本文選題：黃熱病病毒 + 黃熱疫苗��； 參考：《北京生物制品研究所》2010年碩士論文

【摘要】： 黃熱疫苗(YFV)自1936年開(kāi)始在黃熱病流行地區(qū)用于預(yù)防黃熱病的感染,已經(jīng)在世界范圍內(nèi)安全使用了60多年,有效控制了黃熱病的大規(guī)模流行,是目前最安全、有效的蟲(chóng)媒病毒疫苗之一。但是至今尚未完全控制非洲和南美洲地區(qū)的黃熱病爆發(fā),除了疫苗的推廣使用未能覆蓋所有易感人群外,越來(lái)越多的黃熱疫苗相關(guān)不良反應(yīng)也引起了世界衛(wèi)生組織和科學(xué)家的高度重視。 黃熱疫苗病毒在雞胚卵黃囊中培養(yǎng),因其特有的培養(yǎng)基質(zhì)和病毒特性,在后續(xù)純化工藝中,很難在保持有效免疫原性的同時(shí)獲得良好的純度。本實(shí)驗(yàn)通過(guò)改變其培養(yǎng)基質(zhì),以期在不降低免疫原性的同時(shí),減少疫苗中雞胚雜質(zhì)殘留。將17D黃熱疫苗工作毒種在雞胚成纖維細(xì)胞中連續(xù)傳代,通過(guò)對(duì)傳代病毒的生物學(xué)相關(guān)特性和基因遺傳穩(wěn)定性研究,探討將傳統(tǒng)用于黃熱疫苗生產(chǎn)的全雞胚改換為雞胚成纖維細(xì)胞后,是否會(huì)對(duì)疫苗病毒的基因組和病毒的生物學(xué)特性產(chǎn)生影響,從而為使用雞胚成纖維細(xì)胞作為黃熱疫苗生產(chǎn)基質(zhì)的可能性提供依據(jù)。 本實(shí)驗(yàn)選擇黃熱疫苗國(guó)際標(biāo)準(zhǔn)品99/616-CE(天壇生物編號(hào))和我國(guó)黃熱疫苗工作毒種2008(54)01(54代)5#(天壇生物編號(hào))作為實(shí)驗(yàn)用毒種,99/616為世界衛(wèi)生組織參考品。兩者都是17D疫苗株,均源于17D204株,并經(jīng)特異性抗血清鑒別試驗(yàn)確認(rèn)毒種。兩株毒種分別在雞胚成纖維細(xì)胞中進(jìn)行適應(yīng)性傳代,各自盲傳10代,記作“99/616-傳代病毒代次”和“YFVBJ-傳代病毒代次”。并分別對(duì)兩株病毒的傳代病毒的生物學(xué)相關(guān)特性和基因遺傳穩(wěn)定性進(jìn)行了研究,內(nèi)容包括傳代病毒病毒滴度檢測(cè)、LD50檢測(cè)、小鼠平均生存時(shí)間檢測(cè)、蝕斑大小檢測(cè)、免疫原性檢測(cè)、雜蛋白含量以及非結(jié)構(gòu)基因NS1序列測(cè)定。 實(shí)驗(yàn)結(jié)果的數(shù)據(jù)分析如下: 一.兩株毒種的傳代病毒滴度都存在一個(gè)下降趨勢(shì),但滴度均在5.0lgPFU/ml以上,各代次之間差異不顯著。 二.兩株毒種的傳代病毒經(jīng)LD50檢測(cè)都存在一個(gè)下降趨勢(shì),但除99/616-1外滴度都在5.0lgLD50以上,各代次之間差異不顯著。 三.小鼠平均生存時(shí)間、免疫原性等研究表明,傳代病毒各代次之間未見(jiàn)顯著性統(tǒng)計(jì)學(xué)差異。 四.兩株毒種的傳代病毒在蝕斑大小試驗(yàn)中顯示蝕斑大小存在變小的趨勢(shì),部分代次之間統(tǒng)計(jì)學(xué)差異顯著。 五.YFVBJ各傳代病毒非結(jié)構(gòu)基因NS1序列分析表明堿基序列未發(fā)生改變。這些結(jié)果提示雖然改變了培養(yǎng)基質(zhì),但我國(guó)的黃熱疫苗工作毒種無(wú)論在生物學(xué)相關(guān)特性還是基因序列遺傳穩(wěn)定性上都表現(xiàn)出良好的穩(wěn)定性。 綜上所述:本研究完成了黃熱疫苗工作毒種和99/616黃熱國(guó)際標(biāo)準(zhǔn)品在雞胚成纖維細(xì)胞中的傳代,并初步研究了培養(yǎng)基質(zhì)改變對(duì)黃熱病病毒的生物學(xué)相關(guān)特性以及基因遺傳穩(wěn)定性的影響,除了傳代病毒的蝕斑大小有明顯的差異之外,傳代病毒表現(xiàn)出了良好的穩(wěn)定性。這些工作為今后更深入的研究以雞胚成纖維細(xì)胞為基質(zhì)培養(yǎng)17D黃熱病疫苗提供了基礎(chǔ)數(shù)據(jù)。
[Abstract]:Yellow fever vaccine ( YFV ) has been used for the prevention of yellow fever in the epidemic area of yellow fever since 1936 . It has been safely used in the world for more than 60 years . It is one of the most secure and effective vaccine for the disease of yellow fever . However , it has not fully controlled the outbreak of yellow fever in Africa and South America . Besides , it has not completely controlled the outbreak of yellow fever in Africa and South America . In addition to the popularization and use of the vaccine , more and more adverse reactions related to yellow fever vaccine have also attracted much attention from the World Health Organization and scientists .



