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  本文選題：人博卡病毒　切入點：桿狀病毒表達載體系統(tǒng)　出處：《武漢生物制品研究所》2010年碩士論文

【摘要】： 2005年，瑞典學者首次報道在呼吸道疾病患者鼻咽分泌物中發(fā)現(xiàn)人博卡病毒1，隨后，人博卡病毒2和人博卡病毒3又分別在急性遲緩型麻痹患兒和腹瀉患兒的糞便中被相繼發(fā)現(xiàn)。人博卡病毒是一類無包膜的單鏈線狀DNA病毒，暫時劃歸為細小病毒科、細小病毒亞科、博卡病毒屬。 由于缺乏有效的體外細胞培養(yǎng)體系和動物模型，人博卡病毒與人類呼吸道或胃腸道疾病的臨床關(guān)聯(lián)尚且未知。因此，使用基因工程方法大量制備人博卡病毒抗原，建立相應的血清學檢測方法，將有助于解開這些疑問。 本研究使用昆蟲細胞和桿狀病毒表達載體系統(tǒng)表達人博卡病毒1、2和3衣殼蛋白VP2基因，重組衣殼蛋白VP2部可以自行組裝成病毒樣顆粒，使用純化的人博卡病毒樣顆粒建立酶聯(lián)免疫吸附試驗，檢測200份西安地區(qū)健康嬰兒、176份北京地區(qū)健康人血清中人博卡病毒1、2和3的IgG和IgM水平，以及373份日本福岡地區(qū)健康人血清中人博卡病毒1的IgG水平。 電泳及透射電鏡結(jié)果顯示，大量分子量約為60．5kD的蛋白釋放入培養(yǎng)上清中，，重組蛋白可以自行組裝成不含核酸的空心病毒樣顆粒，這些病毒樣顆粒平均直徑為22nm，顆粒密度約為1．31g／ml。人博卡病毒血清流行病學結(jié)果顯示，嬰兒中人博卡病毒1、2和3的IgG陽性率分別為20．0％、14．0％和13．5％；而10歲以上人群中人博卡病毒1、2和3的IgG陽性率升高至75．6％、44．3％和42．0％；但是日本福岡地區(qū)人博卡病毒1的IgG陽性率超過80％。與IgG流行趨勢相反，嬰兒中人博卡病毒的IgM陽性率明顯高于成年人群。 昆蟲細胞和桿狀病毒表達載體系統(tǒng)可以有效表達人博卡病毒1、2和3衣殼蛋白VP2，并自行組裝成病毒樣顆粒。人博卡病毒在普通人群中廣泛流行，成年人群中人博卡病毒的IgG的陽性率很高。
[Abstract]:In 2005, Swedish scholars first reported the discovery of human Boca virus 1 in nasopharyngeal secretions of patients with respiratory diseases. Subsequently, human Boca virus 2 and human Boca virus 3 were found in feces of children with acute delayed paralysis and diarrhea, respectively.Human Boca virus is a class of uncoated single strand linear DNA virus, which is classified as parvovirus family, parvovirus subfamily and Boca virus genus.Due to the lack of effective cell culture system in vitro and animal models, the clinical association between human Boca virus and human respiratory or gastrointestinal diseases is unknown.Therefore, it is helpful to solve these questions by using genetic engineering method to prepare human Boca virus antigen in large quantities and to establish corresponding serological detection method.In this study, the insect cell and baculovirus expression vector system was used to express the VP2 gene of human Boca virus 1 / 2 and 3 capsid protein. The recombinant capsid protein VP2 could be self-assembled into virus-like particles.Enzyme linked immunosorbent assay (Elisa) was established using purified human Boca virus-like particles to detect the levels of IgG and IgM in sera of 200 healthy infants from Xi'an area and 176 healthy people from Beijing area.The IgG level of human Boca virus 1 in 373 healthy people in Fukuoka, Japan.The results of electrophoretic and transmission electron microscopy showed that a large number of proteins with molecular weight of about 60.5kD were released into the supernatant, and the recombinant protein could be self-assembled into hollow virus-like particles without nucleic acid. The average diameter of these virus-like particles was 22 nm and the particle density was about 1.31 g / ml.The results of seroepidemiology of human Boca virus show thatThe positive rates of IgG in infants were 20.0% and 13.50.The IgG positive rates of human Boca virus 1and 3 increased to 75.643% and 42.0%, respectively, but the IgG positive rate of Boca virus 1 was more than 80% in Fukuoka area, Japan.Contrary to the epidemic trend of IgG, the positive rate of IgM in infants was significantly higher than that in adults.Insect cells and baculovirus expression vector system can effectively express human Boca virus 1 / 2 and 3 capsid protein VP2and self-assemble virus-like particles.Human Boca virus is prevalent in the general population, and the positive rate of human Boca virus IgG is very high in adult population.
【學位授予單位】：武漢生物制品研究所
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R373

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相關(guān)期刊論文 前1條

1 瞿小旺;漆正宇;段招軍;劉勁松;劉巧突;劉文培;黃燦平;謝志萍;彭夫望;高寒春;鄭麗舒;侯云德;;兒童急性呼吸道博卡病毒感染[J];病毒學報;2006年02期

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