摘要
为了解2014年云南省香猪群体中猪繁殖与呼吸综合征病毒(PRRSV)的流行特征,将疑似猪繁殖与呼吸综合征(PRRS)病例肺脏和淋巴结研磨处理后接种Marc-145细胞进行盲传,传至第2代48 h时出现2株病毒致Marc-145细胞聚集融合、巨细胞等病毒致细胞病变(CPE)效应现象,将其命名为YN-2XZ1和YN-3XZ2。细胞冻融3次后,使用Reed-Muench法计算病毒的半数感染力(TCID_(50))分别为10^(6.75),10^(6.05)TCID50/100μL。对ORF7基因进行序列测定、分析及N蛋白的免疫印迹(WB)检测,并与美洲经典毒株VR-2332、国内2001-2014年部分序列进行比对结果显示,本次获得的2株PRRSV之间核苷酸同源性为100%,与国内流行的变异毒株进化同步,说明目前云南香猪群中PRRSV以变异毒株为主,可能不具有高暴发性和高致病力,不会引起重大经济损失和社会恐慌,但仍需对此加以关注。通过磷酸化分析得出,暴发HP-PRRSV前N蛋白35位丝氨酸磷酸化概率67.4%~99.8%,暴发HP-PRRSV后N蛋白35位丝氨酸磷酸化概率低于50%,提示该位点氨基酸不易于磷酸化可能与NSP-2蛋白基因缺失从而引起HP-RRSV有关,需进一步研究验证。
To understand the epidemic characteristics of respiratory syndrome virus(PRRSV)in pig in Yunnan Province in 2014,the lung and lymph nodes of pigs of suspected cases of porcine reproductive and respiratory syndrome(PRRS)were ground and inoculated Marc-145 cells for blind transmission.At 48 h post inoculation,two strains of virus were found to cause cytopathic effects(CPE)such as aggregation and fusion of Marc-145 cells,they were named YN-2 XZ1 and YN-3 XZ2,respectively.After cells were treated with three freeze-thawing cycles,the 50%tissue culture infective dose(TCID50)was calculated by Reed-Muench method,it was 10^(6.75)TCID50/100μL and 10^(6.05)TCID_(50)/100μL,respectively.The N protein of the virus was detected by Western blot(WB),and the viral ORF7 gene was sequenced and analyzed,and the sequence was compared with the reference American strain VR-2332 and the partial sequence of domestic strains from 2001 to 2014.The results showed that the nucleotide homology between the two PRRSV strains was 100%,which was in step with the evolution of the mutant strains prevalent in China,indicating that PRRSV in Yunnan Xiang pig population are mainly variant strains,it may not have high pathogenicity and outbreak risk,and will not cause great economic loss and social panic,but it still needs to be paid attention to.The results of phosphorylation analysis showed that the probability of serine phosphorylation at position 35 of N protein was 67.4%-99.8%before outbreak of HP-PRRSV,and less than 50%after outbreak of HP-PRRSV,suggesting that this amino acid is not easy to be phosphorylated,which may be related to the deletion of gene of nsp-2 protein resulting in HPPRRSV,but this needs further research and verification.
作者
郑龙龙
朱玲云
刘萍丹
赵德育
刘佳
毕峻龙
杨贵树
舒相华
ZHENG Longlong;ZHU Lingyun;LIU Pingdan;ZHAO Deyu;LIU Jia;BI Junlong;YANG Guishu;SHU Xianghua(College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Province Center for Disease Control and Prevention in Chuxiong,Chuxiong,Yunnan 675000,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2021年第1期13-19,共7页
Chinese Journal of Veterinary Science
基金
云南省公益性关键技术研究开发计划子项目(2013CH001)。
