中国分离的猪流行性腹泻病毒 HLJBY 的特性分析及其进化研究。
Characterization and evolution of the coronavirus porcine epidemic diarrhoea virus HLJBY isolated in China.
机构信息
Institutes of Agricultural Science and Technology Development, College of Veterinary Medicine, Yangzhou University, Yangzhou, China.
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China.
出版信息
Transbound Emerg Dis. 2020 Jan;67(1):65-79. doi: 10.1111/tbed.13321. Epub 2019 Aug 22.
A strain of porcine epidemic diarrhoea virus (PEDV), namely HLJBY, was isolated in Heilongjiang province, China. To provide insight into the understanding of the phylogenetic and the current epidemiological status of PEDV, PEDV HLJBY was compared with CV777 and other PEDV strains deposited in the GenBank. The homology between the entire genomic nucleotide sequences of PEDV HLJBY and CV777 was 97.7%. The homology of M gene was the highest (99.0%). However, the homology of ORF3 gene was 97.7%, and protein of ORF3 was 90.1%. In addition, HLJBY showed the highest nucleotide identity (99.9%) with PEDV-SX/China/2017 strain and lowest similarity (91.2%) to PEDV/Belgorod/dom/2008 strain. We analysed the changes in S gene and its protein of PEDV HLJBY with 65 historic PEDV strains. The highest nucleotide identity was 99.9% compared with PEDV-SX/China/2017 strain, and the lowest nucleotide identity was 60.0% compared with PEDV/Belgorod/dom/2008 strain. The length of deduced amino acid sequences of S proteins varied from 1,372 to 1,390 amino acids (aa). Compared with most aa sequences of S proteins, HLJBY exhibited 5 aa deletions (position 55, 59-61, 144). Analysis and comparison of open reading frame 3 (ORF3) proteins between HLJBY strain and other PEDV strains were also focused in this study. We revealed that the length of deduced amino acid sequences of ORF3 proteins was 80-224 aa among tested strains and the identity of HLJBY ORF3 amino acids with other PEDV strains was 71.4%-98.9%. ORF3 protein of both HLJBY strain and PEDV-SX/China/2017 strain consists of 91 aa, with 133 aa deletions at their C' end in relation to the other tested PEDV strains. The phylogenetic tree based on different proteins or genes resulted in different phylogenetic groups. For pathogenicity evaluation of PEDV HLJBY strain, colostrum deprivation piglets were challenged with PEDV HLJBY, and PEDV reference strain CV777 as a control, the results showed that animals challenged with either of these PEDV strains developed diarrhoea, and histopathological examination of small intestines of challenged animals showed acute viral enteritis with villous atrophy in either PEDV HLJBY-P10 or PEDV CV777-P8 inoculated piglets.
一株猪流行性腹泻病毒（PEDV），即 HLJBY，在中国黑龙江省被分离出来。为了深入了解 PEDV 的系统发育和当前流行情况，将 PEDV HLJBY 与 CV777 及其他保存在 GenBank 中的 PEDV 株进行比较。PEDV HLJBY 与 CV777 的全基因组核苷酸序列同源性为 97.7%。M 基因的同源性最高（99.0%）。然而，ORF3 基因的同源性为 97.7%，其蛋白同源性为 90.1%。此外，HLJBY 与 PEDV-SX/中国/2017 株的核苷酸同一性最高（99.9%），与 PEDV/Belgorod/dom/2008 株的相似性最低（91.2%）。我们分析了与 65 株历史 PEDV 株的 S 基因及其蛋白的变化。与 PEDV-SX/中国/2017 株相比，其核苷酸同一性最高为 99.9%，与 PEDV/Belgorod/dom/2008 株相比，核苷酸同一性最低为 60.0%。S 蛋白推导的氨基酸序列长度从 1372 到 1390 个氨基酸（aa）不等。与大多数 S 蛋白的 aa 序列相比，HLJBY 显示 5 个 aa 缺失（位置 55、59-61、144）。本研究还重点分析和比较了 HLJBY 株与其他 PEDV 株的开放阅读框 3（ORF3）蛋白。我们揭示了测试株之间 ORF3 蛋白推导的氨基酸序列长度为 80-224aa，HLJBY ORF3 氨基酸与其他 PEDV 株的同一性为 71.4%-98.9%。HLJBY 株和 PEDV-SX/中国/2017 株的 ORF3 蛋白均由 91 个氨基酸组成，与其他测试的 PEDV 株相比，其 C'末端有 133 个 aa 缺失。基于不同蛋白质或基因的系统发育树导致不同的系统发育群。为了评估 PEDV HLJBY 株的致病性，用 PEDV HLJBY 对初乳缺乏的仔猪进行攻毒，并用 PEDV 参考株 CV777 作为对照，结果表明，用这些 PEDV 株中的任何一株攻毒的动物均出现腹泻，攻毒仔猪小肠的组织病理学检查显示，PEDV HLJBY-P10 或 PEDV CV777-P8 接种仔猪均出现急性病毒性肠炎，绒毛萎缩。
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