾间皮屑筛查有无HPV及HPV7的寄居。
结果取趾间疣组12份DNA标本,TA克隆其通用引物PCR产物,每一份标本的4个TA克隆子中至少有1个成功合成质粒重组体,其测序所得序列与直接测序法所得的序列序均有90%以上的一致性。共采集趾间糜烂型足癣患者皮损处皮屑53例作为阴性对照和健康人群趾间皮屑48例作为空白对照,HPV的阳性率分别为(15.09%,8/53;16.67%,8/48),未检测到HPV7 DNA。
结论测序型别可能是趾间疣的致病的HPV型别,趾间疣中所检测到的HPV7,可能处于致病状态,HPV7可感染免疫力正常的人群。
In dermatology clinical work,we encountered patients with warts in between toes,in armpits,on the groin,cauliflower-like warts on head and face;papule-like warts in genital site.These cauliflower-like lesions were similar to condylomata acuminata and papule-like lesions were similar to verruca vulgaris in morphology.What was the exact diagnosis with these special warts? Was there any high-risk genotypes in these warts?—We have studies on these special warts to expore basic molecular epidemiological and the clinical and histological characteristics to help dermatologist answer the stumbles in clinic.The paper consists of three parts.
Part 1 The distribution of the HPV genotype of human papillomavirus(HPV) infections among patients within special warts.
Objectives To investigate the molecular epidemiological characteristics of human papillomavirus(HPV) infections among patients with special warts (warts in webspace or groin;cauliflower-like warts on head and face; papule-like warts in genital site),analyze the distribution of the HPV genotype.
Methods DNA isolated from warts specimens were amplified by PCR and sequencing to identify HPV genotype.Type-special primer PCR(TSP-PCR) of HPV2,4,6,7,11,16,and 18 were also performed to compare the conformity of the two methods.
Results(1) Ninety-five cases were enrolled in the study groups.The main genotype was HPV7 in interdigital group(20 cases) and groin group(29 cases).In head and face group(30 cases),HPV7 and HPV27 were the main genotype.HPV6 and HPV11 were the main genotypes in genital group(18 cases). HPV2,7,27,57 were the main genotypes in verruca vulgaris group(31 cases). And a high-risk genotype(HPV16) was detected in genital and groin group. There was significant difference between some groups and the verruca vulgaris control group in constituent ratio of HPV7(interdigital group P=0.000,groin group P=0.000,genital group P=0.011).There was no significant difference between head and face group and verruca vulgaris group in constituent ratio of HPV 7(P=0.413).(2)The results of 7 pairs TSP-PCR showed that a variety of mixed-genotypes infection existed in all groups.The conincidence rates were 80-99%in sequencing and TSP-PCR.
Conclusions Warts located in webspace and groins are prone to be caused by HPV7.HPV7 and HPV27 are the main genotype in cauliflower-like warts on head and face.HPV6 and HPV11 the main genotypes in papule-like in genital warts.High risk HPV(HPV16) genetype DNA exists in genital and groin warts.
Part 2 Clinical and histological features and the potential relationship between HPV genotypes and clinical characteristics in patients with these special warts.
Objectives To expore the clinical and histological features and the potential relationship between HPV genotypes and clinical characteristics in patients with these special warts.
Methods The clinical and histological characteristics of those warts,the relationship between HPV genotypes and its clinical and histological characteristics were analyzed and statisticsed by SPSS.
Results The patients were all males,warts were mainly cauliflower-like and papule-like in interdigital group and groin group.There was significant difference between these two groups and the verruca vulgaris control group in gentile(P=0.000) and morphous(cauliflower-like,P=0.011;papule-like P=0.005).But there was no significant relationship between the morphology of the warts and HPV genotype.Fifty-six specimens were taken for analysis of histopathological patterns.It revealed 18 cases showing verruca vulgaris-like pathology patterns,16 condyloma accuminatum-like pathology features,and 22 cases could not determine verruca vulgaris-like or condyloma accuminatum-like pathology patterns.Main HPV7(9/18) positive samples' pathology manifestations matched the characteristics for HPV7 infection,which showed papillomatosis and acanthosis,deep-stained cells including keratohyalin granules in middle size,and obvious hyperkeratosis and parakertosis.
Conclusions Warts located in webspace and groins,mainly cauliflower-like and papule-like,are prone to offend male patients.But there is no significant relationship between the morphology and HPV type of the warts. Special warts could show verruca vulgaris-like or condyloma accuminatum-like pathology patterns.
Part 3 TA clone of PCR products and detection of the HPV DNA in scales of webspace.
Objectives To explore the role of HPV7 in the pathopoiesis of warts. Methods the gene sequence of HPV found in webspace specimens was studied with PCR and TA clone.The HPV and HPV7 DNA in scales of webspace in patients with tinea pedis and healthy persons were both detected by nest-PCR and TSP-PCR.
