【作者】 张世杰；

【导师】 赵国强；

【作者基本信息】 郑州大学 ， 免疫学， 2006， 硕士

【摘要】 背景与目的： 人微小病毒B19(Human Parvovirus B19)是微小病毒科微小病毒属中目前已知的唯一使人类致病的病毒，也是动物病毒中体积最小结构最简单的DNA病毒。主要经呼吸道和血液途径传播，可引起社区局部流行。自1990年首次证实我国存在该病毒后，陆续报告B19病毒感染是我国孕妇非免疫性流产、儿童急性再障危象、急性血小板减少性紫癜等疾病的重要病因之一。现己证实该病毒同多种儿童及成人疾病有关。同时，我国妇女和儿童血清中保护性抗体水平低，易发生该病毒感染。尤其免疫功能低下或缺陷患者可发生持续感染和慢性贫血，危及生命。且随着检测手段的改进及研究的深入，其疾病谱还在不断扩大。B19病毒基因组长约5.6kb，编码两个结构蛋白(VP1和VP2)及一个主要的非结构蛋白(NS1)。B19病毒衣壳蛋白主要由VP2蛋白组成，VP1蛋白不超过衣壳蛋白的10％，其独特区暴露在病毒的表面。B19病毒感染引起的机体免疫反应主要是针对其结构蛋白VP1和VP2，而VP1除在其氨基端有—227个氨基酸的独特区外，其余与VP2相同。乙型肝炎病毒(HBV)的感染在人群中比例很高，且感染后体内能形成的主要保护性抗体为针对表面抗原(HBsAg)的表面抗体(抗-HBs)。本课题的目的是选择B19-VP2抗原基因和HBsAg基因重组，利用pGEX-5X-1原核表达载体表达，从而建立表达B19和HBV复合抗原系统，并检测此复合抗原的抗原性，给HBV和B19病毒重组多价疫苗的研制和复合诊断试剂提供抗原。 方法： 根据GenBank收录的HBV及B19病毒序列，设计PCR引物，并在每条引物前分别加入一定的内切酶位点。利用PCR和分子克隆技术，从含HBV病毒全基因的重更多还原

【Abstract】 Background and Objective:Human parvovirus B19(B19), the only parvovirus known to be pathogenic in human, belongs to the genus Parvovirus in the family Parvoviridae. It is the smallest animal DNA virus, and transmitted mainly by respiratory tract and blood, which can cause local epidemic in certain communities. B19 was reported that it is one of the important pathogens causing acute aplasia anemia, non-immunity hydrops, acute trombocytopenic purpura and other diseases since B19 infection had been confirmed its existence in China in 1990. And now it is certified that B19 is associated with a variety of clinical manifestations in adults and children. Women and children in domestic are sensitive to B19 infection because of the low level of the protective antibodies in their sera. B19 infection in those with low immunity function or immunity deficiency can be persistent and can cause chronic anemia, therefore may threaten their lives. And the spectrum of disease about B19 is also wider because of the development of detection methods and research. The genome of B19 is approximately 5.6kb.The genome encodes a non-structural protein(NSl), and two structural proteins(VPl and VP2) that comprise the B19 capsid, the minor structural protein, VP1, and major structural protein, VP2. B19 capsids are composed mainly of VP2 protein, VPlprotein contributes less than 10% of capsid protein, and its specific region is probably更多还原