【摘要】： 據(jù)我國CDC統(tǒng)計(jì),2008年我國人狂犬病發(fā)病數(shù)已突破3000例,正在形成第三次流行高峰。流行病學(xué)調(diào)查表明,95%以上的人狂犬病病例是因?yàn)榕c患病或帶毒的犬和貓直接或間接接觸引起的。因此,有效遏制人狂犬病,必須控制動(dòng)物狂犬病,研制安全有效的動(dòng)物狂犬疫苗,對(duì)狂犬病流行地區(qū)的犬、貓等動(dòng)物施行強(qiáng)制免疫接種。而疫苗的質(zhì)量,即疫苗的免疫原性和安全性是預(yù)防狂犬病的關(guān)鍵。為此,我們開展了狂犬病濃縮純化滅活疫苗的實(shí)驗(yàn)研究。 本研究首先對(duì)犬五聯(lián)弱毒疫苗中的狂犬病毒ERA株第15代Vero細(xì)胞培養(yǎng)毒Rb/E3-15進(jìn)行了鑒定。鑒定后的Rb/E3-15株,用Vero細(xì)胞大量增殖,經(jīng)醋酸鋅沉淀濃縮, Sepharose 4FF層析純化;通過BPL(β-丙內(nèi)酯)和甲醛的滅活比較實(shí)驗(yàn),選擇了低濃度BPL滅活抗原,配以合適比例的脂質(zhì)體和Al(OH)3佐劑,制備了狂犬病濃縮純化滅活佐劑疫苗。為了找到一種對(duì)狂犬疫苗抗體檢測(cè)的理想方法,我們分別用本實(shí)驗(yàn)室建立的熒光抗體病毒中和試驗(yàn)(FAVN)、快速熒光灶抑制試驗(yàn)和犬抗體檢測(cè)試劑盒(ELISA法)對(duì)實(shí)驗(yàn)室收集的狂犬疫苗免疫后的犬血清進(jìn)行測(cè)定比較,結(jié)果顯示FAVN與ELISA法、RFFIT法具有相似的敏感性與特異性。用制備的濃縮純化滅活佐劑疫苗免疫小鼠和犬,FAVN法測(cè)定免疫后抗體,結(jié)果顯示該濃縮純化滅活疫苗可誘導(dǎo)小鼠和犬產(chǎn)生高水平中和抗體,小鼠攻毒實(shí)驗(yàn)使小鼠獲得完全的免疫保護(hù);在犬體內(nèi)誘導(dǎo)的中和抗體水平明顯優(yōu)于濃縮純化前疫苗,犬的外周血淋巴細(xì)胞轉(zhuǎn)試驗(yàn)檢測(cè)表明,脂質(zhì)體佐劑疫苗的細(xì)胞免疫效果要優(yōu)于Al(OH)3佐劑疫苗。該濃縮純化滅活疫苗接種受試動(dòng)物后無不良反應(yīng),疫苗安全性好。
[Abstract]:According to CDC statistics, the number of rabies cases in China has exceeded 3000 cases in 2008, which is forming the third epidemic peak. Epidemiological survey showed that more than 95% of human rabies cases were caused by direct or indirect contact with sick or infected dogs and cats. Therefore, it is necessary to control animal rabies, develop a safe and effective animal rabies vaccine, and enforce vaccination against dogs and cats in rabies endemic areas. The quality of the vaccine, that is, the immunogenicity and safety of the vaccine, is the key to the prevention of rabies. Therefore, we carried out an experimental study on rabies concentrated and purified inactivated vaccine. In this study, the virus Rb/E3-15 of rabies virus ERA strain cultured in the 15th passage of Vero cells was first identified. The identified Rb/E3-15 strain was proliferated by Vero cells and concentrated by zinc acetate precipitation and purified by Sepharose 4FF chromatography. By comparing the inactivation of BPL (尾 -propionolactone) and formaldehyde, the inactivated antigen of low concentration BPL was selected, and the adjuvant of liposome and Al (OH) 3 was used to prepare the concentrated and purified adjuvant vaccine of rabies. In order to find an ideal method for detection of rabies vaccine antibody, we used fluorescent antibody neutralization test (FAVN),) established in our laboratory. The rapid fluorescent foci inhibition test and canine antibody test kit (ELISA) were used to detect and compare the serum of rabies vaccine immunized in laboratory. The results showed that FAVN had similar sensitivity and specificity to ELISA method and RFFIT method. Mice and dogs were immunized with concentrated and purified inactivated adjuvant vaccine. The antibody was determined by FAVN method. The results showed that the concentrated and purified inactivated vaccine could induce high level neutralizing antibody in mice and dogs. The mice were immunized completely by the virus attack test. The level of neutralizing antibody induced in dogs was significantly better than that of pre-concentrated and purified vaccines. The peripheral blood lymphocyte transposition test of dogs showed that the cellular immune effect of liposome adjuvant vaccine was better than that of Al (OH) _ 3 adjuvant vaccine. There was no adverse reaction after inoculation of the concentrated and purified inactivated vaccine, and the vaccine was safe.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李成文 ,陳詠梅;脂質(zhì)體佐劑疫苗研究進(jìn)展[J];國外醫(yī)學(xué).預(yù)防.診斷.治療用生物制品分冊(cè);2003年06期

2 張玉慧,高春潤(rùn),孫秀華,趙剛,潘廣域,王忠海,胡金東,秦賢;狂犬病疫苗純化工藝的建立[J];中國生物制品學(xué)雜志;1999年04期

3 董關(guān)木，劉增順，俞永新，嚴(yán)子林，劉景華;狂犬病毒CTN-1株在Vero細(xì)胞上的適應(yīng)傳代研究[J];微生物學(xué)免疫學(xué)進(jìn)展;1995年02期

4 高鴻瑞，魏至棟，單秀琴，，阮峰，趙新華，袁文潭，楊輝，李根明;狂犬病病毒Vero細(xì)胞適應(yīng)株的建立及其應(yīng)用于疫苗生產(chǎn)的可行性探討[J];微生物學(xué)免疫學(xué)進(jìn)展;1995年03期

5 李光譜,郭巖;預(yù)防性疫苗用于治療的研究[J];微生物學(xué)免疫學(xué)進(jìn)展;2004年01期

6 胡靜,王志高;江蘇省狂犬病40年流行特征分析[J];現(xiàn)代預(yù)防醫(yī)學(xué);1997年03期

7 唐青,趙秀芹,陶曉霞;中國人間狂犬病流行近況分析[J];中華流行病學(xué)雜志;2001年01期

8 林海祥;鋁佐劑對(duì)實(shí)驗(yàn)狂犬病疫苗的影響[J];中華實(shí)驗(yàn)和臨床病毒學(xué)雜志;1999年02期

9 李萍,朱家鴻,嚴(yán)家新,郭斐,胡巧玲,王繼麟,劉碧芬,阮力;狂犬病毒糖蛋白及核蛋白在非復(fù)制型痘苗病毒天壇株中的共同表達(dá)[J];中華微生物學(xué)和免疫學(xué)雜志;2000年05期

10 張守峰;曹亮;張菲;李海濤;李清竹;扈榮良;;狂犬病毒核蛋白單抗和熒光抗體中和試驗(yàn)的建立與應(yīng)用[J];中華檢驗(yàn)醫(yī)學(xué)雜志;2006年06期

本文編號(hào)：2317828