Results The purified and amplified DNA of 12 specimens in interdigital group were cloned with the pUC57-T Vector Cloning Kit.Four recombinant plasmids harboring inserts of the expected size were randomly selected and sequenced. The result showed the sequence shared 90%or more homology to the HPV genotype in direct sequencing.HPV DNA was detected in scales specimen of the webspace in 53 patients with tinea pedis and 48 healthy persons.HPV DNA positivity rate was 5/53 and 5/48 respectively in nest PCR.There was no HPV7 positive in all scale specimens in TSP-PCR.The results revealed statistically significant difference in sex ratio(P=0.002),and the positivity rate of HPV(P=0.000) or HPV7(P=0.000) between interdigital and tinea pedis groups or between interdigital and healthy person groups.There was no significant difference between the tinea pedis group and healthy persons group in sex ratio(P=0.835) or positivity rate in HPV7(P=0.829).
Conclusions HPV7 maybe plays a pathogenic role in the development of interdigital warts.It could induce warts in immunocompetent patients.
第一部分“特殊皮肤疣”中人乳头瘤病毒的基因分型研究
目的探讨特殊部位(趾间,腹股沟)皮肤疣、特殊形态(鸡冠状、菜花状)头面部疣和特殊形态(乳头状、丘疹状)生殖器疣等四组特殊皮肤疣的HPV基因型别分布。
方法通过直接测序法对“特殊皮肤疣”中的HPV进行基因分型研究。根据测序所得到的型别,结合文献中皮肤疣的常见型别,分别设计型特异引物,通过型特异性PCR来验证和补充测序结果。
结果(1)趾间组20例、腹股沟组29例,HPV型别以HPV7为主;头面部组30例HPV型别以HPV27、7为主;生殖器组18例HPV型别以HPV6、11为主,未检测到HPV7;HPV2、7、27、57是寻常疣组中HPV主要的型别,腹股沟组和生殖器组均出现高危型(HPV16)。与寻常疣对照组相比,趾间组、腹股沟组和生殖器组HPV7构成比差异有显著统计学意义(P值分别为0.000、0.000和0.011);而头面部组HPV7构成比差异无统计学意义(P=0.413)。(2)7对TSP-PCR的结果表明,在趾间、腹股沟、生殖器组均存在多种型别的混合感染。两种方法的型别符合率较高(80～99%)。
结论趾间、腹股沟等皮肤皱褶处疣中HPV7的感染性较高,皮疹形态以菜花状、丘疹状为主。头面部菜花状疣HPV型别以HPV27、7为主,生殖器丘疹状疣HPV型别以HPV6、11为主,腹股沟疣和生殖器丘疹状疣中存在高危型HPV(HPV16)。各组均存在HPV混合型别感染。
第二部分特殊皮肤疣的临床病理特征研究
目的探讨“特殊皮肤疣”的临床和病理特征及各观察组的HPV基因型别与主要的临床病理特征之间的相关性。
方法对趾间疣、腹股沟疣、头面部疣和生殖器疣四组“特殊”皮肤疣的临床和病理特征进行归纳和总结,对各观察组的HPV基因型别与主要的临床病理特征之间的相关性进行统计学分析。
结果趾间组、腹股沟组患者均为男性,形态以菜花状、丘疹状为主,与寻常疣组形态相比,差异有统计学意义(菜花状P=0.011,丘疹状p=0.005)。疣体形态与HPV型别之间没有显著的关联性。共有56例患者的皮损作普通病理,表现寻常疣模式者18例,尖锐湿疣模式者16例,不易鉴别者22例。HPV7阳性(18份)的病理象特征:(1)乳头瘤样增生和棘层肥厚;(2)内含中等大小的透明角质颗粒的细胞深染;(3)显著的角化过度和角化不全。
结论HPV型别与疣体形态之间的相关性无显著的统计学意义。特殊皮肤疣病理可表现为典型的寻常疣病理像或尖锐湿疣病理像,HPV7阳性的病理像具有特征性组织学改变。结合特殊皮肤疣中HPV型别分布,建议特殊部位(趾间,腹股沟)皮肤疣、特殊形态(鸡冠状、菜花状)头面部疣诊断为寻常疣;特殊形态(乳头状、丘疹状)生殖器疣诊断为尖锐湿疣。
第三部分PCR产物的TA克隆及趾间皮屑HPV DNA的检测
目的对趾间疣组中的基因型进行进一步验证,初步探索HPV7在趾间疣中的寄居或致病状态。
方法采用PCR产物的TA克隆法对趾间疣组中的基因型进行进一步验证,设计趾间疣的病变阴性对照,取足癣患者和健康人群的趾间皮屑筛查有无HPV及HPV7的寄居。
结果取趾间疣组12份DNA标本,TA克隆其通用引物PCR产物,每一份标本的4个TA克隆子中至少有1个成功合成质粒重组体,其测序所得序列与直接测序法所得的序列序均有90%以上的一致性。共采集趾间糜烂型足癣患者皮损处皮屑53例作为阴性对照和健康人群趾间皮屑48例作为空白对照,HPV的阳性率分别为(15.09%,8/53;16.67%,8/48),未检测到HPV7 DNA。
结论测序型别可能是趾间疣的致病的HPV型别,趾间疣中所检测到的HPV7,可能处于致病状态,HPV7可感染免疫力正常的人群。
In dermatology clinical work,we encountered patients with warts in between toes,in armpits,on the groin,cauliflower-like warts on head and face;papule-like warts in genital site.These cauliflower-like lesions were similar to condylomata acuminata and papule-like lesions were similar to verruca vulgaris in morphology.What was the exact diagnosis with these special warts? Was there any high-risk genotypes in these warts?—We have studies on these special warts to expore basic molecular epidemiological and the clinical and histological characteristics to help dermatologist answer the stumbles in clinic.The paper consists of three parts.
