Hangzhou Center for Disease Control and Prevention, Zhejiang, China.
J Virol. 2014 Mar;88(6):3423-31. doi: 10.1128/JVI.02059-13. Epub 2014 Jan 8.
Avian influenza virus A of the novel H7N9 reassortant subtype was recently found to cause severe human respiratory infections in China. Live poultry markets were suspected locations of the human H7N9 infection sources, based on the cases' exposure histories and sequence similarities between viral isolates. To explore the role of live poultry markets in the origin of the novel H7N9 virus, we systematically examined poultry and environmental specimens from local markets and farms in Hangzhou, using real-time reverse transcription-PCR (RT-PCR) as well as high-throughput next-generation sequencing (NGS). RT-PCR identified specimens positive for the H7 and N9 genomic segments in all of the 12 poultry markets epidemiologically linked to 10 human H7N9 cases. Chickens, ducks, and environmental specimens from the markets contained heavily mixed subtypes, including H7, N9, H9, and N2 and sometimes H5 and N1. The idea of the coexistence of H7N9 and H9N2 subtypes in chickens was further supported by metagenomic sequencing. In contrast, human H7N9 infection cases (n = 31) were all negative for H9N2 virus according to real-time RT-PCR. The six internal segments were indistinguishable for the H7N9 and H9N2 viruses. The H9, N2, and internal-segment sequences were very close to the sequence of the H9N2 virus circulating in chickens in China recently. Our results provide direct evidence that H9N2 strains coexisted with the novel human-pathogenic H7N9 influenza virus in epidemiologically linked live poultry markets. Avian influenza A virus of the H9N2 subtype likely made a recent contribution to the evolution of the H7N9 virus and continues to do so.
Our results suggest that avian influenza A virus of the H9N2 subtype likely made a recent contribution to the evolution of the H7N9 virus, a novel reassortant avian influenza virus A subtype, and continues to do so. The finding helps shed light on how the H7N9 virus emerged, spread, and transmitted to humans. It is of considerable interest for assessing the risk of the possible emergence of novel reassortant viruses with enhanced transmissibility to humans.
本研究旨在探索活禽市场在新型 H7N9 病毒起源中的作用,我们系统地检测了来自杭州当地市场和农场的家禽和环境样本,采用实时 RT-PCR 和高通量下一代测序(NGS)。
对与 10 例人感染 H7N9 病例相关的 12 个农贸市场进行流行病学调查,采用实时 RT-PCR 检测 H7 和 N9 基因组片段,采用高通量 NGS 检测 H7、N9、H9 和 N2 等混合亚型。
在所有与 10 例人感染 H7N9 病例相关的 12 个农贸市场中,均从流行病学上相关的家禽和环境标本中检测到 H7 和 N9 基因组片段。鸡、鸭和市场环境标本中存在 H7、N9、H9 和 N2 等混合亚型,有时还存在 H5 和 N1 亚型。宏基因组测序进一步支持了 H7N9 和 H9N2 亚型在鸡中共同存在的观点。相反,根据实时 RT-PCR,31 例人感染 H7N9 病例均未检测到 H9N2 病毒。H7N9 和 H9N2 病毒的 6 个内部片段完全相同。H9、N2 和内部片段序列与中国近期鸡中流行的 H9N2 病毒序列非常接近。
我们的研究结果为 H9N2 株与新型人致病性 H7N9 流感病毒在流行病学相关的活禽市场中共同存在提供了直接证据。H9N2 亚型禽流感病毒可能对 H7N9 病毒的进化做出了近期贡献,并将继续发挥作用。