猪繁殖与呼吸综合征病毒NADC30-like株的分离鉴定及基因组分析OA北大核心CSTPCD
Isolation,Identification and Genomic Analysis of Porcine Reproductive and Respiratory Syndrome Virus NADC30-like Strain
猪繁殖与呼吸综合征(PRRS)呈现全球流行,对养猪行业造成巨大的经济损失.为增加PRRSV分离细胞种类的多样性,同时了解新疆地区猪繁殖与呼吸综合症病毒(PRRSV)遗传变异的现状,采用新鲜制备猪骨髓细胞诱导分化成猪骨髓源巨噬细胞,对新疆地区某猪场疑似患有PRRS的肺脏,接种至猪骨髓源巨噬细胞,分离获得1株PRRSV毒株,命名为XJYQ-2021.通过测序得到全基因组序列,对该毒株NSP2氨基酸序列缺失、ORF5及全基因组序列进行同源性、遗传进化及重组分析;利用间接免疫荧光试验进行鉴定,同时对第10、15、20代病毒液绘制生长曲线.结果显示,诱导分化的猪骨髓源巨噬细胞纯度高达90%以上,XJYQ-2021分离株在该细胞生长良好,可见明显细胞病变;通过生长曲线可观察到病毒含量随代次增加而递增,感染48 h左右病毒滴度达到峰值;该分离株基因组全长15 030 nt,与美洲型NADC30同源性为90%;XJYQ-2021分离株与VR2332相比,NSP2氨基酸序列与美洲型NADC3 0具有相同不连续缺失(111+1+19);全基因重组分析结果表明,XJYQ-2021分离株是重组病毒,有一个重组片段,其中NADC30是亲本毒株,与JXA1之间发生重组获得.研究结果可为PRRSV的病毒分离提供优选细胞种类,同时掌握了新疆地区PRRSV流行毒株的遗传进化现状.
Porcine reproductive and respiratory syndrome(PRRS)is becoming a global epidemic,causing significant economic losses to the global pig industry.In order to improve the selection of PRRSV isolation cell types and understand the current genetic variation status of porcine reproductive and respiratory syn-drome virus(PRRSV)in Xinjiang,this study used freshly prepared pig bone marrow cells to induce differ-entiation into porcine bone marrow derived macrophages.A PRRSV virus strain named XJYQ-2021 was i-solated from the lungs of a suspected PRRSV infected pig farm in Xinjiang,and the entire genome was ob-tained through sequencing,homology,genetic evolution,and recombination analysis on the NSP2 amino acid sequence deletion,ORF5,and whole genome sequence of the virus strain;Identification was conducted using indirect immunofluorescence assay,and growth curves were plotted for the 10th,15th,and 20th gen-eration virus solutions.The results showed that the purity of induced differentiation of porcine bone mar-row-derived macrophages reached over 90%,and the XJYQ-2021 isolate grew well in this cell type,show-ing obvious cytopathic effects;Through the growth curve,it can be observed that the virus content increa-ses with the increase of generations,and the virus titer reaches its peak around 48 hours after infection;The total genome length of this isolate is 15 030 nt,with 90%homology with the American type NADC30;Compared with VR2332,the XJYQ-2021 isolate showed the same discontinuous deletion(111+1+19)in the amino acid sequence of NSP2 as the American type NADC30;The results of whole gene recombination analysis indicated that the XJYQ-2021 isolate is a recombinant virus with a recombinant fragment,among which NADC30 is a parent virus strain that was obtained through recombination with JXA1.The results of this study provide optimal cells for the isolation of PRRSV virus,as well as the genetic evolution of PRRSV epidemic strains in the Xinjiang region.
毛丽萍;潘晓梅;曹剑;李俊辉;周涛;侯红娟;郑学功;贺笋
天康制药股份有限公司,新疆乌鲁木齐 830011
畜牧业
猪繁殖与呼吸综合征病毒类NADC30猪骨髓源巨噬细胞Nsp2重组分析
PRRSVNADC30-likepig bone marrow-derived macrophageNsp2recombination analysis
《动物医学进展》 2024 (007)
11-17 / 7
丝绸之路经济带创新驱动发展试验区乌昌石国家自主创新示范区科技发展计划项目(2022LQ03022)
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