猪流行性腹泻病毒YL112株的M基因序列分析及其在Vero细胞的传代适应性研究

天津农学院学报 ›› 2017, Vol. 24 ›› Issue (2): 34-38.

猪流行性腹泻病毒YL112株的M基因序列分析及其在Vero细胞的传代适应性研究

蓝日国¹, 张书晴¹, 张金秋², 苗晋锋^{1,通信作者}

1. 南京农业大学动物医学院,南京 210095;
2. 江苏省农业科学院国家兽医疫苗工程技术研究中心,南京 210014

收稿日期:2017-03-29 出版日期:2017-06-30 发布日期:2019-10-15
通讯作者: 苗晋锋（1976-）,男,河南新乡人,副教授,博士,主要从事动物健康调控的研究。E-mail：mjf171647@126.com。
作者简介:蓝日国（1993-）,男,广西梧州人,本科在读,主要从事动物健康调控的研究。
基金资助:
国家自然科学基金项目“线粒体MAVS 蛋白在抗H₃N₂亚型猪流感病毒感染过程中的作用”（31201873）; 国家自然科学基金项目“肌醇磷脂-Ca²⁺-NFAT途径在牛磺酸缓解乳房链球菌感染中的作用”（31672515）

Sequence Analysis of the M gene of Porcine Epidemic Diarrhea VirusYL112 Strain and Its Adaptation to Vero Cells

LAN Ri-guo¹, ZHANG Shu-qing¹, ZHANG Jin-qiu², MIAO Jin-feng^{1,Corresponding Author}

1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China;
2.National Research Center for Veterinary Vaccine Engineering and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2017-03-29 Online:2017-06-30 Published:2019-10-15

摘要/Abstract

摘要： 为了解PEDV YL112株与其他不同来源PEDV毒株之间在分子生物学上的差异,对PEDV YL112株的M基因进行了扩增和序列分析。结果表明：PEDV YL112株与参考株的M基因核苷酸同源性在97.9%~99.8%之间,显示出高度保守性。系统进化树分析表明,PEDV YL112株与经典株CV777的亲缘关系较远,而与我国地方流行毒株HLJBY分离株以及HN-XYYYP-2007的亲缘关系较近。为进一步了解该毒株的复制特征,采用Vero 细胞对PEDV YL112株进行体外细胞传代适应性研究。结果表明：经过连续传代培养后,PEDV YL112株对Vero细胞的适应性显著增强,其滴度从第5代的 10^4.8 TCID₅₀ /0.1mL 提高至第 40 代的 10^7.6 TCID₅₀ /0.1mL。本研究对于分析PEDV的遗传变异规律、了解病毒的致病特征及开发新型疫苗等均具有重要理论和现实意义。

关键词: PEDV YL112株, M基因, 序列分析, Vero细胞, 适应性

Abstract: To analyze the genetic variation between PEDV YL112 strain and other isolates, the M gene of PEDV YL112 strain was amplified and sequenced. The M gene of PEDV YL112 strain shared 97.9%-99.8% homology with PEDV reference strains. Phylogenetic analysis indicated that PEDV YL112 strain was genetically close to isolates HLJBY and HN-XYYYP-2007 rather than the classic strain CV777. Furthermore, adaptation of PEDV YL112 to Vero cells was conducted and its titer significantly increased from 10^4.8TCID₅₀ /0.1mL to 10^7.6TCID₅₀ /0.1mL by cellular adaptation. These results may contribute to further study on the genetic evolution, viral virulence and new vaccines of PEDV.

Key words: PEDV YL112, M gene, sequence analysis, Vero cells, adaptation

中图分类号:

S858.28

蓝日国, 张书晴, 张金秋, 苗晋锋. 猪流行性腹泻病毒YL112株的M基因序列分析及其在Vero细胞的传代适应性研究[J]. 天津农学院学报, 2017, 24(2): 34-38.

LAN Ri-guo, ZHANG Shu-qing, ZHANG Jin-qiu, MIAO Jin-feng. Sequence Analysis of the M gene of Porcine Epidemic Diarrhea VirusYL112 Strain and Its Adaptation to Vero Cells[J]. Journal of Tianjin Agricultural University, 2017, 24(2): 34-38.

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http://xuebao.tjau.edu.cn/CN/Y2017/V24/I2/34

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