Deep sequence characterisation of a divergent HPIV-4a from an adult with prolonged influenza-like illness

Katherine E. Arden , Scott A. Beatson , Stephen B. Lambert , Claire Y.T. Wang , Jodie McVernon , Michael D. Nissen , Terry Nolan , Theo P. Sloots , Ian M. Mackay
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Abstract

Human parainfluenza virus 4 (HPIV-4) subtypes 4a and 4b are seldom sought during molecular diagnostic screening of respiratory samples from patients with influenza like illnesses (ILIs). Nonetheless, HPIV-4a and HPIV-4b are to be found in such cases, occasionally in the absence of another pathogen. Little is known about the spectrum of genetic variation among HPIV-4 genotypes; thus the impact of genetic change on transmission, pathogenicity, and shedding cannot yet be quantified. We deduced the near-complete genome of a divergent genotype of HPIV-4a (QPID08-0015) identified in a respiratory tract sample from an adult with prolonged ILI in Victoria, Australia, in 2008. Two other variants had been previously reported from Denmark during 2002–2003 (HPIV-4a|DK(459)) and Japan in 2010 (HPIV-4a| 321-Yamagata-2010).

A novel concentration, enrichment, purification and amplification (CEPA) deep sequencing process yielded > 90% coverage of the 17,140 bp HPIV-4a-QPID08-0015 genome, including all coding and intergenic regions using material from a single stored clinical sample. Genomic variation was highest between coding regions (Alquezar-Planas et al., 2013).

Deep sequencing allowed identification and genomic characterisation of a possible pathogen from an ILI as well as being an important tool to aid future understanding of the linkages between viral genetic variation, transmission and disease prognosis.

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长时间流感样疾病成人分化型HPIV-4a的深序列特征
在对流感样疾病(ILIs)患者呼吸道样本进行分子诊断筛查时,很少发现人副流感病毒4 (HPIV-4)亚型4a和4b。尽管如此,HPIV-4a和HPIV-4b在这些病例中被发现,偶尔在没有其他病原体的情况下。人们对HPIV-4基因型之间的遗传变异谱知之甚少;因此,遗传变化对传播、致病性和脱落的影响尚不能量化。我们推断出2008年在澳大利亚维多利亚州一名患有长期ILI的成人呼吸道样本中发现的HPIV-4a (QPID08-0015)的一种不同基因型的接近完整基因组。先前在2002-2003年丹麦(HPIV-4a|DK(459))和2010年日本(HPIV-4a| 321-Yamagata-2010)报告了另外两种变体。一种新的浓缩、富集、纯化和扩增(CEPA)深度测序方法获得了>HPIV-4a-QPID08-0015基因组的90%覆盖率,包括所有编码区和基因间区,使用的材料来自单个存储的临床样本。基因组变异在编码区之间最高(Alquezar-Planas et al., 2013)。深度测序可以识别和鉴定来自ILI的可能病原体的基因组特征,并且是帮助未来了解病毒遗传变异、传播和疾病预后之间联系的重要工具。
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