National Institute for Viral Disease Control and Prevention, China Center for Disease Control and Prevention, Key Laboratory for Medical Virology, National Health and Family Planning Commission, 155 Changbai Road, Changping District, Beijing, 102206, China.
Virol J. 2017 Sep 18;14(1):180. doi: 10.1186/s12985-017-0842-1.
Since the highly pathogenic H5N1 influenza caused thousands of deaths of wild bird in this area in 2005, Qinghai Lake in China has become a hot spot for study of the influence of avian influenza to migratory wild birds. However, the ecology and evolution of low pathogenic avian influenza virus in this region are limited. This project-based avian influenza surveillance in Qinghai lake region was initiated in year 2012.
Samples of wild bird feces and lake surface water were collected in Qinghai Lake in year 2012.Virus isolation was conducted on embryonated chicken eggs. The influenza A virus was determined by rRT-PCR. Virus sequences were acquired by deep sequencing. The phylogenetic correlation and molecular characteristics of the viruses were analyzed. The virus growth and infection features, receptor binding preference were studied, and pathogenicity in vitro as well as.
Two H13N8 subtype influenza viruses were isolated. The viruses are phylogenetically belong to Eurasian lineage. Most of the genes are associated with gull origin influenza virus except PB1 gene, which is most probably derived from Anseriformes virus. The evidence of interspecies reassortment was presented. The two viruses have limited growth capacity on MDCK and A549 cells while grow well in embryonated eggs. The dual receptor binding features of the two viruses was shown up. The low pathogenic features were determined by trypsin dependence plaque formation assay.
The two H13N8 subtype influenza viruses are highly associated with gull origin. The interspecies reassortment of H13 subtype virus among Anseriforme sand Charadriiformes wild birds emphasizes the importance of strengthening avian influenza surveillance in this region. This study is helpful to understand the ecology, evolution and transmission pattern of H13 subtype influenza virus globally.
自 2005 年高致病性 H5N1 流感导致该地区数千只野生鸟类死亡以来,中国青海湖已成为研究禽流感对候鸟影响的热点地区。然而,该地区低致病性禽流感病毒的生态学和进化情况有限。本项目于 2012 年开始在青海湖地区进行禽流感监测。
2012 年在青海湖采集野生鸟类粪便和湖水表面水样。在鸡胚上进行病毒分离。通过 rRT-PCR 确定流感病毒。通过深度测序获得病毒序列。分析病毒的系统发育相关性和分子特征。研究病毒的生长和感染特征、受体结合偏好,并进行体外致病性研究。
分离到两株 H13N8 亚型流感病毒。这些病毒在系统发育上属于欧亚谱系。除 PB1 基因外,大多数基因与海鸥源流感病毒有关,而 PB1 基因可能来源于雁形目病毒。存在种间重配的证据。两种病毒在 MDCK 和 A549 细胞上的生长能力有限,但在鸡胚上生长良好。两种病毒均表现出双重受体结合特征。通过胰蛋白酶依赖性蚀斑形成试验确定了低致病性特征。
这两株 H13N8 亚型流感病毒与海鸥高度相关。鸭科和鸻形目野生鸟类之间 H13 亚型病毒的种间重配强调了加强该地区禽流感监测的重要性。本研究有助于了解全球 H13 亚型流感病毒的生态学、进化和传播模式。