人细小病毒B19VP2蛋白的优势表达与抗原活性检测

发布时间：2019-03-20 18:44

【摘要】：研究目的：人细小病毒B19(Human parvovirus B19)与人类多种疾病发生有密切关系，可引起特发性血小板减少性紫癜、急性再生障碍性贫血、幼年类风湿关节炎、过敏性紫癜等多种疾病，并可造成胎儿水肿、死胎、流产和先天畸形，，给患者生命和健康造成严重影响。B19病毒基因组主要编码两种结构蛋白VP1(85KDa)和VP2(58KDa)以及一种非结构蛋白NS(77KDa)。VP1和VP2均具有抗原性，能刺激机体产生相应的IgM和IgG抗体，可用于B19病毒感染的诊断。免疫功能正常儿童感染B19病毒后可产生特异性抗体，B19-VP2-IgM阳性提示近期感染或急性感染，发病后3天即可检出，2-3周达高峰，持续2-3个月，适用于B19病毒感染的早期诊断。但由于B19病毒抗原来源困难，国内相关的血清学检测试剂很少，临床诊断大都采用昂贵的国外进口ELISA试剂盒。分子生物学技术的发展使得许多病毒衣壳蛋白能够在体外进行基因重组表达，这也为解决B19病毒抗原来源难的问题开辟了一条新的途径。本研究通过对B19病毒VP2序列的生物信息学分析，选定VP2片段中4354-4662bp(309bp，351-454位氨基酸)表面易得性较高的片段作为研究对象，利用分子生物学方法获得VP2基因片段及重组的VP2蛋白，再利用ELISA方法对该重组蛋白的生物学功能进行初步分析。研究内容与结果：
[Abstract]:Objective: human parvovirus B19 (Human parvovirus B19 may cause idiopathic thrombocytopenic purpura and acute aplastic anemia. Childhood rheumatoid arthritis, Henoch-Schonlein purpura and other diseases, and can cause fetal edema, stillbirth, abortion and congenital malformation, The genome of B19 mainly encodes two structural proteins VP1 (85KDa) and VP2 (58KDa) and a non-structural protein NS (77KDa). VP1 and VP2 have antigenicity. Can stimulate the body to produce the corresponding IgM and IgG antibodies, can be used in the diagnosis of B19 virus infection. Specific antibodies could be produced after infection with B19 virus in children with normal immune function. The positive results of B19-VP2-IgM suggested that short-term infection or acute infection could be detected 3 days after onset, and reached the peak at 2-3 weeks and lasted for 2-3 months. It is suitable for early diagnosis of B19 virus infection. However, due to the difficult source of B19 virus antigen, there are few serological detection reagents in China. Most of the clinical diagnosis uses expensive imported ELISA kit. With the development of molecular biology, many viral capsid proteins can be expressed in vitro, which opens up a new way to solve the problem of B19 virus antigen source. In this study, the bioinformatics analysis of the VP2 sequence of B19 virus showed that the fragment of 4354-4662bp (309bp, 351x454 amino acid) with high surface accessibility was selected as the research object. The fragment of VP2 gene and the recombinant VP2 protein were obtained by molecular biological method. The biological function of the recombinant protein was analyzed by ELISA. Content and results of the study:
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R373

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