猪丁型冠状病毒的分离鉴定

Isolation and Identification of Porcine Deltaconoravirus

2016年从某商品化猪场采集到6份疑为病毒性腹泻的腹泻仔猪小肠样本,利用RT-PCR对样品进行检测,检出1份猪丁型冠状病毒(Porcine Deltaconoravirus,PDCoV)阳性样品,使用ST细胞进行病毒分离和传代。通过细胞病变(CPE)、RT-PCR及S基因序列测定分析对分离的病毒进行鉴定,试验结果表明,已成功分离到一株能在ST细胞上稳定增殖并能产生典型CPE的猪丁型冠状病毒,命名为SCMS株。该毒株S基因长度为3480个核苷酸,与GenBank中登录23个参考毒株之间的同源性为95.9%~98.5%,其中与2018年山东分离株SD-11-2018同源性最高,为98.5%。该毒株的成功分离为进一步开展PDCoV的生物学特性研究和疫苗研制奠定了基础。

In 2016,the six small intestine samples of diarrheic piglets suspected of viral diarrhea were collected from a commercial pig farm.One sample was verified as PDCoV positive by RT-PCR.The virus was isolated on ST cell line.Then the isolated virus was identified by CPE,RT-PCR and sequence analysis of spike gene.The results showed that a strain of PDCoV named SCMS was successfully isolated.The spike gene sequence of PDCoV SCMS was 3480 nucleotides in length and shared 95.9%to 98.5%nucleotide identity with other 23 reference PDCoV strains.The highest sequence identity(98.5%)was found with SD-11-2018.This study would lay a foundation for further study on the biological characteristics and vaccine development of PDCoV.