2014/ 2015日本流感季节流感患者分离的A/H3N2和B型流感病毒神经氨酸酶氨基酸序列

Hideyuki Ikematsu, Yong Chong, Kenjiro Shirane, Hidehiro Toh, Hiroyuki Sasaki, Shinya Matsumoto, Nozomi Noda, Taeko Hotta, Takeshi Uchiumi, Dongchon Kang
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引用次数: 0

摘要

背景:神经氨酸酶(NA)是流感病毒复制所必需的一种表面蛋白。在日本,NA抑制剂通常用于治疗流感患者。已经报道了几种降低NA抑制剂作用的突变。我们对流感患者分离病毒的NA片段进行了全序列测序,研究了NA氨基酸序列与4种NA抑制剂50%抑制浓度(IC_50)的关系。材料与方法:对2014/15流感季节患者分离的40株A/H3N2和19株B型流感病毒进行分析。IC_50采用荧光底物神经氨酸酶抑制法测定。用RT-PCR扩增病毒RNA,用下一代测序仪对基因组进行测序。对推导出的氨基酸序列进行分析。结果:2014- 2015年流感季节分离的A/H3N2和B病毒NA催化位点无AA变化。A/H3N2和B病毒4种NA抑制剂的IC_50与NA氨基酸无显著相关性。结论:A/H3N2和B病毒NA的催化位点高度保守。未观察到与目前使用的四种NA抑制剂的敏感性相关的NA氨基酸突变的出现。
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Neuraminidase Amino Acid Sequences of Influenza A/H3N2 and B Viruses Isolated from Influenza Patients in the 2014/15 Japanese Influenza Season.

Background: Neuraminidase (NA) is a surface protein essential for influenza virus replication. NA inhibitors are commonly used for the treatment of influenza patients in Japan. Several mutations that reduce the effect of NA inhibitors have been reported. We sequenced the whole NA segment of isolated virus from influenza patients and investigated the relation between the NA amino acid sequence and the 50% inhibitory concentration (IC_50) of four NA inhibitors.

Materials and methods: Forty A/H3N2 and 19 B influenza virus isolated from patients in the 2014/15 influenza season were analyzed. The IC_50 was determined by a neuraminidase inhibition assay using a fluorescent substrate. Viral RNA was amplified by RT-PCR and the genome was sequenced using a next generation sequencer. The deduced amino acid sequences were analyzed.

Results: There was no AA change in the NA catalytic site of the A/H3N2 and B viruses isolated in the 2014-15 influenza season. There was no significant relation between the NA amino acids and the IC_50 of the four NA inhibitors for A/H3N2 or B viruses.

Conclusion: The catalytic site of NA was highly conserved for these A/H3N2 and B viruses. No emergence of NA amino acid mutations related to the sensitivity of the four currently used NA inhibitors was observed.

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