Part 1 The distribution of the HPV genotype of human papillomavirus(HPV) infections among patients within special warts.
Objectives To investigate the molecular epidemiological characteristics of human papillomavirus(HPV) infections among patients with special warts (warts in webspace or groin;cauliflower-like warts on head and face; papule-like warts in genital site),analyze the distribution of the HPV genotype.
Methods DNA isolated from warts specimens were amplified by PCR and sequencing to identify HPV genotype.Type-special primer PCR(TSP-PCR) of HPV2,4,6,7,11,16,and 18 were also performed to compare the conformity of the two methods.
Results(1) Ninety-five cases were enrolled in the study groups.The main genotype was HPV7 in interdigital group(20 cases) and groin group(29 cases).In head and face group(30 cases),HPV7 and HPV27 were the main genotype.HPV6 and HPV11 were the main genotypes in genital group(18 cases). HPV2,7,27,57 were the main genotypes in verruca vulgaris group(31 cases). And a high-risk genotype(HPV16) was detected in genital and groin group. There was significant difference between some groups and the verruca vulgaris control group in constituent ratio of HPV7(interdigital group P=0.000,groin group P=0.000,genital group P=0.011).There was no significant difference between head and face group and verruca vulgaris group in constituent ratio of HPV 7(P=0.413).(2)The results of 7 pairs TSP-PCR showed that a variety of mixed-genotypes infection existed in all groups.The conincidence rates were 80-99%in sequencing and TSP-PCR.
Conclusions Warts located in webspace and groins are prone to be caused by HPV7.HPV7 and HPV27 are the main genotype in cauliflower-like warts on head and face.HPV6 and HPV11 the main genotypes in papule-like in genital warts.High risk HPV(HPV16) genetype DNA exists in genital and groin warts.
Part 2 Clinical and histological features and the potential relationship between HPV genotypes and clinical characteristics in patients with these special warts.
Objectives To expore the clinical and histological features and the potential relationship between HPV genotypes and clinical characteristics in patients with these special warts.
Methods The clinical and histological characteristics of those warts,the relationship between HPV genotypes and its clinical and histological characteristics were analyzed and statisticsed by SPSS.
Results The patients were all males,warts were mainly cauliflower-like and papule-like in interdigital group and groin group.There was significant difference between these two groups and the verruca vulgaris control group in gentile(P=0.000) and morphous(cauliflower-like,P=0.011;papule-like P=0.005).But there was no significant relationship between the morphology of the warts and HPV genotype.Fifty-six specimens were taken for analysis of histopathological patterns.It revealed 18 cases showing verruca vulgaris-like pathology patterns,16 condyloma accuminatum-like pathology features,and 22 cases could not determine verruca vulgaris-like or condyloma accuminatum-like pathology patterns.Main HPV7(9/18) positive samples' pathology manifestations matched the characteristics for HPV7 infection,which showed papillomatosis and acanthosis,deep-stained cells including keratohyalin granules in middle size,and obvious hyperkeratosis and parakertosis.
Conclusions Warts located in webspace and groins,mainly cauliflower-like and papule-like,are prone to offend male patients.But there is no significant relationship between the morphology and HPV type of the warts. Special warts could show verruca vulgaris-like or condyloma accuminatum-like pathology patterns.
Part 3 TA clone of PCR products and detection of the HPV DNA in scales of webspace.
Objectives To explore the role of HPV7 in the pathopoiesis of warts. Methods the gene sequence of HPV found in webspace specimens was studied with PCR and TA clone.The HPV and HPV7 DNA in scales of webspace in patients with tinea pedis and healthy persons were both detected by nest-PCR and TSP-PCR.
