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当前位置: 首页 > 文章 > 5株猫杯状病毒中国分离株的细胞适应性及遗传进化分析 经济动物学报 2016,20 (1) 8-12
Position: Home > Articles > Cell Passage of Five Feline calicivirus Strains Isolated in China and Phylogenetic Analysis Journal of Economic Animal 2016,20 (1) 8-12

5株猫杯状病毒中国分离株的细胞适应性及遗传进化分析

作  者：
应瑛;王一鸣;马剑钢;曹东梅;李响;高玉伟;杨松涛;胡桂学
单  位：
吉林农业大学动物科学技术学院;军事医学科学院军事兽医研究所
关键词：
猫杯状病毒;F81细胞;适应性传代
摘  要：
将CH-JL1、CH-JL2、CH-JL3、CH-JL4、CH-SH 5株猫杯状病毒中国分离株分别在F81细胞上连续传至15代。细胞传代前,测定病毒的细胞半数感染量(TCID_(50)),再以0.1 MOI的感染复数接种细胞。采用RTPCR方法扩增FCV各代次的ORF2基因片段并测序,用DNASTAR和MEGA软件分析其氨基酸变异和遗传进化特点。试验结果显示,细胞适应传代后,FCV各代次毒株滴度逐渐稳定,均位于10~(-9)TCID_(50)/mL~10~(-10)。TCID_(50)/mL之间。ORF2基因测序表明,FCV传代后氨基酸变异位点包括CH—JL1的P57S和G494R、CH-JL3的S440L和N519D、CH—JL4的V614I、CH—SH的K448R。遗传进化分析表明CH—JL2和CH-JL3处于同一进化分支上,同源性为95.9%;CH-JL1和国内FB-NJ-13株、CH-SH和国内GD株分别在一个进化分支,CH-JL4和国外F65株遗传关系较近。5株猫杯状病毒在F81细胞传代后滴度较高,稳定性良好,为将来FCV疫苗株的研发奠定了基础。
译  名：
Cell Passage of Five Feline calicivirus Strains Isolated in China and Phylogenetic Analysis
作  者：
YING Ying;WANG Yiming;MA Jiangang;CAO Dongmei;LI Xiang;GAO Yuwei;YANG Songtao;HU Guixue;College of Animal Science and Technology,Jilin Agricultural University;Institute of Military Veterinary Medicine,Academy of Military Medical Sciences;
关键词：
Feline calicivirus;;F81 cells;;adaptive passage
摘  要：
The five FCV strains,namly CH-JL1,CH-JL2,CH-JL3,CH-JL4 and CH-SH,isolated from cats were cultivated continuingly in F81 cells for 15 passages.After measurement of TCID_(50),the F81 cells were infected by different strains at a MOI of 0.1.In addition,the ORF2 genes of these passages were amplified by RT—PCR method and the sequence alignment were analyzed.We used DNASTAR or MEGA software to align amino acid variation and phylogenetic trees.The five FCV strains were adapted in F81 cells for 15 passages and the titers were increased steadily between 10~(-9) TCID_(50)/mL and 10~(-10) TCID_(50)/mL.The sequencing result of 0RF2 showed some differences in nucleotides and amino acids.The variations of amino acid were located in P57 S,G494R of CH-JL1,S440 L and N519 D of CH-JL3,V614 I of CH-JL4 and K448 R of CH-SH.Phylogenetic analysis based on the nucleotide of 0RF2 genes showed that CH-JL2 and CH-JL3 belonged to the same branch on phylogenetic tree and the homology of nucleotide sequences was over 99%.Others were separated in a branch,such as CH-JL1 and FB-NJ-13,CH-JL4 and F65,CH-SH and GD.The five FCV strains could steadily replicated to high titers in F81 cells,on which laid a foundation for development of FCV vaccines.

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