史爱华;姜北宇;沈佳;景小冬;章振华;李林;张建伟;

• 1:北京市农林科学院畜牧兽医研究所

摘要(Abstract)：

[目的]探讨不同胰蛋白酶浓度对低致病力禽流感病毒在MDCK细胞上增殖后的HA滴度。[方法]通过3株禽流感H9亚型病毒分别接种MDCK细胞单层,加入含有不同胰蛋白酶浓度的DMEM维持液,每隔24 h观察细胞病变,并测定上清液中的HA滴度。[结果]当维持液中胰蛋白酶含量为10～20μg/ml时,培养液上清液中的HA滴度最高,达到7 log2(1∶128)。当病毒的接种浓度为10-3和10-4时,MDCK细胞在96 h几乎全部病变,峰值出现在接种后的72～96 h。[结论]当维持液中胰蛋白酶含量为10～20μg/ml时,有利于H9亚型AIV病毒在MDCK细胞上增殖。

关键词(KeyWords)： H9N2亚型禽流感病毒;MDCK细胞;胰蛋白酶浓度

Abstract:

Keywords:

基金项目(Foundation): 北京市农林科学院资助项目(2010A007)

作者(Author): 史爱华;姜北宇;沈佳;景小冬;章振华;李林;张建伟;

Email:

DOI: 10.13989/j.cnki.0517-6611.2011.35.223

参考文献(References)：

• [1]毕英佐.H9N2亚型禽流感的流行现状和防制措施[J].中国家禽,2009,31(11):33-34.
• [2]周蛟.世界主要禽病流行现状[J].中国家禽,2007,29(4):1-4.
• [3]叶柱德,胡江锋.H9N2亚型禽流感病毒疫苗研究进展[J].动物医学进展,2008,29(2):89-92.
• [4]KISTNER O,BARRETT PN,MUNDT W,el al.Development of a mammali-an cell(Vero)derived candidate influenza virus vaccine[J].Vaccine,1998,16(9/10):960-968.
• [5]KEIICHI MAKIZUMI,KAZUHIKO KIMACHI,KATSUHIKO FUKADA,etal.Timely production of A/Fujian-like influenza vaccine matching the2003-2004 epidemic strain may have been possible using Madin-Darbycanine kidney cells[J].Vaccine,2008,26:6852-6858.
• [6]LOHR V,GENZEL Y,BEHRENDT I,et al.A new MDCK suspension linecultivated in a fully defined medium in stirred-tank and wave bioreactor[J].Vaccine,2010,28:62560-6264.
• [7]YOUIL R,SU Q,TONER T J,et al.Comparative study of influenza virusreplication in Vero and MDCK cell lines[J].J Virol Methods,2004,120(1):23-31.
• [8]HALPERIN S A,SMITH B,MABROUK T,et al.Safety and immunogenici-ty of a trivalent,inactivated,mammalian cell culture derived influenza vac-cine in healthy adults,seniors,and children[J].Vaccine,2002,20(7/8):1240-1247.
• [9]JONATHAN LIU,XIAO SHI,RICHARD SCHWARTZ,et al.Use of MDCKcells for production of live attenuated influenza vaccine[J].Vaccine,2009,27:6460-6463.
• [10]DONG J,ROTH M G,HUNTER E.A chimeric avian retrovirus containingthe influenza virus hemagglutinin gene has an expanded host range[J].JVirol,1992,66:7374-7382
• [11]刘丽玲,王秀荣,陈化兰,等.不同亚型禽流感病毒在MDCK细胞上的生长特性[J].中国兽医杂志,2006,42(10):18-20.