外阴白斑组织中人乳头瘤病毒的快速检测与基因分型

Rapid detection and genotyping of human papillomavirus in patients with vulvar leukoplakia

目的 为探讨人乳头瘤病毒 (HPV)与外阴白斑发生的关系 ,应用通用引物聚合酶链式反应和反向点探针杂交方法对 5 8例外阴白斑进行 HPV6 ,1 1 ,1 6 ,1 8,3 1 ,3 3等型别进行快速检测及基因分型 .方法 将上述 6个病毒型别的特异性寡核苷酸探针依次固定在尼龙膜上 ,再与经通用引物 PCR扩增的 DNA靶序列杂交 ,即可在同一张膜上分辨出 6型 HPVDNA的任一型或几型 .结果 在我们检测的 5 8例外阴白斑患者中 ,总阳性检出率为 3 7.9% (2 2 /5 8) .其中 ,HPV6型 7例 (3 1 .8% )、HPV1 1型 5例 (2 2 .7% )、HPV6 /1 1型 (混合型 ) 2例 (9.1 % )、HPV1 6型 1 0例 (45 .5 % )、HPV1 8型 1例(4.5 % )、HPV3 1型 0例、HPV3 3型 1例 (4.5 % ) .经统计学分析 ,外阴白斑的发生与患者感染 HPV有显著相关性 (P<0 .0 1 ) .结论 良性型 HPV(HPV6 ,1 1 ,6 /1 1 )与恶性型 HPV(HPV1 6 ,1 8,3 3 )

AIM To establish the correlation between the infection of human papillomavirus and occurrence of vulvar leukoplakia. METHODS A reverse hybridization blot probe assay was used.Six sequence specific oligonucleotides of HPVDNA (types 6, 11, 16, 18, 31 and 33) were used as specific probes and were fixed separately on a strip of nylon membrane, which could be used to hybridize with the PCR amplified target DNA sequence, so that we could detect and identify any one of the 6 types of HPV on the same strip. Paraffin sections of 58 patients with vulvar leukoplakia were tested for the presence of HPV by this assay and a general primer based PCR. RESULTS 22 out of 58 (37.9%) specimens tested were positive for HPV DNA by PCR, of which 7 were positive for HPV6 (31.8%), 5 for HPV11 (22.7%), 2 for HPV6/11 (mixed type) (9.1%), 10 for HPV16 (45.5%), 1 for HPV18 (4.5%), 0 for HPV31, 1 for HPV33 (4.5%). Statistical analysis revealed that there was marked correlation between the pathogeny of vulvar leukoplakia and HPVs ( P <0.01). CONCLUSION Infection of human papillomavirus may play a role in the occurrence of vulvar leukoplakia. We have identified both the benign and the malignant type HPVs in the vulvar leukoplakias.