Direct identification of human enterovirus serotypes in cerebrospinal fluid by amplification and sequencing of the VP1 region
Direct identification of human enterovirus serotypes in cerebrospinal fluid by amplification and sequencing of the VP1 region
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DOI：
10.1016/j.jcv.2008.11.015
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发表时间：
2009-02-01
影响因子：
8.8
通讯作者：
Simmonds, R.
中科院分区：
文献类型：
--
作者：
Leitch, E. C. McWilliam;Harvala, H.;Simmonds, R.
背景：人类肠道病毒（HEV）是脑膜炎和其他神经系统疾病的主要原因。临床病例中HEV血清型的鉴定对于监测致病性更强的变异株的出现、流行病学监测和传染源调查具有重要意义。血清型鉴定是目前的问题后，广泛采用的聚合酶链反应（PCR）为基础的方法检测戊型肝炎病毒的地方病毒culture.Objectives：发展一种可靠的，灵敏的方法来确定种A和B血清型直接从脑脊液（CSF）标本。采用巢式PCR扩增HEV A、B种的VP 1区序列，与参考株进行比较，可明确地进行血清型鉴定。在19个月的研究期间收集的64份诊断性CSF样本中，有62份成功扩增（灵敏度为97%），而通过粪便和咽拭子样本的病毒培养鉴定的22份中有9份（41%）成功扩增。其中，60份样品含有种B，2份样品含有种A血清型（柯萨奇病毒A6和肠道病毒71）。观察到血清型频率和多样性的快速变化;埃可病毒（E）9型感染在2007年初占主导地位，在2007年晚些时候被E30所取代，随后在2008年被8种不同的B血清型所取代。因此，建立一种简便、快速的方法，直接从脑脊液中鉴定血清型和单个HEV株或群，将具有重要意义在监测方面的价值，更多地了解与疾病类型相关的差异，并监测肠道病毒71型等致病性变体的全球传播。(C)2008 Elsevier B. V.保留所有权利。
Background: Human enteroviruses (HEV) are a major cause of meningitis and other neurological disease. Identification of HEV serotypes in clinical cases is important for monitoring emergence of more pathogenic variants, epidemiological surveillance and investigating sources of infection. Serotype identification is currently problematic following the widespread adoption of polymerase chain reaction (PCR)-based methods for HEV detection in place of virus culture.Objectives: To develop a reliable, sensitive method to identify species A and B serotypes directly from cerebrospinal fluid (CSF) specimens.Study design: A nested-PCR was used to amplify VP1 region sequences of HEV species A and B, that enabled unambiguous serotype identification by comparison with reference strains.Results: 62 from 64 diagnostic CSF samples collected over a 19-month study period were successfully amplified (97% sensitivity), compared with 9/22 (41%) identified by virus culture of co-referred faecal and throat swab samples. Among these, 60 samples contained species B and 2 samples contained species A serotypes (coxsackievirus A6 and enterovirus 71) were identified. Rapid changes in serotype frequencies and diversity were observed; echovirus (E) type 9 infections predominated in early 2007, to be replaced by E30 later in the year and followed by a diverse range of eight different species B serotypes in 2008.Conclusions: The availability of a simple and rapid method for identification of serotypes and individual HEV strains or clusters directly from CSF will be of substantial value in surveillance, understanding more about serotype-associated differences in disease and monitoring the global spread of pathogenic variants such as enterovirus 71. (C) 2008 Elsevier B.V. All rights reserved.