口蹄疫A型病毒AF/72株标准细胞毒株的研制

doi:10.7668/hbnxb.2009.03.016

华北农学报 ›› 2009, Vol. 24 ›› Issue (3): 74-77. doi: 10.7668/hbnxb.2009.03.016

口蹄疫A型病毒AF/72株标准细胞毒株的研制

杜进鑫¹, 王永录², 张永光², 方玉珍², 蒋守田², 吕建亮², 潘丽², 刘力宽²

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

收稿日期:2009-03-01 出版日期:2009-06-28
通讯作者: 王永录(1964 - ),男,甘肃临洮人,研究员,博士,主要从事动物分子免疫学和分子生物学研究。
作者简介:杜进鑫(1982-), 男, 甘肃靖远人, 在读硕士, 主要从事动物病毒分子免疫学研究.
基金资助:
国家支撑计划项目(2006BAD06A06)

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

DU Jin xin¹, ², WANG Yong lu², ZHANG Yong guang², FANG Yu zhen², JIANG Shou tian², LU Jian liang², PAN Li²

1. K ey Laboratory of Animal Virology of Agriculture,State K ey Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute,Chinese Academy of Agriculture Sciences,Lanzhou 730046, China;
2. College of Veterinary Medicine,G ansu Agricutural University,G ansu 730070,China

Received:2009-03-01 Published:2009-06-28

摘要/Abstract

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VPI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16S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16S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.

关键词: 口蹄疫A型病毒, FMD, 毒株

Abstract: Get the foot2and2mouth virus AF/ 72/ MF₆/BF₁₂by rat rejuvenation and BHK 221 cells adaption using FMD virus AF/ 72/ MF3the use of Karber method to measure AF/ 72/ MF₆/BF₁₂TCID50. its value was 8. 0;the main antigen VP₁ gene sequence was got by RT2PCR with 6 type A strains of FMDV in GenBank,homology was higher than 85 %.By the sterile examination,other exogenous virus test,its purityof AF/ 72/ MF₆/BF₁₂conform 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/BF₁₂was identified as reference cellular strain of AF/ 72 of FMD type A virus integrating many parameters.

Key words: FMD type A virus, FMD, Strain

中图分类号:

杜进鑫, 王永录, 张永光, 方玉珍, 蒋守田, 吕建亮, 潘丽, 刘力宽. 口蹄疫A型病毒AF/72株标准细胞毒株的研制[J]. 华北农学报, 2009, 24(3): 74-77. doi: 10.7668/hbnxb.2009.03.016.

DU Jin xin,, WANG Yong lu, ZHANG Yong guang, FANG Yu zhen, JIANG Shou tian, LU Jian liang, PAN Li, LIU Li kuan. Study of Reference Cellular Strain of AF/72 of FMD Type A Virus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(3): 74-77. doi: 10.7668/hbnxb.2009.03.016.

http://www.hbnxb.net/CN/Y2009/V24/I3/74

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

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