本文選題：Vero細胞　切入點：重組流感病毒　出處：《中國協(xié)和醫(yī)科大學》2009年碩士論文

【摘要】： H5N1禽流感在全球各地爆發(fā)以來,到2007年底,WHO已經證實感染禽流感致死的人數(shù)超過200人,致死率高于50%。WHO的專家預測禽流感病毒在人體的逐步適應很可能引起一場全球性的流感大爆發(fā),推薦各國開發(fā)生產H5N1人用流感疫苗。目前H5N1人用流感疫苗主要是用雞胚為培養(yǎng)基質開發(fā)生產,H5N1禽流感爆發(fā)時難于在短期內提供足夠的雞胚生產疫苗,另外,雞胚中可能帶有的外源性因子也影響疫苗的質量。Vero細胞是WHO推薦用于生產疫苗的細胞,能夠利用生物反應器進行大規(guī)模培養(yǎng)。與雞胚來源的疫苗比較,在Vero細胞上得到的流感疫苗可以產生較強的體液免疫反應和細胞免疫反應。如果研究出一種用Vero細胞培養(yǎng)的H5N1禽流感疫苗毒株,將能較好的解決現(xiàn)在禽流感疫苗生產存在的一些問題,對防治禽流感有重大意義。該實驗選用H5N1禽流感疫苗株與本實驗室篩選的H3N2 Vero適應株同時接種雞胚,24小時后取尿囊液,加入H3N2 Vero細胞適應株的抗血清中和后在Vero細胞上傳代,獲得H5N1 Vero細胞高產適應株,利用空斑實驗純化毒株。連續(xù)傳代20代后,應用血凝抑制實驗與基因測序鑒定該毒株,并進一步測定該H5N1 Vero細胞適應株的致病性、免疫原性等生物學特性。獲得的H5N1 Vero細胞適應株可用于H5N1禽流感人用疫苗的開發(fā)。
[Abstract]:Since the outbreak of H5N1 bird flu around the world, by the end of 2007, the WHO had confirmed that more than 200 people had died from bird flu. Experts with a fatality rate higher than 50%.WHO predict that the gradual adaptation of the bird flu virus in humans is likely to lead to a global influenza outbreak. It is recommended that various countries develop and produce H5N1 human influenza vaccine. At present, H5N1 human influenza vaccine is mainly developed with chicken embryos as the culture substrate to produce enough chicken embryos to produce vaccines for the outbreak of H5N1 avian influenza in a short period of time. In addition, it is difficult to provide sufficient chicken embryos to produce vaccines in the short term. The exogenous factors in chicken embryo may also affect the quality of vaccine. Vero cells are recommended by WHO for vaccine production and can be cultured on a large scale using bioreactor. The influenza vaccine obtained on Vero cells can produce strong humoral and cellular immune responses. If a strain of H5N1 avian influenza vaccine cultured with Vero cells is developed, It is of great significance to control avian influenza. In this experiment, H5N1 avian influenza vaccine strain and H3N2 Vero adaptive strain selected by our laboratory were inoculated with chicken embryo for 24 hours to get allantoic fluid. The high yield adaptive strain of H5N1 Vero cells was obtained by adding the antiserum neutralization of H3N2 Vero cell acclimation strain to Vero cells. The strain was purified by plaque test. After 20 consecutive passages, the strain was identified by hemagglutination inhibition test and gene sequencing. The pathogenicity, immunogenicity and other biological characteristics of the H5N1 Vero cell adaptive strain were further determined. The obtained H5N1 Vero cell adaptive strain could be used in the development of human vaccine for H5N1 avian influenza.
【學位授予單位】：中國協(xié)和醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R392

【參考文獻】

相關期刊論文 前3條

1 ;Structure and Function of the NS1 Protein of Influenza A Virus[J];Acta Biochimica et Biophysica Sinica;2007年03期

2 靳毅，，武力，李益民，史晉芳，白植生;區(qū)帶超速離心提純流感病毒的初步探討[J];微生物學免疫學進展;1996年03期

3 龍進學;薛峰;彭宜;顧敏;劉秀梵;;H5N1亞型禽流感病毒NS第263~277位核苷酸缺失提高病毒對雞的致病力[J];微生物學報;2006年02期

本文編號：1691954