遗传 ›› 2005, Vol. 27 ›› Issue (5): 699-704.

• 研究报告 • 上一篇    下一篇

尖锐湿疣样本中HPV病毒的分子检测

吴晓静1;2;任建功1;刘永江3;刘军连4;汪建军5;谭萍萍1;冉多良2;马润林1   

  1. 1.中国科学院遗传与发育生物学研究所,北京100101;2.新疆农业大学动物医学学院,新疆830052;
  • 收稿日期:2005-03-24 修回日期:2005-05-10 出版日期:2005-10-10 发布日期:2005-10-10
  • 通讯作者: 马润林

Molecular Detection of Specific HPV Types in Condylomata Acuminata

LIU Jun-Lian4, WANG Jian-Jun5, TAN Ping-Ping1, RAN Duo-Liang2, MA Run-Lin1   

  1. WU Xiao-Jing1,2, REN Jian-Gong1, LIU Yong-Jiang3, 1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; 2. College of veterinary medicine,Xinjiang Agricultural University,Xinjiang 830052, China; 3.Weihai Disha-Matt Biotechnology Company, Shandong 264205, China; 4. 306th Hospital of People’s Liberation Army, Beijing 100101, China; 5.Department of OBGYN, Anzhen Hospital, Beijing 100029, China
  • Received:2005-03-24 Revised:2005-05-10 Online:2005-10-10 Published:2005-10-10
  • Contact: MA Run-Lin

摘要: 调查男性和女性尖锐湿疣样本中人乳头瘤病毒(HPV)的检出率及病毒类型,为研发相关防治疫苗提供依据,以HPV 外壳蛋白DNA序列为模板设计特异引物,SSP-PCR扩增检测样本中HPV的感染率和病毒类型。收集了北京及邯郸市医院门诊尖锐湿疣样本22例,其中男性13例,女性9例。检测发现所有样本中存在着高浓度的HPV病毒DNA。男性样本中有5例感染HPV6型,6例感染HPV11型,2例为HPV6+HPV11混合感染。女性样本中有3例感染HPV6型,2例感染HPV11型,4例为 HPV6+HPV11混合感染。被诊断为宫颈湿疣的4位女性还在其含宫颈粘膜脱落细胞的样本中检出了HPV16、HPV18、HPV33、HPV35、HPV45、HPV54、HPV56或HPV58等高危险型病毒类型。所有检测到的HPV病毒DNA片段均TA克隆并将测定的DNA序列存入了国际基因数据库GenBank(DQ003066-DQ003079)。调查没有在单纯的男女尖锐湿疣组织块中检测到除HPV6和HPV11以外的其他HPV类型。该研究建立了灵敏可靠的HPV分子检测及分型方法,尖锐湿疣中HPV的检出率达100%。 本研究初步结果显示导致男女尖锐湿疣的HPV病毒类型没有显著差异,主要为HPV6及HPV11型。

关键词: 人乳头瘤病毒, 尖锐湿疣, 临床诊断

Abstract: To detect HPV in genital warts (Condylomata acuminata, CA) for infection rate and association of specific HPV types between males and females, and to provide support for the development of HPV vaccines, we designed HPV type-specific oligonucleotide primers to amplify DNA fragments encoding L1 viral capsule protein. SSP-PCR was conducted in duplication for each CA sample from male and female patients. DNA of TA-cloned HPV was used as positive control, and deionized H2O was used as negative control. A total of 22 clinical samples, 13 from males and 9 from females, was collected from patients diagnosed with CA at hospitals in Beijing and Handan. HPV viral DNA was amplified in all 22 samples analyzed, with 100% detection rate. TA-cloning and sequencing of the PCR products confirmed correct amplification of HPV type-specific fragments. Of the 13 samples from males, 5 were infected with HPV6, 6 with HPV11, and 2 with HPV6 + HPV11. Of the 9 samples from females, 3 were infected with HPV6, 2 with HPV11, and 4 with both HPV6 and HPV11 infection. In addition, high-risk types HPV16, HPV18, HPV33, HPV35, HPV45, HPV54, HPV56 and HPV58 were also detected in 4 female samples that were mixed with cervical cell debris during sample collection. However, no HPV types other than HPV6 and HPV11was detected in all CA-only samples in this study. We have established a sensitive and reliable laboratory procedure for HPV detection and classification. Using the method, we reached 100% detection rate of HPV in the CA samples. Our results confirm that HPV6 and HPV11 are primarily responsible for CA, and there is no specific association of HPV types between warts in males and females.

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