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利用合成 DNA 快速生成大流行性流感病毒疫苗候选株。

Rapid generation of pandemic influenza virus vaccine candidate strains using synthetic DNA.

机构信息

CSL Ltd, Parkville, Vic., Australia.

出版信息

Influenza Other Respir Viruses. 2012 Mar;6(2):101-9. doi: 10.1111/j.1750-2659.2011.00273.x. Epub 2011 Jul 19.

DOI:10.1111/j.1750-2659.2011.00273.x
PMID:21771285
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4942080/
Abstract

BACKGROUND

Vaccination is considered the most effective means of reducing influenza burden. The emergence of H5N1 and pandemic spread of novel H1N1/2009 viruses reinforces the need to have strategies in place to rapidly develop seed viruses for vaccine manufacture.

METHODS

Candidate pandemic vaccine strains consisting of the circulating strain haemagglutinin (HA) and neuraminidase (NA) in an A/PR/8/34 backbone were generated using alternative synthetic DNA approaches, including site-directed mutagenesis of DNA encoding related virus strains, and rapid generation of virus using synthetic DNA cloned into plasmid vectors.

RESULTS

Firstly, synthetic A/Bar Headed Goose/Qinghai/1A/2005 (H5N1) virus was generated from an A/Vietnam/1194/2004 template using site-directed mutagenesis. Secondly, A/Whooper Swan/Mongolia/244/2005 (H5N1) and A/California/04/09 (H1N1) viruses were generated using synthetic DNA encoding the viral HA and NA genes. Replication and antigenicity of the synthetic viruses were comparable to that of the corresponding non-synthetic viruses.

CONCLUSIONS

In the event of an influenza pandemic, the use of these approaches may significantly reduce the time required to generate and distribute the vaccine seed virus and vaccine manufacture. These approaches also offer the advantage of not needing to handle wild-type virus, potentially diminishing biocontainment requirements.

摘要

背景

接种疫苗被认为是减轻流感负担的最有效手段。H5N1 的出现和新型 H1N1/2009 病毒的大流行传播，强化了需要制定策略，以便快速开发用于疫苗生产的种子病毒。

方法

候选大流行疫苗株由 A/PR/8/34 骨架中的流行株血凝素 (HA) 和神经氨酸酶 (NA) 组成，采用替代的合成 DNA 方法生成，包括对相关病毒株的 DNA 进行定点诱变，以及使用合成 DNA 克隆到质粒载体中快速生成病毒。

结果

首先，使用定点诱变从 A/Vietnam/1194/2004 模板生成了合成的 A/Bar Headed Goose/Qinghai/1A/2005 (H5N1) 病毒。其次，使用编码病毒 HA 和 NA 基因的合成 DNA 生成了 A/Whooper Swan/Mongolia/244/2005 (H5N1) 和 A/California/04/09 (H1N1) 病毒。合成病毒的复制和抗原性与相应的非合成病毒相当。

结论

在流感大流行的情况下，这些方法的使用可能会显著缩短生成和分发疫苗种子病毒以及疫苗生产所需的时间。这些方法还具有不需要处理野生型病毒的优势，从而降低生物安全要求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f54d/4942080/fdef21fd0550/IRV-6-101-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f54d/4942080/ad9be7e171e5/IRV-6-101-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f54d/4942080/fdef21fd0550/IRV-6-101-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f54d/4942080/ad9be7e171e5/IRV-6-101-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f54d/4942080/fdef21fd0550/IRV-6-101-g002.jpg

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