人乳头瘤病毒型别及载量与寻常疣临床特征相关性研究

Association analysis between human papillomavirus genotypes and viral load and clinical features of verruca vulgaris

目的 探讨人乳头瘤病毒型别、载量与寻常疣临床特征的相关性.方法 提取48例寻常疣组织DNA;正反向测序通用引物PCR扩增片段,通过blast比对分型,并通过型别特异性引物PCR来验证和补充测序结果;实时荧光定量PCR相对定量ACt法检测HPV病毒载量;HE染色法观察病理变化.结果 35例HPV PCR检测阳性的标本中HPV 7型32例;HPV 57型1例,为四肢末端多发;HPV 2和HPV 7混合感染2例,为头面部、躯干多发病例.单发组与多发组、病程＜6个月组与病程6个月至1年组病毒载量、空泡化细胞数目无明显差异,1年后病毒载量有下降趋势,病程＞2年的皮损,角化过度更明显,空泡化细胞减少.结论 寻常疣的HPV感染型别主要是HPV 7;HPV 7型感染好发于头面部;对于病程1年内的寻常疣,HPV病毒载量及空泡化细胞数目与其数目、病程长短无明显相关性.

Objective To study the association between human papillomavirus （HPV） genotypes and viral load and clinical features of verruca vulgaris.Methods Tissue samples were collected from 48 outpatients with verruca vulgaris,and DNA was extracted from these tissue samples.To determine the genotype of HPV,PCR was performed to amplify the L1 fragment of HPV with universal primers followed by bidirectional sequencing and BLAST.The genotyping results were validated by PCR with type-specific primers.Real-time fluorescence-based quantitative PCR was conducted to measure the viral load of HPV,and hematoxylin and eosin （HE） staining to observe histological changes in these tissue specimens.Results The L1 fragment of HPV was amplified from 35 out of the 48 tissue specimens.Of the 35 L1-positive specimens,32 harbored HPV 7,1 harbored HPV 57,and 2 harbored both HPV 2 and HPV 7.Multiple lesions were observed on extremities in the patient infected with HPV 57,but on the head,face and trunk in the patients coinfected with HPV 2 and HPV 7.There were no significant differences in HPV viral load or vacuolated cell number between patients with single lesions and those with multiple lesions,or between patients with a clinical course of 〈 6 months and those with a clinical course of 6-12 months.However,HPV viral load tended to decrease one year after the onset,and there was pronounced hyperkeratosis and less vacuolated cells in lesions of long duration （more than 2 years） compared with those of short duration （less than 2 years）.Conclusions HPV 7 appears to be the most common HPV genotype associated with verruca vulgaris,and HPV 7 infection usually occurs on the head and face.For verruca vulgaris of less than 1 year,neither HPV viral load nor vacuolated cell number is associated with the count or clinical course of warts.