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利用完整血凝素-神经氨酸酶(HN)基因对肯尼亚分离的人类呼吸道合胞病毒 3 型毒株的遗传多样性和进化分析。

Genetic diversity and evolutionary analysis of human respirovirus type 3 strains isolated in Kenya using complete hemagglutinin-neuraminidase (HN) gene.

机构信息

Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, Juja, Kenya.

Department of Emerging infections, US Army Medical Directorate-Africa, Nairobi, Kenya.

出版信息

PLoS One. 2020 Mar 10;15(3):e0229355. doi: 10.1371/journal.pone.0229355. eCollection 2020.

Abstract

Human respirovirus type 3 (HRV3) is a leading etiology of lower respiratory tract infections in young children and ranks only second to the human respiratory syncytial virus (HRSV). Despite the public health importance of HRV3, there is limited information about the genetic characteristics and diversity of these viruses in Kenya. To begin to address this gap, we analyzed 35 complete hemagglutinin-neuraminidase (HN) sequences of HRV3 strains isolated in Kenya between 2010 and 2013. Viral RNA was extracted from the isolates, and the entire HN gene amplified by RT-PCR followed by nucleotide sequencing. Phylogenetic analyses of the sequences revealed that all the Kenyan isolates grouped into genetic Cluster C; sub-clusters C1a, C2, and C3a. The majority (54%) of isolates belonged to sub-cluster C3a, followed by C2 (43%) and C1a (2.9%). Sequence analysis revealed high identities between the Kenyan isolates and the HRV3 prototype strain both at the amino acid (96.5-97.9%) and nucleotide (94.3-95.6%) levels. No amino acid variations affecting the catalytic/active sites of the HN glycoprotein were observed among the Kenyan isolates. Selection pressure analyses showed that the HN glycoprotein was evolving under positive selection. Evolutionary analyses revealed that the mean TMRCA for the HN sequence dataset was 1942 (95% HPD: 1928-1957), while the mean evolutionary rate was 4.65x10-4 nucleotide substitutions/site/year (95% HPD: 2.99x10-4 to 6.35x10-4). Overall, our results demonstrate the co-circulation of strains of cluster C HRV3 variants in Kenya during the study period. This is the first study to describe the genetic and molecular evolutionary aspects of HRV3 in Kenya using the complete HN gene.

摘要

人类呼肠孤病毒 3 型(HRV3)是导致婴幼儿下呼吸道感染的主要病原体,其地位仅次于人类呼吸道合胞病毒(HRSV)。尽管 HRV3 具有重要的公共卫生意义,但关于肯尼亚这些病毒的遗传特征和多样性的信息有限。为了开始解决这一差距,我们分析了 2010 年至 2013 年间在肯尼亚分离的 35 株 HRV3 完整血凝素-神经氨酸酶(HN)序列。从分离株中提取病毒 RNA,通过 RT-PCR 扩增全长 HN 基因,然后进行核苷酸测序。序列的系统进化分析表明,所有肯尼亚分离株均属于遗传簇 C;亚群 C1a、C2 和 C3a。大多数(54%)分离株属于 C3a 亚群,其次是 C2(43%)和 C1a(2.9%)。序列分析显示,肯尼亚分离株与 HRV3 原型株在氨基酸(96.5-97.9%)和核苷酸(94.3-95.6%)水平上具有高度同源性。在肯尼亚分离株中没有观察到影响 HN 糖蛋白催化/活性位点的氨基酸变异。HN 糖蛋白在正选择下进化。进化分析显示,HN 序列数据集的 TMRCA 平均值为 1942(95%HPD:1928-1957),而平均进化率为 4.65x10-4 核苷酸替换/位点/年(95%HPD:2.99x10-4 至 6.35x10-4)。总的来说,我们的研究结果表明,在研究期间,肯尼亚存在 C 簇 HRV3 变体的流行株。这是首次使用完整的 HN 基因描述肯尼亚 HRV3 的遗传和分子进化方面的研究。

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