HPV L1共同保守序列多肽对多价性HPV阳性临床标本的检测  

The peptide containing the conserved sequence of HPV L1 was used in the detection of polyvalent HPV-positive clinical specimens

作　　者：姜波玲[1] 肖长义[1] 叶红[2] 

机构地区：[1]三峡大学医学院免疫教研室,宜昌443002 [2]三峡大学第一临床医学院,湖北省宜昌市中心医院妇产科,宜昌443003

出　　处：《肿瘤》2008年第10期864-868,共5页Tumor

基　　金：国家自然科学基金资助项目(编号:30770104);湖北省卫生厅科研基金资助项目(编号:JX3B32);湖北省教育厅自然科学研究资助重大项目(编号:Z200713002)

摘　　要：目的:证实HPV16 L1 C-末端448-477氨基酸序列多肽所诱导的免疫抗血清能够与多种型别HPV L1发生反应。方法:人工合成HPV L1共同保守序列短肽,用该合成短肽免疫动物获得抗短肽多克隆抗血清。用PCR方法对宫颈癌、尖锐湿疣等临床标本进行HPV型别鉴定,然后用ELISA、Western印迹、免疫组织化学3种方法检验已获得的抗HPV短肽血清抗体能否与多型别HPV阳性临床标本进行反应。结果:在ELISA实验中,HPV6,11,16,18型及其他型阳性的临床标本的抗原抗体反应均为阳性;Western印迹实验中,HPV各型别阳性的临床标本均在56×103处出现显色条带,而阴性标本没有;免疫组织化学结果发现,不同病变性质的组织反应各有特征,阳性反应仅出现于上皮之内,呈区域性,有明显界限。结论:证实了本研究寻找到的HPV L1共同保守序列确实含有HPV L1 B-细胞共同表位,其免疫形成的抗血清能与多型别HPV L1发生反应。这一发现对今后研制广谱HPV L1疫苗或广谱检测试剂盒具有重要的启示作用。Objective： To confirm that the antiserum induced by short peptide in the region of AA 448-477 at HPV16 L1 C-terminal can respond with many types of HPV L1. Methods： This sutdy synthesized a short peptide containing the conserved sequence of HPV L1 and immunized animal with the synthetic peptide to obtain the polyclonal antiserum. The type of HPV in clinical specimens, such as cervical cancer and condyloma acuminatum tissues, was identified by PCR. ELISA, Western blotting, and immunohistochemis- try were used to determine whether the anti-serum of the short peptide could react with different types of HPV-positive specimens. Results： ELISA assay showed that the reaction between antigen and antibody was positive in HPV6, 11, 16, and 18 and other HPV- positive clinical specimens. Western blotting revealed that there was a strip at 56 kDa in all types of HPV-positive clinical specimens, but there was no strip in HPV-negative clinical specimens. Immunohistochemistry analysis demonstrated that the histological reaction had varying features in different clinical specimens. The positive-reaction appeared only in the epithelium tissues, which was regional and had apparent borders. Conclusion： This study confirms that the common conserved sequence really has HPV L1 common epitope. Its anti-serum can respond with many types of HPV L1. These findings provide an important basis for further researching a broad-spectrum HPV L1 vaccine or publicizing broad-spectrum HPV detection kits.

关 键 词：宫颈肿瘤 乳头瘤病毒 人 免疫血清 诊断试验 常规 表位 

分 类 号：R737[医药卫生—肿瘤]