【摘要】：為了解類NADC30豬繁殖與呼吸綜合征病毒(PRRSV)在我國新的流行特點,本研究根據(jù)GenBank中美國2012年提交的PRRSV NADC30株全基因組序列設(shè)計合成8對引物,通過RT-PCR的方法對本實驗室2016年從4個省搜集的臨床樣品中檢測到的15株類NADC30 PRRSVs株進(jìn)行了全基因組測序,并且結(jié)合參考株進(jìn)行了序列分析。NSP2推導(dǎo)氨基酸的序列比對表明,15株P(guān)RRSVs均具有aa323~aa433+aa484+aa505~aa523(111+1+19)131個氨基酸缺失的分子特征;經(jīng)RDP4和Simplot生物學(xué)軟件分析表明,15株P(guān)RRSVs中14株為重組病毒株,參與重組的親本病毒株為PRRSV NADC30株,提供重組基因片段的病毒株為HP-PRRSV或以VR2332為代表的經(jīng)典PRRSV株,這些類NADC30重組病毒株的重組位置、重組的基因片段及重組片段長度均具有多樣性,而且重組的位置多位于編碼非結(jié)構(gòu)蛋白或次要結(jié)構(gòu)蛋白的基因區(qū)域;全基因組遺傳演化分析表明,這些病毒株遠(yuǎn)離國內(nèi)之前流行的HP-PRRSV,同源性僅為83.4%~87.2%,而與其它的類NADC30病毒株位于一個相對獨立的分支,以上結(jié)果均表明這類病毒株已形成了一個新的亞群,命名為5亞群。本研究中不同地區(qū)PRRSV的核苷酸同源性差異較大,僅為88.6%~99.6%,并且只有2個病毒株可以感染PAM和MARC-145細(xì)胞。本研究表明,以基因缺失和基因重組多樣性為主要特征的5亞群PRRSV已在我國流行,其生物學(xué)特性有待進(jìn)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.
【作者單位】： 中國農(nóng)業(yè)科學(xué)院哈爾濱獸醫(yī)研究所;山東信得生物科技有限公司;南陽師范學(xué)院;中國農(nóng)業(yè)科學(xué)院上海獸醫(yī)研究所;
【基金】：國家自然科學(xué)基金(31270045);國家自然科學(xué)基金(31502097)
【分類號】：S852.65

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