猪伪狂犬病病毒变异株传代致弱JS18-150株的获得与鉴定
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作者单位:

武汉科前生物股份有限公司,武汉 430206

作者简介:

侯真真,E-mail:2265446230@qq.com

通讯作者:

徐高原,E-mail:xugaoyuan@163.com

中图分类号:

S852.651

基金项目:

湖北省重点研发计划项目(2023BBB177)


Obtaining and identifying a mutated and passage-attenuated strain JS18-150 of porcine pseudorabies virus
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Affiliation:

Wuhan Keqian Biology Co., Ltd., Wuhan 430206, China

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    摘要:

    为获得安全性良好、免疫原性高的变异株弱毒苗,采用鸡胚成纤维细胞(CEF)传代培养的方法对猪伪狂犬病病毒流行毒株JS18株进行传代,最终获得JS18-150株,随后通过PCR鉴定、测序、致病力试验和免疫效力试验对JS18-150株的体外生长特性、安全性和免疫原性进行初步研究。试验结果显示,在CEF上连续传代后,JS18株的gIgEUS9基因缺失,获得的JS18-150株毒力已致弱,在CEF上适应性显著提高,与JS18株生长动力学相似;将JS18-150株以107.0 TCID50/mL接种仔猪,所有仔猪均未观察到任何临床症状,且体温均未超过40.5 ℃;JS18-150株以106 TCID50/mL接种仔猪后能保护仔猪有效抵御JS18株和HBZL05株的攻击。结果表明,JS18-150株对仔猪安全性良好,可作为PRV弱毒疫苗研制的候选毒株。

    Abstract:

    Pseudorabies is an acute infectious disease of pigs, which is caused by the infection of pseudorabies virus(PRV). Pseudorabies is mainly prevented by vaccine, and PRV gene-deleted vaccine is the most widely used and effective. Since 2011, a new outbreak has occurred in many pig farms vaccinated with the Bartha-K61 vaccine and has spread to farms in most parts of the country, causing enormous economic losses to the development of our farms. The study found that the new outbreak was caused by PRV variants and showed that the commercially vaccine did not provide complete protection. Therefore, it is urgent to develop a new and efficient PRV vaccine.The prevalent strain JS18 of porcine pseudorabies virus was passaged with chicken embryo fibroblast (CEF) cells to obtain the attenuated vaccine with good safety and high immunogenicity for mutated strain of porcine pseudorabies virus. JS18-150 strain was obtained. Preliminary studies were conducted on the in vitro growth characteristics, safety, and immunogenicity of JS18-150 strain through PCR identification, sequencing, pathogenicity testing, and immunogenicity testing. The results showed that the gIgE, and US9 genes of JS18 strain were deleted and the virulence of JS18-150 strain obtained was weakened after continuous passage on CEF. The adaptability on CEF of JS18-150 strain obtained was significantly improved. The growth kinetics of JS18-150 strain obtained was similar to that of the original JS18 strain. No clinical symptoms were observed in all piglets inoculated with 107.0 TCID50 JS18-150 strain obtained, and the rectal temperature did not exceed 40.5 ℃. Inoculation with 106.0 TCID50 JS18-150 strain obtained effectively protected piglets from the attacks of JS18 strain and HBZL05 strain. It is indicated that JS18-150 strain obtained has good safety for piglets and can be used as a candidate strain for the development of attenuated PRV vaccines.

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侯真真,张华伟,郝根喜,罗修鑫,宋文博,周明光,陈焕春,徐高原.猪伪狂犬病病毒变异株传代致弱JS18-150株的获得与鉴定[J].华中农业大学学报,2024,43(5):178-185

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  • 收稿日期:2023-10-24
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  • 在线发布日期: 2024-10-08
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