人細小病毒B19VP2蛋白的優(yōu)勢表達與抗原活性檢測
發(fā)布時間：2019-03-20 18:44
【摘要】：研究目的： 人細小病毒B19(Human parvovirus B19)與人類多種疾病發(fā)生有密切關系，可引起特發(fā)性血小板減少性紫癜、急性再生障礙性貧血、幼年類風濕關節(jié)炎、過敏性紫癜等多種疾病，并可造成胎兒水腫、死胎、流產和先天畸形，，給患者生命和健康造成嚴重影響。B19病毒基因組主要編碼兩種結構蛋白VP1(85KDa)和VP2(58KDa)以及一種非結構蛋白NS(77KDa)。VP1和VP2均具有抗原性，能刺激機體產生相應的IgM和IgG抗體，可用于B19病毒感染的診斷。免疫功能正常兒童感染B19病毒后可產生特異性抗體，B19-VP2-IgM陽性提示近期感染或急性感染，發(fā)病后3天即可檢出，2-3周達高峰，持續(xù)2-3個月，適用于B19病毒感染的早期診斷。但由于B19病毒抗原來源困難，國內相關的血清學檢測試劑很少，臨床診斷大都采用昂貴的國外進口ELISA試劑盒。 分子生物學技術的發(fā)展使得許多病毒衣殼蛋白能夠在體外進行基因重組表達，這也為解決B19病毒抗原來源難的問題開辟了一條新的途徑。本研究通過對B19病毒VP2序列的生物信息學分析，選定VP2片段中4354-4662bp(309bp，351-454位氨基酸)表面易得性較高的片段作為研究對象，利用分子生物學方法獲得VP2基因片段及重組的VP2蛋白，再利用ELISA方法對該重組蛋白的生物學功能進行初步分析。 研究內容與結果：
[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:
【學位授予單位】：中國人民解放軍軍事醫(yī)學科學院
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R373
本文編號：2444510
[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:
【學位授予單位】：中國人民解放軍軍事醫(yī)學科學院
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R373
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