Results The purified and amplified DNA of 12 specimens in interdigital group were cloned with the pUC57-T Vector Cloning Kit.Four recombinant plasmids harboring inserts of the expected size were randomly selected and sequenced. The result showed the sequence shared 90%or more homology to the HPV genotype in direct sequencing.HPV DNA was detected in scales specimen of the webspace in 53 patients with tinea pedis and 48 healthy persons.HPV DNA positivity rate was 5/53 and 5/48 respectively in nest PCR.There was no HPV7 positive in all scale specimens in TSP-PCR.The results revealed statistically significant difference in sex ratio(P=0.002),and the positivity rate of HPV(P=0.000) or HPV7(P=0.000) between interdigital and tinea pedis groups or between interdigital and healthy person groups.There was no significant difference between the tinea pedis group and healthy persons group in sex ratio(P=0.835) or positivity rate in HPV7(P=0.829).
Conclusions HPV7 maybe plays a pathogenic role in the development of interdigital warts.It could induce warts in immunocompetent patients.
引文
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[15]李泉,祖洪勇,杨清平,等.实时荧光聚合酶链反应技术与第2代杂交捕获法检测高危亚型人乳头瘤病毒的比较[J].重庆医学,2008,37(24):2802-2804.
[16]唐振华,李柱南,胡伟平.应用实时荧光聚合酶链反应技术与第二代杂交捕获法检测高危亚型人乳头瘤病毒[J].检验医学,2008,23(3):216-219.
[17]姚丽,张伟,王妹妹,等.液相芯片技术原理及应用简介[J].现代肿瘤医学,2008,16(12):2196-2198.
[18]刘思瑶,丁显平,朱一剑,等.液相芯片技术在人乳头瘤病毒检测和分型中的应用[J].四川大学学报自然科学版,2007,44(5):1111-1114.
[1]Astori G,Lavergne D,Benton C,et.al.Human papillomaviruses are commonly found in normal skin of immunocompetent hosts[J].J Invest Dermatol,1998,110(5):752-755.
[2]Berkhout RJ,Tieben LM,Smits HL,et.al.Nested PCR approach for detection and typing of epidermodysplasia verruciformis-associated human papillomavirus types in cutaneous cancers from renal transplant recipients[J].J Clin Microbiol,1995,33(3):690-695.
[3]de Villiers EM,Whitley C,Gunst K.Identification of new papillomavirus types[J].Methods Mol Med,2005,119:1-13.
[4]蒋明军.尖锐湿疣中人乳头瘤病毒感染状况的基因分型研究[D].北京:中国协和医科大学,2004:52-53.
[5]Takahashi S,Tsukamoto T.Condyloma acuminatum[J].Nippon Rinsho,2009,67(1):153-156.
[6]Kondo K.Development of an HPV vaccine-remaining issues and perspective[J].Nippon Rinsho,2009,67(1):62-68.
[7]Narisawa-Saito M,Kiyono T.Mechanisms of high-risk human papillomavirus-induced cervical carcinogenesis[J].Nippon Rinsho,2009,67(1):53-61.
[8]Thierry F.Transcriptional regulation of the papillomavirus oncogenes by cellular and viral transcription factors in cervical carcinoma[J].Virology,2009,20;384(2):375-379
[9]Zekri AR,Bahnassy AA,Seif-Eldin WM,et al.Role of human papilloma virus(HPV) in common and genital warts and its relation to P53expression[J].J Egypt Natl Canc Inst,2006,18(2):117-124.
[10]Oltersdorf T,Campo MS,Favre M,et al.Molecular cloning and characterization of human papillomavirus type 7 DNA[J].Virology,1986,149(2):247-250.
[11]Delius H.,Hofmann B.,Primer-directed sequencing of human papillomavirus types[J].Curr Top Microbiol.Immunol,1994,186:13-31.
[12]de Villiers EM,Neumann C,Oltersdorf T,et al.Butcher's wart virus (HPV 7) infections in non-butchers[J].J Invest Dermatol,1986,87(2):236-238.
[13]Greenspan D,de Villiers EM,Greenspan JS,et al.Unusual HPV types in oral warts in association with HIV infection[J].J Oral Pathol,1988,17(9-10):482-488.
[14]de Villiers EM.Prevalence of HPV 7 papillomas in the oral mucosa and facial skin of patients with human immunodeficiency virus[letter][J].Arch Dermatol,1989,125(11):1590.
[15]R(u|¨)dlinger R,Bunney MH,Grob R,et al.Warts in fish handlers[J].Br J Dermatol,1989,120(3):375-81.
[16]Meffert JJ,Anthony JS.Butcher' s warts:dermatological heritage or testable misinformation?[J].Arch Dermatol,2001,137(3):384-385.
[17]Lei YJ,Gao C,An R,Shi Q,et al.Development of a multiplex PCR method for detecting and typing human papillomaviruses in verrucae vulgaris[J].J Virol Methods,2008,147(1):72-77.
[18]Yan-JunLEI,Chen GAO,Chen WANG,et al.Molecular Epidemiological Study on Prevalence of Human Papillomaviruses in Patients with Common Warts in Beijing[J].Area iomedical and Environmental Sciences,2009,22(1):55-61.
