Molecular Characterization of Influenza A Viruses Detected in Children Attending Yangon Children’s Hospital, 2016

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Abstract

Sequence analysis of the influenza virus strains is important for molecular epidemiological studies and evolutional studies of influenza viruses as well as for the assessment of vaccine effectiveness. The aim of this study was to determine and characterize predominant subtype of influenza A viruses among children attending Yangon Children’s Hospital (YCH). It was a cross-sectional descriptive study conducted at YCH. Nasopharyngeal swabs were collected from 153 children who attended the hospital due to influenza-like illness (ILI) during January-December, 2016. Viral RNA was extracted by QIAamp® Viral Mini Kit. Matrix genes of influenza A and influenza B virus were detected by multiplex Reverse TranscriptionPolymerase Chain Reaction (RT-PCR). Influenza A virus matrix gene positive samples were subjected to subtyping. Predominant subtypes were subjected to sequencing and phylogenetic analysis of their HA gene and neuraminidase (NA) gene. Influenza viruses were detected in about 14% of children with ILI. Among them, 55% showed influenza A virus positive and 45% showed influenza B virus positive. Influenza A (H3N2) virus was found to be predominant among influenza A virus positive children accounting for 83.4%. There was one case (8.3%) of influenza A (H1N1) pdm09 virus and one case (8.3%) of unsubtyped influenza A virus. Phylogenetic analysis of HA and NA gene of two Myanmar strains of H3N2 subtype revealed that they belonged to clade 3C.2a1. They had 99.3-99.4% nucleotide identity with A/Hong Kong/ 4801/2014, vaccine strain of H3N2 subtype, that was contained in southern hemisphere influenza vaccine for 2016 and northern hemisphere vaccine for 2016-2017 season. This study generated information useful for the assessment of influenza outbreaks, selection of upcoming vaccine strains and further evolutionary and epidemiological studies on influenza viruses.
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2016年仰光儿童医院儿童甲型流感病毒检测的分子特征
流感病毒株的序列分析对流感病毒的分子流行病学研究和进化研究以及疫苗有效性评估具有重要意义。本研究的目的是确定和表征在仰光儿童医院(YCH)就诊的儿童中主要的甲型流感病毒亚型。这是一项在YCH进行的横断面描述性研究。收集了2016年1 - 12月因流感样疾病(ILI)就诊的153名儿童的鼻咽拭子。采用QIAamp®病毒迷你试剂盒提取病毒RNA。采用多重逆转录聚合酶链反应(RT-PCR)检测甲型流感病毒和乙型流感病毒的基质基因。甲型流感病毒基质基因阳性样本进行分型。对优势亚型的HA基因和神经氨酸酶(NA)基因进行测序和系统发育分析。约14%的ILI患儿检出流感病毒。其中甲型流感病毒阳性55%,乙型流感病毒阳性45%。甲型流感病毒阳性儿童以H3N2型流感病毒为主,占83.4%。甲型H1N1流感pdm09病毒1例(8.3%),无亚型甲型流感病毒1例(8.3%)。两株缅甸H3N2亚型的HA和NA基因的系统发育分析表明,它们属于3C.2a1支系。它们与2016年南半球流感疫苗和2016-2017年北半球流感疫苗中含有的H3N2亚型疫苗株A/Hong Kong/ 4801/2014核苷酸同源性为99.3 ~ 99.4%。这项研究为评估流感暴发、选择即将推出的疫苗株以及进一步开展流感病毒的进化和流行病学研究提供了有用的信息。
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