PRRSV广东株XH-GD的分离及其NSP2、ORF3、ORF5的序列分析

    Isolation and sequence analysis of NSP2, ORF3 and ORF5 of PRRSV XH-GD strain from Guangdong province

    • 摘要: 将采集的广东某猪场疑似猪高热症病料处理后接种于Marc-145细胞,出现细胞聚集、固缩、脱落等特异性细胞病变。根据GenBank中猪繁殖与呼吸综合征病毒(PRRSV)的基因序列设计并合成针对NSP2、ORF3和ORF5基因的引物,扩增目的基因并测序分析。结果表明:分离毒株XH-GD株PRRRSV属于美洲型,且NSP2基因发生30个氨基酸的不连续缺失;XH-GD的NSP2、ORF3和ORF5基因与HUB1、NX06尧BJsy06、VR-2332、HB-1(sh)2002、HB-2(sh)2002、CH-la和RespPRRS MLV等毒株等位基因的核苷酸同源性分别为83.3%~98.9%、88.6%~99.2%、88.1%~99.2%,推导的氨基酸同源性分别为76.8%~98.3%、83.7%~98.8%、88.1%~99.2%;而与LV的等位基因核苷酸同源性分别为52.9%、65.0%和64.3%,推导的氨基酸同源性分别为31.3%、57.5%和58.9%。基因系统发育树分析表明,XH-GD与NX06、BJsy06株的亲缘关系很近,与HUB1、NX06、BJsy06、VR-2332、HB-1(sh)2002、HB-2(sh)2002、CH-la和RespPRRSMLV亲缘关系较近,与LV毒株处于不同的分支。

       

      Abstract: The samples collected from Guangdong province were inoculated to Marc-145 cell, the PRRSV specific cytopathogenic effect (CPE) was observed and the isolates named XH-GD obtained. After reverse transcription polymerase chain reaction, the amplified NSP2、ORF3 and ORF5 fragments were analyzed by sequence comparison. The results demonstrated that XH-GD belonged to the North American genotype and its NSP2 had 30 amino acids deletion. The NSP2, ORF3 and ORF5 of XH-GD shared 83.3%~98.9%, 88.6%~99.2% and 88.1%~99.2% of nucleotide similarity, and 76.8%~98.3%, 83.7%~98.8% and 88.1%~99.2% of deduced amino acid similarity, with that of HUB1, NX06, BJsy06, VR-2332, HB-1 (sh)2002, HB-2 (sh)2002, CH-la and RespPRRS MLV respectively; and shared only 52.9%, 65.0% and 64.3% of nucleotide similarity, and 31.3%, 57.5% and 58.9% of deduced amino acid similarity with that of LV respectively. The phylogenetic tree revealed XH-GD had closer relationship with HUB1, NX06, BJsy06, VR-2332, HB-1(sh)2002, HB-2(sh)2002, CH-la and RespPRRS MLV strains than that of LV strain.

       

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