[1]Jablonska S,Obalek S,Favre M,et al.The morphology of butchers' wans as related to papillomavirus types[J].Arch Dermatol Res,1987,279Suppl:66-72.
[2]de Villiers EM,Neumann C,Oltersdorf T,et al.Butcher's wart virus (HPV 7) infections in non-butchers[J].J Invest Dermatol,1986,87(2):236-238.
[3]赵辨.临床皮肤病学[M].北京:人民卫生出版社,2001:313.
[4]Aroni K,Lazaris AC,Ioakim-Liossi A,et al.Histological diagnosis of cutaneous "warty" carcinoma on a pre-existing HPV lesion[J].Acta Derm Venereol,2000,80(4):294-296.
[5]蒋明军,王书崎,龚向东.尖锐湿疣皮损中人乳头瘤病毒基因分型研究[J].中华皮肤科杂志,2005,38(5):262-264.
[6]Lu Y,Wang XL,Wu D,et al.Clinical features and epidemiological survey of perianal warts in 72 males[J].Zhonghua Nan Ke Xue,2006,12(10):923-926.
[7]Partridge JM,Koutsky LA.Genital human papillomavirus infection in men[J].Lancet Infect Dis,2006,6(1):21-31.
[8]Noel JC,Peny MO,Goldschmidt D,et al.Human papillomavirus type 1DNA in verrucous carcinoma of the leg[J].J Am Acad Dermatol,1993,29(6):1036-1038.
[9]Noel JC,Peny MO,Detremmerie O,et al.Demonstration of human papillomavirus type 2 in a verrucous carcinoma of the foot[J].Dermatology,1993,187(1):58-61.
[10]Bragg JW,Ratner D.Human papillomavirus type 2 in a squamous cell carcinoma of the finger[J].Dermatol Surg,2003,29(7):766-768.
[11]Chen AC,McMillan NA,Antonsson A.Human papillomavirus type spectrum in normal skin of individuals with or without a history of frequent sun exposure[J].J Gen Virol,2008,89(Pt 11):2891-2897.
[1]http://www.genscript.com/puc57,html
[2]于永利,麻彤辉.TA克隆及双链DNA:测序:介绍一种快速克隆及分析PCR产物的方法[J].中国免疫学杂志,1994,10(1):5-7.
[3]丁鹏,王家宁,黄永章,等.利用TA克隆载体构建pET15b-SOD1重组质粒[J].郧阳医学院学报,2005,24(2):65-68.
[4]de Villiers EM,Whitley C,Gunst K.Identification of new papillomavirus types[J].Methods Mol Med,2005,119:1-13.
[5]姚丽,张伟,王妹妹,等.液相芯片技术原理及应用简介[J].现代肿瘤医学,2008,16(12):2196-2198.
[6]刘思瑶,丁显平,朱一剑,等.液相芯片技术在人乳头瘤病毒检测和分型中的应用[J].四川大学学报自然科学版,2007,44(5):1111-1114
[7]Astori G,Lavergne D,Benton C,et.al.Human papillomaviruses are commonly found in normal skin of immunocompetent hosts[J].J Invest Dermatol,1998,110(5):752-755.
[8]Berkhout RJ,Tieben LM,Smits HL,et al.Nested PCR approach for detection and typing of epidermodysplasia verruciformis-associated human papillomavirus types in cutaneous cancers from renal transplant recipients[J].J Clin Microbiol,1995,33(3):690-695.
[9]孙海汐,蔡金挺,朱琳楠,等.一种改良的人DNA微量快速检验技术[J].中国实验诊断学,2007,11(6):788-790.
[10]陈辉,刘永波,谢庆瑞,等.有机酚法和Chelex-100法提取不同组织微量DNA 效果比较[J].郑州大学学报医学版,2004,39(6):988-991.
[11]张源明,钟良军,陈晓涛,等.Chelex-100法从颊粘膜拭子提取DNA进行基因型分析的研究[J].新疆医科大学学报,2003,26(5):430-431.
[12]童大跃,伍新尧,陆惠玲,等.DNA提取方法的比较及改良[J].中山大学学报医学科学版,2003,24.
[13]蒋明军.尖锐湿疣中人乳头瘤病毒感染状况的基因分型研究D.中国协和医科大学.2004:52-54.
[14]李远宏,陈光,张理涛,等.银屑病患者皮损及鳞屑中人乳头瘤病毒的检测[J].中华皮肤科杂志,2008,41(10):695-696.
[15]Alotaibi L,Provost N,Gagnon S,et al.Diversity of cutaneous human papillomavirus types in individuals with and without skin lesion[J].J Clin Virol,2006,36(2):133-140.
[16]Human papillomavirus:often harmless but in some cases carcinogenic [J].Prescrire Int,2007,16(89):115-119.
[1]de Villiers EM,Fauquet C,Broker TR,et al.Classification of papillomaviruses[J].Virology,2004,324(1):17-27.
