尖锐湿疣样本中HPV病毒的分子检测  

Molecular Detection of Specific HPV Types in Condylomata Acuminata

作　　者：吴晓静[1] 任建功[1] 刘永江 刘军连[3] 汪建军[4] 谭萍萍[1] 冉多良[5] 马润林[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所,北京100101 [2]威海迪沙麦特生物制品有限责任公司,威海264206 [3]中国人民解放军第306医院,北京100101 [4]北京安贞医院,北京100029 [5]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《遗传》2005年第5期699-704,共6页Hereditas（Beijing)

基　　金：威海迪沙麦特生物制品有限责任公司HPV专项(编号:20040303)~~

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黏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%。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。To detect HPV in genital warts （Condylomata acuminata, OA） 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-POR was conducted in duplication for each OA sample from male and female patients. DNA of TA-cloned HPV was used as positive control, and deionized H20 was used as negative control. A total of 22 clinical samples, 13 from males and 9 from females, was collected from patients diagnosed with OA 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 HPV11 was 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.

关 键 词：尖锐湿疣 人乳头瘤病毒 临床诊断 

分 类 号：R752.53[医药卫生—皮肤病学与性病学]