Influenza A virus isolation, culture and identification.

Influenza A virus isolation, culture and identification.
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DOI:
10.1038/nprot.2014.180
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发表时间:
2014-11
期刊:
影响因子:
14.8
通讯作者:
Kawaoka Y
Kawaoka Y
中科院分区:
生物学1区
文献类型:
--
作者:
Eisfeld AJ;Neumann G;Kawaoka Y

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甲型流感病毒(IAV)引起流行病和大流行,导致相当大的经济负担和人类生命损失。为了管理每年的IAV流行并为未来的大流行做好准备,迫切需要进一步了解IAV如何出现,传播,引起疾病和获得大流行潜力。获得这种知识所必需的基本技术是从实验和监测样品中分离和培养IAV。在这里,我们提出了一个详细的协议,用于IAV样品收集和处理,在鸡蛋和哺乳动物细胞中扩增,并从含有未知病原体的样品中进行鉴定。该方案是稳健的,并且允许生成可用于下游分析的病毒培养物。一旦获得实验或监测样本,就可以进行病毒培养,并在3-5天内验证IAV的存在。对于经验不足的实验室人员,或者需要处理大量样本时,可能需要增加时间范围。
Influenza A viruses (IAV) cause epidemics and pandemics that result in considerable financial burden and loss of human life. To manage annual IAV epidemics and prepare for future pandemics, improved understanding of how IAVs emerge, transmit, cause disease, and acquire pandemic potential is urgently needed. Fundamental techniques essential for procuring such knowledge are IAV isolation and culture from experimental and surveillance samples. Here, we present a detailed protocol for IAV sample collection and processing, amplification in chicken eggs and mammalian cells, and identification from samples containing unknown pathogens. This protocol is robust, and allows for generation of virus cultures that can be used for downstream analyses. Once experimental or surveillance samples are obtained, virus cultures can be generated and the presence of IAV can be verified in 3–5 days. Increased time-frames may be required for less experienced laboratory personnel, or when large numbers of samples will be processed.
人类感染新型禽源甲型流感 (H7N9) 病毒。
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