[2]Dell G,Gaston K.Human papillomaviruses and their role in cervical cancer[J].Cell Mol Life Sci,2001,58(12-13):1923-1942.
[3]Oltersdorf T,Campo MS,Favre M,et al.Molecular cloning and characterization of human papillomavirus type 7 DNA[J].Virology,1986,149(2):247-250.
[4]DeliusH,Hofmann B.Primer-directed sequencing of human papillomavirus types[J].Curr Top Microbiol Immunol,1994,186:13-31.
[5]Keefe M,al-Ghamdi A,Coggon D,et al.Cutaneous warts in butchers [J].Br J Dermatol,1994,130(1):9-14.
[6]Keefe M,al-Ghamdi A,Coggon D,et al.Butchers' warts:no evidence for person to person transmission of HPV7.Br J Dermatol,1994,130(1):15-7.
[7]Rtldlinger R,Bunney MH,Grob R,et al.Warts in fish handlers [J].Br J Dermatol,1989,120(3):375-381.
[8]de Villiers EM,Neumann C,Oltersdorf T,et al.Butcher's wart virus (HPV 7)infections in non-butchers[J].J Invest Dermatol,1986,87(2):236-238.
[9]Greenspan D,de Villiers EM,Greenspan JS,et al.Unusual HPV types in oral warts in association with HIV infection[J].J Oral Pathol,1988,17(9-10):482-488.
[10]de Villiers EM.Prevalence of HPV 7 papillomas in the oral mucosa and facial skin of patients with human immunodeficiency virus [letter][J].Arch Dermatol,1989,125(11):1590.
[11]Meffert JJ,Anthony JS.Butcher' s warts:dermatological heritage or testable misinformation? [J].Arch Dermatol.2001,137(3):384-385.
[12]Majewski S,Jablonska S,Favre M,et al.Human papillomavirus type 7 and butcher's warts[J].Arch Dermatol,2001,137(12):1655-1656.
[13]Ritzkowsky A,Weissenborn S,Krieg T,et al.Extensive human papillomavirus type 7-associated orofacial warts in an immunocompetent patient[J].Acta Derm Venereol,2001,81(2):130-133.
[14]Jablonska S,Obalek S,Favre M,et al.The morphology of butchers' wans as related to papillomavirus types[J].Arch Dermatol Res,1987,279 Suppl:66-72.
[15]Poochareon V,Berman B,Villa A.Successful treatment of butcher' s warts with imiquimod 5% cream[J].Clin Exp Dermatol,2003,28 Suppl 1:42-44.
[16]Shamanin V,zur Hausen H,Lavergne D,et al.Human Papillomavirus Infections in Nonmelanoma Skin Cancers From Renal Transplant Recipients and Nonimmunosuppressed Patients[J].J Natl Cancer Inst,1996,88(12):802-811.
[17]Fritschi L,Fenwick S,Bulsara M.Mortality and cancer incidence in a cohort of meatworkers[J].Occup Environ Med,2003 Sep,60(9):E4.
[1]Astori G,Lavergne D,Benton C,et al.Human papillomaviruses are commonly found in normal skin of immunocompetent hosts.J Invest Dermatol.1998;110(5):752-5.
[2]Melchers W,de Mare S,Kuitert E,et al.Human papillomavirus and cutaneous warts in meat handlers.J Clin Microbiol.1993Sep;31(9):2547-9.
[3]Keefe M,al-Ghamdi A,Coggon D,et al.Cutaneous warts in butchers.Br J Dermatol.1994 Jan;130(1):9-14.
[4]Keefe M,al-Ghamdi A,Coggon D,et al.Butchers' warts:no evidence for person to person transmission of HPV7.Br J Dermatol.1994Jan;130(1):15-7.
[5]R(u|¨)dlinger R,Bunney MH,Grob R,et al.Warts in fish handlers.Br J Dermatol.1989 Mar;120(3):375-81.
[6]Ritzkowsky A,Weissenborn S,Krieg T,et al.Extensive human papillomavirus type 7-associated orofacial warts in an immunocompetent patient.Acta Derm Venereol.2001 May;81(2):130-3.
[2]Dell G,Gaston K.Human papillomaviruses and their role in cervical cancer[J].Cell Mol Life Sci,2001,58(12-13):1923-1942.
[3]何志秀,刘来奎,陈艳.儿童口腔尖锐湿疣的临床表现及HPV检测[J].华西口腔医学杂志,2002,20(1):27-29.
[4]Kui LL,Xiu HZ,Ning LY.Condyloma acuminatum and human papilloma virus infection in the oral mucosa of children[J].Pediatr Dent,2003,25(2):149-153.
[5]Liu LK,He ZX,Li YN,et al.Infection of human papillomavirus in oral benign epithelial proliferation in children[J].Zbonghua Kou Qiang Vi Xue Za Zhi,2003,38(6):426-428.
