Whole genome sequence analysis of cell culture-adapted rotavirus A strains from chicken.
Whole genome sequence analysis of cell culture-adapted rotavirus A strains from chicken.
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鸡细胞培养适应性轮状病毒 A 株的全基因组序列分析
DOI：
10.1016/j.meegid.2020.104275
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Tausch
中科院分区：
文献类型：
--
作者：
Patzina-Mehling;Falkenhagen;Gadicherla;Grützke;Tausch
轮状病毒A（RVA）是人类和哺乳动物胃肠炎的主要原因，并且在禽类物种中也被大量检测到。禽RVA感染与腹泻、生长减缓和死亡率增加有关，导致家禽业的经济损失。鸟类RVA在整个RVA物种中形成独特的遗传分支。然而，迄今为止，只有少数的禽RVA全基因组被公开，也只有少数的禽RVA株被适应细胞培养用于后续的研究。来自培养上清液的病毒的透射电子显微镜显示出典型的三层形态的轮状病毒颗粒;此外，菌株06 V0661 G1显示出高比例的双层颗粒。使用下一代测序（NGS）确定了病毒的（几乎）完整基因组序列。将所得序列与先前使用桑格测序确定的菌株的全长或部分序列进行比较;并鉴定了一些核苷酸错配，其中一些导致氨基酸取代。菌株02 V0002 G3、04 V0027 G6和05 V0500 F6的基因组彼此密切相关，显示G19-P[30]-I11-R6-C6-M7-A16-N6-T8-E10-H8基因型星座。菌株06 V0661 G1在与其他菌株相同的遗传骨架中携带VP 4基因型P[31]。进一步的序列分析表明，该毒株的基因，尤其是编码NSP 3的基因，在系统发育树中与其他毒株的基因更加分离，这为进一步研究禽轮状病毒的遗传多样性和复制，以及疫苗和诊断工具的开发提供了依据。
Rotavirus A (RVA) is a major cause of gastroenteritis in humans and mammalian animals, and has also been abundantly detected in avian species. Avian RVA infection is associated with diarrhea, reduced growth and increased mortality, leading to economic losses in the poultry industry. Avian RVA forms a unique genetic clade within the whole RVA species. However, up to now, only a few full-length avian RVA genomes have been published and only a small number of avian RVA strains have been adapted to grow in cell culture for subsequent studies.Here, the four cell culture-adapted chicken RVA strains 02V0002G3, 04V0027G6, 05V0500F6 and 06V0661G1 were characterized in more detail. Transmission electron microscopy of the viruses derived from culture supernatant showed a typical triple-layered morphology of rotavirus particles; in addition, strain 06V0661G1 showed a high proportion of double-layered particles. The (nearly) complete genome sequences of the viruses were determined using next-generation sequencing (NGS). The resulting sequences were compared to full-length or partial sequences of the strains previously determined using Sanger sequencing; and a few nucleotide mismatches, some of them resulting in amino acid substitutions, were identified. The genomes of strains 02V0002G3, 04V0027G6 and 05V0500F6 were closely related to each other showing a G19-P[30]-I11-R6-C6-M7-A16-N6-T8-E10-H8 genotype constellation. Strain 06V0661G1 carries the VP4 genotype P[31] in the same genetic backbone like the other strains. However, further sequence analysis showed that the genes of this strain, especially that encoding NSP3, clustered more separately from the other strains in phylogenetic trees.The characterized cell culture-adapted chicken RVA strains may be useful for future studies investigating genetic diversity and replication of avian rotaviruses, as well as for the development of vaccines and diagnostic tools.
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