2016年我国部分地区类NADC30新亚群猪繁殖与呼吸综合征病毒的遗传变异分析
发布时间：2018-07-27 14:55
【摘要】：为了解类NADC30猪繁殖与呼吸综合征病毒(PRRSV)在我国新的流行特点,本研究根据GenBank中美国2012年提交的PRRSV NADC30株全基因组序列设计合成8对引物,通过RT-PCR的方法对本实验室2016年从4个省搜集的临床样品中检测到的15株类NADC30 PRRSVs株进行了全基因组测序,并且结合参考株进行了序列分析。NSP2推导氨基酸的序列比对表明,15株PRRSVs均具有aa323~aa433+aa484+aa505~aa523(111+1+19)131个氨基酸缺失的分子特征;经RDP4和Simplot生物学软件分析表明,15株PRRSVs中14株为重组病毒株,参与重组的亲本病毒株为PRRSV NADC30株,提供重组基因片段的病毒株为HP-PRRSV或以VR2332为代表的经典PRRSV株,这些类NADC30重组病毒株的重组位置、重组的基因片段及重组片段长度均具有多样性,而且重组的位置多位于编码非结构蛋白或次要结构蛋白的基因区域;全基因组遗传演化分析表明,这些病毒株远离国内之前流行的HP-PRRSV,同源性仅为83.4%~87.2%,而与其它的类NADC30病毒株位于一个相对独立的分支,以上结果均表明这类病毒株已形成了一个新的亚群,命名为5亚群。本研究中不同地区PRRSV的核苷酸同源性差异较大,仅为88.6%~99.6%,并且只有2个病毒株可以感染PAM和MARC-145细胞。本研究表明,以基因缺失和基因重组多样性为主要特征的5亚群PRRSV已在我国流行,其生物学特性有待进一步研究。
[Abstract]:In order to understand the new epidemic characteristics of NADC30 like porcine reproductive and respiratory syndrome virus (PRRSV) in China, eight pairs of primers were designed and synthesized according to the whole genome sequence of PRRSV NADC30 strain submitted by the United States in 2012 in GenBank. A total of 15 NADC30 PRRSVs-like strains from clinical samples collected in 4 provinces in 2016 were sequenced by RT-PCR. The sequence alignment of amino acids deduced from NSP2 combined with reference strain showed that all of the 15 PRRSVs strains had the molecular characteristics of aa323~aa433 aa484 aa505~aa523 (111119) 131 amino acid deletion, and 14 of the 15 PRRSVs strains were recombinant viruses by RDP4 and Simplot analysis. The parent of the virus involved in the recombination was PRRSV NADC30 strain, the strain providing recombinant gene fragment was HP-PRRSV or the classical PRRSV strain represented by VR2332. The recombination location, the length of the recombinant gene fragment and the recombinant fragment length of these NADC30 like recombinant virus strains were all diverse. Moreover, the position of recombination is mostly located in the gene region that encodes non-structural protein or minor structural protein, and the genetic evolution analysis of the whole genome shows that, These strains are far away from the HP-PRRSVs which were prevalent in China, and the homology is only 83.4% and 87.2%, while they are located in a relatively independent branch with other NADC30 like virus strains. All the above results indicate that this kind of virus strain has formed a new subgroup, named 5 subgroup. In this study, the nucleotide homology of PRRSV in different regions was significantly different, which was only 88.60.99. 6 and only two strains of the virus could infect PAM and MARC-145 cells. This study shows that subgroup 5 PRRSV, which is characterized by gene deletion and gene recombination diversity, has been prevalent in China, and its biological characteristics need to be further studied.
【作者单位】： 中国农业科学院哈尔滨兽医研究所;山东信得生物科技有限公司;南阳师范学院;中国农业科学院上海兽医研究所;
【基金】：国家自然科学基金(31270045);国家自然科学基金(31502097)
【分类号】：S852.65
[Abstract]:In order to understand the new epidemic characteristics of NADC30 like porcine reproductive and respiratory syndrome virus (PRRSV) in China, eight pairs of primers were designed and synthesized according to the whole genome sequence of PRRSV NADC30 strain submitted by the United States in 2012 in GenBank. A total of 15 NADC30 PRRSVs-like strains from clinical samples collected in 4 provinces in 2016 were sequenced by RT-PCR. The sequence alignment of amino acids deduced from NSP2 combined with reference strain showed that all of the 15 PRRSVs strains had the molecular characteristics of aa323~aa433 aa484 aa505~aa523 (111119) 131 amino acid deletion, and 14 of the 15 PRRSVs strains were recombinant viruses by RDP4 and Simplot analysis. The parent of the virus involved in the recombination was PRRSV NADC30 strain, the strain providing recombinant gene fragment was HP-PRRSV or the classical PRRSV strain represented by VR2332. The recombination location, the length of the recombinant gene fragment and the recombinant fragment length of these NADC30 like recombinant virus strains were all diverse. Moreover, the position of recombination is mostly located in the gene region that encodes non-structural protein or minor structural protein, and the genetic evolution analysis of the whole genome shows that, These strains are far away from the HP-PRRSVs which were prevalent in China, and the homology is only 83.4% and 87.2%, while they are located in a relatively independent branch with other NADC30 like virus strains. All the above results indicate that this kind of virus strain has formed a new subgroup, named 5 subgroup. In this study, the nucleotide homology of PRRSV in different regions was significantly different, which was only 88.60.99. 6 and only two strains of the virus could infect PAM and MARC-145 cells. This study shows that subgroup 5 PRRSV, which is characterized by gene deletion and gene recombination diversity, has been prevalent in China, and its biological characteristics need to be further studied.
【作者单位】： 中国农业科学院哈尔滨兽医研究所;山东信得生物科技有限公司;南阳师范学院;中国农业科学院上海兽医研究所;
【基金】：国家自然科学基金(31270045);国家自然科学基金(31502097)
【分类号】：S852.65
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