[6]蒋明军,王书崎,龚向东.尖锐湿疣皮损中人乳头瘤病毒基因分型研究[J].中华皮肤科杂志,2005,38(5):262-264.
[7]Lu Y,Wang XL,Wu D,et al.Clinical features and epidemiological survey of perianal warts in 72 males[J].Zhonghua Nan Ke Xue,2006,12(10):923-926.
[8]Partridge JM,Koutsky LA.Genital human papillomavirus infection in men[J].Lancet Infect Dis,2006,6(1):21-31.
[9]Anneroth G,Anniko M,Romander H.Oral condyloma accuminatum.A light and electron microscopic study[J].Int J Oral Surg,1982,11(4):260-264.
[10]Piattelli A,Rubini C,Fioroni M,et al.Warty carcinoma of the oral mucosa in an HIV+ patient[J].Oral Oncol,2001,37(8):665-667.
[11]Menzo S,Monachetti A,Trozzi C,et al.Identification of six putative novel human papillomaviruses(HPV) and characterization of candidate HPV type 87[J].J Virol,2001,75(23):11913-11919.
[12]Nishiwaki M,Yamamoto T,Tone S,et al.Genotyping of human papillomaviruses by a novel one-step typing method with multiplex PCR and clinical applications[J].J Clin Microbiol,2008,46(4):1161-1168.
[13]Lin MT,Rohwedder A,Mysliborski J,et al.HPV vulvitis' revisited:frequent and persistent detection of novel epidermodysplasia verruciformis-associated HPV genotypes[J].J Cutan Pathol,2008,35(3):259-272.
[14]余剑敏,沈智君.第二代杂交捕获法与多重荧光PCR在检测高危型人乳头瘤病毒中的应用比较[J].检验医学,2008,23(5):484-487.
[15]李泉,祖洪勇,杨清平,等.实时荧光聚合酶链反应技术与第2代杂交捕获法检测高危亚型人乳头瘤病毒的比较[J].重庆医学,2008,37(24):2802-2804.
[16]唐振华,李柱南,胡伟平.应用实时荧光聚合酶链反应技术与第二代杂交捕获法检测高危亚型人乳头瘤病毒[J].检验医学,2008,23(3):216-219.
[17]姚丽,张伟,王妹妹,等.液相芯片技术原理及应用简介[J].现代肿瘤医学,2008,16(12):2196-2198.
[18]刘思瑶,丁显平,朱一剑,等.液相芯片技术在人乳头瘤病毒检测和分型中的应用[J].四川大学学报自然科学版,2007,44(5):1111-1114.
[1]Astori G,Lavergne D,Benton C,et.al.Human papillomaviruses are commonly found in normal skin of immunocompetent hosts[J].J Invest Dermatol,1998,110(5):752-755.
[2]Berkhout RJ,Tieben LM,Smits HL,et.al.Nested PCR approach for detection and typing of epidermodysplasia verruciformis-associated human papillomavirus types in cutaneous cancers from renal transplant recipients[J].J Clin Microbiol,1995,33(3):690-695.
[3]de Villiers EM,Whitley C,Gunst K.Identification of new papillomavirus types[J].Methods Mol Med,2005,119:1-13.
[4]蒋明军.尖锐湿疣中人乳头瘤病毒感染状况的基因分型研究[D].北京:中国协和医科大学,2004:52-53.
[5]Takahashi S,Tsukamoto T.Condyloma acuminatum[J].Nippon Rinsho,2009,67(1):153-156.
[6]Kondo K.Development of an HPV vaccine-remaining issues and perspective[J].Nippon Rinsho,2009,67(1):62-68.
[7]Narisawa-Saito M,Kiyono T.Mechanisms of high-risk human papillomavirus-induced cervical carcinogenesis[J].Nippon Rinsho,2009,67(1):53-61.
[8]Thierry F.Transcriptional regulation of the papillomavirus oncogenes by cellular and viral transcription factors in cervical carcinoma[J].Virology,2009,20;384(2):375-379
[9]Zekri AR,Bahnassy AA,Seif-Eldin WM,et al.Role of human papilloma virus(HPV) in common and genital warts and its relation to P53expression[J].J Egypt Natl Canc Inst,2006,18(2):117-124.
[10]Oltersdorf T,Campo MS,Favre M,et al.Molecular cloning and characterization of human papillomavirus type 7 DNA[J].Virology,1986,149(2):247-250.
[11]Delius H.,Hofmann B.,Primer-directed sequencing of human papillomavirus types[J].Curr Top Microbiol.Immunol,1994,186:13-31.
[12]de Villiers EM,Neumann C,Oltersdorf T,et al.Butcher's wart virus (HPV 7) infections in non-butchers[J].J Invest Dermatol,1986,87(2):236-238.
