Development of a multiplex polymerase chain reaction assay for simultaneous identification of human enterovirus 71 and coxsackievirus A16.
Development of a multiplex polymerase chain reaction assay for simultaneous identification of human enterovirus 71 and coxsackievirus A16.
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DOI：
10.1016/j.jviromet.2010.09.017
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发表时间：
2010-12
影响因子：
3.1
通讯作者：
Phuektes, Patchara
中科院分区：
文献类型：
--
作者：
Nguyen Thi Thanh Thao;Nguyen Thi Kim Ngoc;Phan Van Tu;Tran Thi Thuy;Cardosa, Mary Jane;McMinn, Peter Charles;Phuektes, Patchara
人类肠道病毒71型（HEV 71）和柯萨奇病毒A16型（CVA 16）是引起儿童手足口病（HFMD）的两种主要病原体。最近在越南和亚太地区发生了几次大规模的手足口病疫情。本研究建立了一种同时检测HEV 71、CVA 16等肠道病毒的多重RT-PCR方法。用3对寡核苷酸引物的混合物进行肠道病毒检测：一对已发表的引物用于扩增所有已知的肠道病毒基因组，两对新的引物用于检测HEV 71和CVA 16的VP 1基因。同时对从手足口病患者中分离的肠道病毒CVA 16和HEV 71进行检测，以评价该方法的敏感性和特异性。然后将该检测方法应用于从越南胡志明市第二儿童医院鉴定的手足口病病例中获得的临床标本中直接检测这些病毒。多重RT-PCR方法在肠道病毒的筛选和鉴定HEV 71和CVA 16中显示出100%的特异性。当使用多重RT-PCR检测从医院确定的手足口病病例中获得的临床标本中筛查肠道病毒并鉴定HEV 71和CVA 16时，获得了类似的结果。该多重RT-PCR方法可用于手足口病患者HEV 71和CVA 16感染的快速、敏感和特异性诊断，并可用于分子流行病学调查。
Human enterovirus 71 (HEV71) and coxsackievirus A16 (CVA16) are two major aetiological agents of hand, foot and mouth disease (HFMD) in children. Recently there have been several large outbreaks of HFMD in Vietnam and the Asia-Pacific region. In this study, a multiplex RT-PCR assay was developed in order to detect simultaneously HEV71, CVA16 and other human enteroviruses. Enterovirus detection was performed with a mixture of three pairs of oligonucleotide primers: one pair of published primers for amplifying all known enterovirus genomes and two new primer pairs specific for detection of the VP1 genes of HEV71 and CVA16. Enterovirus isolates, CVA16 and HEV71 strains identified previously from patients with HFMD were examined to evaluate the sensitivity and specificity of the multiplex RT-PCR assay. The assay was then applied to the direct detection of these viruses in clinical specimens obtained from HFMD cases identified at Children's Hospital Number 2, Ho Chi Minh City, Vietnam. The multiplex RT-PCR assay showed 100% specificity in screening for enteroviruses and in identifying HEV71 and CVA16. Similar results were obtained when using the multiplex RT-PCR assay to screen for enteroviruses and to identify HEV71 and CVA16 in clinical specimens obtained from HFMD cases identified at the hospital. This multiplex RT-PCR assay is a rapid, sensitive and specific assay for the diagnosis of HEV71 or CVA16 infection in cases of HFMD and is also potentially useful for molecular epidemiological investigations.
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影响因子：
5.4
作者：
Oberste, MS;Maher, K;Pallansch, MA
通讯作者：
Pallansch, MA
影响因子：
8.8
作者：
Leitch, E. C. McWilliam;Harvala, H.;Simmonds, R.
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DOI：
10.1590/s0036-46651995000300009
发表时间：
1995-01-01
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作者：
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通讯作者：
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影响因子：
8.8
作者：
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通讯作者：
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影响因子：
2.6
作者：
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通讯作者：
Nishio, O