论     著

尖锐湿疣组织中HPV型别检测及HPV11L1基因的克隆与测序

庄敏1,谷鸿喜1*,魏兰兰1,刘希君1,李迪1,崔洪波1,孔卫兰2

   

[摘  要]  目的  分析黑龙江地区尖锐湿疣组织感染的HPV型别分布情况,并对HPV11型流行株L1基因进行克隆和测序,检测序列变异情况。

方法  提取30例尖锐湿疣组织标本中的DNA,用HPV L1区通用引物及特异性HPV6b111618型引物进行PCR扩增,分析尖锐湿疣组织中感染的HPV型别分布。对其中4HPV11型阳性标本L1基因,经酶切、连接,分别构建pSP73-HPV11L1重组质粒,并对其L1片段进行测序。

结果  30例标本中,HPV 6b 11型感染数分别为11例,各占36.7%;有2例合并HPV6 b /11/16型的混合感染,占总病例数的6.7%1HPV16型单独感染,未鉴定出HPV18型感染,其他型别5例。pSP73HPV11L1重组质粒测序后,4HPV11L1基因序列完全一致,可认为是黑龙江省流行株。其序列与原始株序列相比,仅发现了两处同义点突变。

结论  黑龙江地区尖锐湿疣组织中HPV DNA检出率为100%,以6b11型为主,偶见高危型别16型感染。克隆测序表明黑龙江省流行株HPV11L1基因高度保守。

[关键词]  尖锐湿疣;人乳头瘤病毒;L1 基因

[中图分类号]  R 752.5      [文献标识码]  A   [文章编号]  1001-7089(2005)09-0-0

 Analysis of HPV Types and Cloning and Sequencing of HPV 11 L1 Gene in Acuminate Condyloma

 ZHUANG Min GU HongxiWEI Lanlan,et al

WT5BX(Department of Microbiology Haerbin Medical University Haerbin 150086China)

 Abstract:  Objective   To analyze the viral types and the incidence of human papillomavirus (HPV) in acuminate condyloma biopsy specimens in Heilongjiang province and sequencing of HPV 11 L1 gene.

 Methods   The tissue DNA was extracted from 30 biopsy specimens of acuminate condylomarespectively.The L1 gene of HPV was amplified with common primers followed by 6b1116 and 18 typespecific primers PCR.Four positive L1 fragments of HPV 11 were identified by cloning into plasmid pSP73 and sequencing.

 Results   HPV 6b and HPV 11 were the two most common typesno differences were found between them36.7%  respectively.Two biopsy specimens were multitypes of HPV 6 b /11/16amount for 6.7%. In comparisononly one of the 30 biopsy specimens was type 16 (3.3%) and no type 18 was found.And five specimens16.6% for other HPV types.Sequencing analysis showed the same homology in four clones of HPV type 11 L1 geneonly two synonymous mutations were found comparing with the original one in GeneBank. It indicated this local type of HPV 11 represented the epidemic strain in Heilongjiang province.

 Conclusion   The incidence of HPV infections in all acuminate condyloma biopsy specimens was 100%.The most common types in acuminate condyloma were type 6b and 11whereas the highrisk type 16 or 18 was very low.High conserved L1 sequence of HPV 11 could represente the epidemic strain in Heilongjiang province.

 Key words   Human PapillomavirusHPV;Acuminate condyloma;L1 gene

 

[基金项目]  黑龙江省攻关项目基金(GB02C111),黑龙江省青年基金(QC03C04)

[作者单位]  1 哈尔滨医科大学微生物学教研室,黑龙江 哈尔滨  150086;
              2
哈尔滨医科大学附属第一医院皮肤科,黑龙江哈尔滨  150086

[作者简介]  庄敏(1972-),女,黑龙江省哈尔滨市人,医学博士,副教授,从事人乳头瘤病毒研究。

  [通讯作者]  谷鸿喜,Email:hxgu2432@163.com

 

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