【摘要】：腸道病毒71型(Enterovirus71，EV71)是小核糖核酸病毒科腸道病毒屬，單鏈的小RNA腸道病毒，其感染性強(qiáng)且致病率高，對中樞神經(jīng)系統(tǒng)的侵害尤其嚴(yán)重。近年來，EV71引起社會各界的關(guān)注是因?yàn)榘l(fā)現(xiàn)該病毒是兒童手足口病(HFMD)的主要病原體之一，特別是導(dǎo)致6歲以下兒童重癥手足口�。℉FMD）的發(fā)生。據(jù)統(tǒng)計，目前手足口病90%以上的重癥病例是由EV71引起，患兒除了有HFMD的典型癥狀外，還有高達(dá)11．1％和0．6％的感染者患有無菌性腦膜炎、腦炎的中樞神經(jīng)系統(tǒng)疾病，且病程進(jìn)展迅速，導(dǎo)致近年來死亡率都高于25/1000。EV71的爆發(fā)流行嚴(yán)重危害我國兒童的身體健康，給廣大民眾帶來極大的心理恐慌�，F(xiàn)階段國內(nèi)外尚未研制出針對EV71的疫苗、特異性治療藥物，，對于控制HFMD的發(fā)生主要依靠對低幼齡兒童進(jìn)行宣傳教育，建立良好的衛(wèi)生習(xí)慣，因此研究安全，高效，經(jīng)濟(jì)的疫苗迫在眉睫。 本研究以EV71表位肽疫苗研究為著手點(diǎn)，通過比較29個國內(nèi)流行株（來自福建，安徽阜陽，江蘇，浙江等省份）C4型基因序列，以及28個NCBI基因庫中收錄的B型EV71異型毒株的基因序列，發(fā)現(xiàn)5個表位在不同血清型的病毒株中能保持很高的同源性。以日本腦炎的D蛋白為載體蛋白與上述5個表位以T-T-B和T-B-B的方式串聯(lián)，一共設(shè)計構(gòu)建了8種表位肽組合，并在原核表達(dá)系統(tǒng)中進(jìn)行了高效表達(dá)。 以原核表達(dá)的重組蛋白為抗原進(jìn)行的免疫性試驗(yàn)，結(jié)果顯示，上述8種組合均能在小鼠血清中的產(chǎn)生抗體；其中C，E，F(xiàn)，H四種組合能產(chǎn)生較高水平的抗體，達(dá)到本研究的設(shè)計目的，期望可為EV71新型疫苗設(shè)計提供理論基礎(chǔ)，研制出有應(yīng)用價值的EV71表位肽疫苗。
[Abstract]:Enterovirus 71 (Enterovirus71,EV71) is a small RNA enterovirus belonging to small ribonucleic acid virus family. It is highly infectious and has a high pathogenicity, especially to the central nervous system (CNS). In recent years, EV71 has attracted the attention of all walks of life because it has been found to be one of the major pathogens of HFMD in children, especially (HFMD) in children under 6 years of age. According to statistics, at present, more than 90% of severe cases of hand, foot and mouth disease are caused by EV71. In addition to the typical symptoms of HFMD, 11. 1% and 0. 6% of the infected patients also suffer from aseptic meningitis and encephalitis central nervous system diseases. The rapid progress of the course of disease has led to the death rate higher than the outbreak of 25/1000.EV71 in recent years the outbreak of epidemic serious harm to the health of children in our country and bring great psychological panic to the general public. At present, there has not been developed a vaccine for EV71 and a specific therapeutic drug at home and abroad. In order to control the occurrence of HFMD, it mainly depends on the propaganda and education of young children and the establishment of good hygiene habits. Therefore, the research is safe and efficient. Economic vaccines are imminent. In this study, EV71 epitope peptide vaccine was used as the starting point to compare the C4 gene sequences of 29 prevalent strains (from Fujian, Fuyang, Jiangsu and Zhejiang provinces) in China. The gene sequences of B type EV71 heterotypic strains collected from 28 NCBI gene banks showed that 5 epitopes could maintain high homology in different serotype strains. Eight epitope peptides were designed and expressed in prokaryotic expression system by using Japanese encephalitis D protein as carrier protein in tandem with the above five epitopes by T-B and T-B-B. The results of immunological tests using prokaryotic expressed recombinant protein as antigen showed that all the above eight combinations could produce antibodies in mouse serum, among which four combinations of CfG and FG could produce a higher level of antibodies, so as to achieve the purpose of the design of this study. It is expected to provide a theoretical basis for the design of new EV71 vaccine and to develop EV71 epitope peptide vaccine.
【學(xué)位授予單位】：河南師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.1

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