利用重组HPV16L1蛋白检测鲍温病和鲍温样丘疹病血清中的抗体  

Recombinant Human Papillomavirus Type 16 Capsid Protein L1 on Detecting Antibodies in Patients with Bowenoid Papulosis and Bowen's Disease

作　　者：唐旭[1] 沈宏[1] 刘继峰[1] 许爱娥[1] 

机构地区：[1]杭州市第三人民医院,杭州310009

出　　处：《中国中西医结合皮肤性病学杂志》2010年第2期83-86,共4页Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine

基　　金：杭州市科技局医学重点专科专病项目(2004433Q05)

摘　　要：目的用原核表达系统获得人乳头瘤病毒16型(HPV16)主要衣壳蛋白(L1),检测鲍温病和鲍温样丘疹病血清中的HPV16L1IgG抗体。方法PCR方法检测鲍温病和鲍温样丘疹病局部皮损中的HPVDNA,限制性片段长度多肽性方法进行HPV分型;用原核表达载体pET32a表达HPV16L1蛋白,亲和层析法对表达产物进行纯化,获得了HPV16L1蛋白抗原;ELISA法检测鲍温病和鲍温样丘疹病血清中的抗体。结果用原核表达系统和镍柱亲和层析获得了较纯的HPV16L1蛋白。鲍温样丘疹病血清中抗体的检测率为(20/50)40%,鲍温病血清中抗体的检测率为(8/30)26.7%,正常献血员组检测率为(2/50)4%。这两组分别与正常献血员组比较有统计学差异(P<0.05)。两组HPV16DNAPCR检测法与血清HPV16L1抗体检测法有较好的一致性(P<0.05)。结论利用原核表达系统获得纯化HPV16型L1蛋白,可用于鲍温样丘疹病和鲍温病血清中HPV16抗体的检出,为其临床上血清学诊断提供参考,进一步开发HPV相关皮肤恶性肿瘤高危人群的诊断试剂盒提供前沿基础研究。Objective To produce human papillomavirus（HPV）type 16 major capsid protein （L1） by prokaryotic expression vector, and to screen antibodies in the sera of Bowenoid papulosis and Bowen＇s disease patients with the purified HPV16 L1 protein as antigen. Methods PCR was used to detect the HPV DNA in the skin lesion of Bowenoid papulosis and Bowen＇s disease patients. The HPV genotype was determined by restriction fragment length polymorphisms. HPV16 L1 protein was produced by prokaryotic expression vector pET32a and purified with affinity chromatography. Using the purified HPV16 L1 protein as the antigen, we analysed by enzyme-linked immunosorbent assay（ELISA）the sera of Bowenoid papulosis and Bowen＇s disease for the presence of IgG antibodies. Results Using HPV16 L1 protein as antigen, IgG antibodies were detected in sera from 20 of 50（40.0%）. Bowenoid papulosis patients and 8 of 30 （26.7%） Bowen＇s disease both had a statistically significant difference compared with the control group （4%）. A consistency was also found between the results of the PCR method to detect HPV16 DNA, and the ELISA method to detect HPV16 IgG antidodies. Conclusion After being produced by prokaryotic expression system and purified with affinity chromatography, HPV16 L1 protein can be used as the antigen to detect the IgG antibodies in sera from Bowenoid papulosis and Bowen＇s disease patients. The results bring valuable reference for the serological diagnosis of HPV16 infection. Furthermore, diagnostic ELISA kits might be developed to detect the risk population of HPV-associated skin malignancies.

关 键 词：鲍温样丘疹病 鲍温病 人乳头瘤病毒16型 主要衣壳蛋白 酶联免疫法 

分 类 号：R730.26[医药卫生—肿瘤]