口蹄疫A型病毒AF/72株标准细胞毒株的研制  

Study of Reference Cellular Strain of AF/72 of FMD Type A Virus

作　　者：杜进鑫[1,2] 王永录[1] 张永光[1] 方玉珍[1] 蒋守田[1] 吕建亮[1] 潘丽[1] 刘力宽[1] 

机构地区：[1]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州730046 [2]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《华北农学报》2009年第3期74-77,共4页Acta Agriculturae Boreali-Sinica

基　　金：国家支撑计划项目(2006BAD06A06)

摘　　要：用口蹄疫A型病毒AF/72株的第3代乳鼠组织毒,通过乳鼠继续适应3代后,转适BHK-21细胞,获得该毒株的细胞毒AF/72/MF6/BF12,经测定其TCID50为10-8.0/mL;经RT-PCR获得其VP1基因序列,并与GenBank中的其他6株口蹄疫A型病毒株比对,同源性均大于85%;经无菌检验和外源病毒检验,纯净性达到兽用生物制品标准要求;经间接夹心ELISA测定,OD值均大于0.2,且该毒仅能被口蹄疫A型标准血清中和,具有型特异性;经紫外分光光度法测定其146S含量,均值为189 ng/mL,远大于22 ng/mL的国际标准。综合纯净性检验结果、特异性检验结果和146S含量,确定AF/72/MF6/BF12为口蹄疫A型病毒株AF/72的标准细胞毒。Get the foot-and-mouth virus AF/72/MF6/BF12 by rat rejuvenation and BHK-21 cells adaption using FMD virus AF/72/MF3 the use of Karber method to measure AF/72/MF6/BF12TCID50. its value was 8.0;the main antigen VP1 gene sequence was got by RT-PCR with 6 type A strains of FMDV in GenBank, homology was higher than 85 % . By the sterile examination, other exogenous virus test, its purity of AF/72/MF6/BF12 conform to Biological Products Standards for Animal Use;Indirect Sandwich ELISA OD Values greater than 0.2, This virus was only neutralized by the reference serum of FMD type A in line with the specific; 146S antigen content was measured by ultraviolet spectrophotometer, The average value was much higher than 22 ng/mL of international standards. AF/72/MF6/BF12 was identified as reference cellular strain of AF/72 of FMD type A virus integrating many parameters.

关 键 词：口蹄疫A型病毒 FMD 毒株 

分 类 号：Q78[生物学—分子生物学]