in ord to reduce immunogenicity , it is difficult to obtain good purity while maintaining effective immunogenicity .



In this experiment , the international standard of yellow fever vaccine 99 / 616 - CE ( Tiantan Biological No . ) and China Yellow Fever Vaccine Working Poison 2008 ( 54 ) 01 ( 54 generation ) 5 # ( Tiantan Biological No . ) were selected as the reference products for the experiment . Both of them were derived from 17D204 strains .



The data analysis of the experimental results is as follows :



One . There was a downward trend in the titer of the two strains of virus , but the titer was above 5.0lgPFL / ml , and there was no significant difference between successive generations .



2 . There was a downward trend in the LD50 test of the two strains of the virus , but the titer was more than 5.0g LD50 except for 99 / 616 - 1 , and there was no significant difference between the generations .



3 . The average survival time and immunogenicity of mice showed that there was no significant statistical difference between the generations of the passaging virus .



4 . There was a tendency for the size of plaque to be smaller in the test of plaque size . There was a significant difference between the partial generations .



5 . The analysis of the non - structural gene of the non - structural gene of YFVBJ showed that the nucleotide sequence had not changed . These results suggested that although the culture substrate was changed , the yellow fever vaccine work in China showed good stability both in the biology - related characteristic and the genetic stability of the gene sequence .



In conclusion , this study has completed the passage of yellow fever vaccine work toxin and 99 / 616 yellow fever international standard in chicken embryo fibroblasts , and preliminarily studied the effect of the change of culture matrix on the biological characteristics and genetic stability of yellow fever virus .
【學(xué)位授予單位】：北京生物制品研究所
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 李淑云,黃浩,劉瑜tD,星海,高靜,唐巧英;應(yīng)用蝕斑法滴定17D黃熱疫苗效力[J];中國(guó)生物制品學(xué)雜志;2000年03期

2 李淑云;付琨;黃浩;徐程林;付永琪;;雞胚細(xì)胞制備17D黃熱疫苗[J];中國(guó)生物制品學(xué)雜志;2007年11期

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本文編號(hào)：1975773