一株1型非洲马瘟疫苗毒种的初步鉴定

Preparation and Primary identification of One African Horse Sickness Virus Serotype 1 Vaccine Seed

关键词:  非洲马瘟病毒  血清1型  VERO细胞  疫苗种毒

Key words:  AHSV, Serotype 1, Vero, Vaccine seed

摘要:

为做好非洲马瘟疫苗的储备性研究,以更好应对我国周边地区非洲马瘟疫情传入风险,本研究对中国兽医微生物菌种保藏中心的一株1型非洲马瘟病毒鼠脑组织毒进行了复壮和细胞适应型培养,并对其病毒含量、特异性、对小鼠毒力等进行测定。结果显示,该毒种在VERO细胞连续传代至F10代后病变产生明显,病毒含量可稳定在10^6.5TCID50/0.1mL以上；对小鼠毒力表现稳定,细胞适应毒也可达10^6.5LD50/0.1mL。根据OIE参考引物进行RT-PCR鉴定,序列比对证实该毒株为血清1型,与泰国毒株THA2020/01拥有99.7%的同源性。血清学试验结果表明可被1型特异性血清中和。本研究获得了一株VERO细胞适应毒,可作为良好的疫苗毒株候选株,适用于规模化生产,为疫苗储备奠定良好基础。

Abstract:

In order to better tackle with the risk of introduction of African Horse Sickness from neighboring countries and to maintain the vaccine emergency stockpile, one African Horse Sickness Virus (AHSV) serotype 1 vaccine seed was prepared by rejuvenationSand adaptation to VERO cells, and its characteristics, viral titers, virulence in mice and specificity were determined. Results showed that this seed strain has a good characteristic. Virus titers could reach 10^6.5 TCID50/0.1mL, as well as the virulence in mice reaching 10^6.5LD50/0.1mL. Furthermore, sequence and alignment analysis indicated that the strain shares 99.7% homologySof nucleotide sequence with isolate THA2020/01, and serum neutralization test showed that it could be neutralized by specific serum, suggesting this prepared culture could be a good candidate as a vaccine strain, laying the foundation for subsequent research.