Obtaining and identifying a mutated and passage-attenuated strain JS18-150 of porcine pseudorabies virus
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Wuhan Keqian Biology Co., Ltd., Wuhan 430206, China

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S852.651

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    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.

    Table 1 Primers used for PCR
    Table 4 Protective efficacy of JS18-150 immunization
    Table 2 PRV gB and gE-specific antibody in piglets inoculated with different passaged PRV
    Fig.1 CPE caused by PRV JS18 in CEF
    Fig.2 Plaque morphology of PRV JS18 and JS18-150 in CEF
    Fig.3 Growth curves of PRV JS18 and JS18-150
    Fig.4 Survival rates of piglets infected with different passages of PRV
    Fig.5 Protective efficacy of JS18-150 immunization
    Fig.6 Autopsy and microscopic examination of brain tissue after challenge(400×)
    Fig.7 Autopsy and microscopic examination of tonsil tissue after challenge(400×)
    Table 3 Safety of piglets vaccinated with JS18-150
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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. Obtaining and identifying a mutated and passage-attenuated strain JS18-150 of porcine pseudorabies virus[J]. Jorunal of Huazhong Agricultural University,2024,43(5):178-185.

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  • Received:October 24,2023
  • Online: October 08,2024
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