[13]Greenspan D,de Villiers EM,Greenspan JS,et al.Unusual HPV types in oral warts in association with HIV infection[J].J Oral Pathol,1988,17(9-10):482-488.
[14]de Villiers EM.Prevalence of HPV 7 papillomas in the oral mucosa and facial skin of patients with human immunodeficiency virus[letter][J].Arch Dermatol,1989,125(11):1590.
[15]R(u|¨)dlinger R,Bunney MH,Grob R,et al.Warts in fish handlers[J].Br J Dermatol,1989,120(3):375-81.
[16]Meffert JJ,Anthony JS.Butcher' s warts:dermatological heritage or testable misinformation?[J].Arch Dermatol,2001,137(3):384-385.
[17]Lei YJ,Gao C,An R,Shi Q,et al.Development of a multiplex PCR method for detecting and typing human papillomaviruses in verrucae vulgaris[J].J Virol Methods,2008,147(1):72-77.
[18]Yan-JunLEI,Chen GAO,Chen WANG,et al.Molecular Epidemiological Study on Prevalence of Human Papillomaviruses in Patients with Common Warts in Beijing[J].Area iomedical and Environmental Sciences,2009,22(1):55-61.
[1]Jablonska S,Obalek S,Favre M,et al.The morphology of butchers' wans as related to papillomavirus types[J].Arch Dermatol Res,1987,279Suppl:66-72.
[2]de Villiers EM,Neumann C,Oltersdorf T,et al.Butcher's wart virus (HPV 7) infections in non-butchers[J].J Invest Dermatol,1986,87(2):236-238.
[3]赵辨.临床皮肤病学[M].北京:人民卫生出版社,2001:313.
[4]Aroni K,Lazaris AC,Ioakim-Liossi A,et al.Histological diagnosis of cutaneous "warty" carcinoma on a pre-existing HPV lesion[J].Acta Derm Venereol,2000,80(4):294-296.
[5]蒋明军,王书崎,龚向东.尖锐湿疣皮损中人乳头瘤病毒基因分型研究[J].中华皮肤科杂志,2005,38(5):262-264.
[6]Lu Y,Wang XL,Wu D,et al.Clinical features and epidemiological survey of perianal warts in 72 males[J].Zhonghua Nan Ke Xue,2006,12(10):923-926.
[7]Partridge JM,Koutsky LA.Genital human papillomavirus infection in men[J].Lancet Infect Dis,2006,6(1):21-31.
[8]Noel JC,Peny MO,Goldschmidt D,et al.Human papillomavirus type 1DNA in verrucous carcinoma of the leg[J].J Am Acad Dermatol,1993,29(6):1036-1038.
[9]Noel JC,Peny MO,Detremmerie O,et al.Demonstration of human papillomavirus type 2 in a verrucous carcinoma of the foot[J].Dermatology,1993,187(1):58-61.
[10]Bragg JW,Ratner D.Human papillomavirus type 2 in a squamous cell carcinoma of the finger[J].Dermatol Surg,2003,29(7):766-768.
[11]Chen AC,McMillan NA,Antonsson A.Human papillomavirus type spectrum in normal skin of individuals with or without a history of frequent sun exposure[J].J Gen Virol,2008,89(Pt 11):2891-2897.
[1]http://www.genscript.com/puc57,html
[2]于永利,麻彤辉.TA克隆及双链DNA:测序:介绍一种快速克隆及分析PCR产物的方法[J].中国免疫学杂志,1994,10(1):5-7.
[3]丁鹏,王家宁,黄永章,等.利用TA克隆载体构建pET15b-SOD1重组质粒[J].郧阳医学院学报,2005,24(2):65-68.
[4]de Villiers EM,Whitley C,Gunst K.Identification of new papillomavirus types[J].Methods Mol Med,2005,119:1-13.
[5]姚丽,张伟,王妹妹,等.液相芯片技术原理及应用简介[J].现代肿瘤医学,2008,16(12):2196-2198.
[6]刘思瑶,丁显平,朱一剑,等.液相芯片技术在人乳头瘤病毒检测和分型中的应用[J].四川大学学报自然科学版,2007,44(5):1111-1114
[7]Astori G,Lavergne D,Benton C,et.al.Human papillomaviruses are commonly found in normal skin of immunocompetent hosts[J].J Invest Dermatol,1998,110(5):752-755.
[8]Berkhout RJ,Tieben LM,Smits HL,et al.Nested PCR approach for detection and typing of epidermodysplasia verruciformis-associated human papillomavirus types in cutaneous cancers from renal transplant recipients[J].J Clin Microbiol,1995,33(3):690-695.
[9]孙海汐,蔡金挺,朱琳楠,等.一种改良的人DNA微量快速检验技术[J].中国实验诊断学,2007,11(6):788-790.
[10]陈辉,刘永波,谢庆瑞,等.有机酚法和Chelex-100法提取不同组织微量DNA 效果比较[J].郑州大学学报医学版,2004,39(6):988-991.
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