Reducing amplification cycles to improve the coverage of influenza A virus genome sequencing in heterosubtypic co-infection

IF 2.2 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-09-20 DOI:10.1016/j.jviromet.2024.115036
Yijie Zhang , Wenhua Kong , Yixuan Wu , Zhi Chen , Xiang Zhao , Manqing Liu
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Abstract

This study delineates the enhancement of a Reverse Transcription Polymerase Chain Reaction (RT-PCR) method for the amplification of the complete genome of the influenza A virus during heterosubtypic co-infection, relying on the amplification of intact gene segments. The precision of the method was assessed using all amplicons, which underwent both capillary electrophoresis and DNA sequencing. Five samples featuring co-infection of Influenza A viruses with H1N1 and H3N2 subtypes were evaluated. The improved strategy successfully amplified all eight segments of H3N2 strains in four samples, and the entire genome of H1N1 strains in three samples.
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减少扩增周期,提高甲型流感病毒基因组测序在异种亚型共同感染中的覆盖率。
本研究描述了一种反转录聚合酶链反应(RT-PCR)方法的改进情况,该方法可在异种亚型共感染过程中依靠完整基因片段的扩增来扩增甲型流感病毒的完整基因组。使用所有扩增子评估了该方法的精确度,这些扩增子都经过了毛细管电泳和 DNA 测序。对五份同时感染甲型 H1N1 和 H3N2 亚型流感病毒的样本进行了评估。改进策略成功扩增了四个样本中 H3N2 株的全部八个片段,以及三个样本中 H1N1 株的整个基因组。
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来源期刊
CiteScore
5.80
自引率
0.00%
发文量
209
审稿时长
41 days
期刊介绍: The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.
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