Simultaneous detection of severe acute respiratory syndrome, Middle East respiratory syndrome, and related bat coronaviruses by real-time reverse transcription PCR.
Simultaneous detection of severe acute respiratory syndrome, Middle East respiratory syndrome, and related bat coronaviruses by real-time reverse transcription PCR.
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DOI：
10.1007/s00705-017-3281-9
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发表时间：
2017-06
影响因子：
2.7
通讯作者：
Kim HK
中科院分区：
文献类型：
--
作者：
Noh JY;Yoon SW;Kim DJ;Lee MS;Kim JH;Na W;Song D;Jeong DG;Kim HK
由于严重急性呼吸综合征(SARS)和中东呼吸综合征(MERS)冠状病毒(CoV)在临床症状、病原学和传播方面具有相似的特征，因此快速准确的鉴别诊断方法非常重要。因此，本研究的目的是建立一种同时检测这些病毒的双重实时逆转录-聚合酶链式反应(RT-PCR)方法。设计了针对人SARS冠状病毒、MERS冠状病毒及其相关蝙蝠冠状病毒保守的尖峰S2区的引物和探针。实时RT-PCR结果显示，分别使用pUC57-SARS-PS2(SARS-CoV模板)和pGEM-MERS-S2(MERS-CoV模板)对每种病毒进行了特异性反应，检测下限分别为50-100和5-100拷贝/毫升。此外，该实时荧光RT-PCR系统还能在MERS患者的BAT粪便样本和痰标本中分别检测到类似SARS的BAT冠状病毒和MERS冠状病毒。因此，这一新开发的实时荧光RT-PCR方法不仅有望在人类中检测到SARS-CoV和MERS-CoV，还有望在蝙蝠中检测到与这些病毒密切相关的几种蝙蝠冠状病毒。
Since severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronaviruses (CoVs) share similar characteristics with respect to clinical signs, etiology, and transmission, methods for a rapid and accurate differential diagnosis are important. Therefore, the aim of this study was to develop a duplex real-time reverse transcription (RT)-PCR method for the simultaneous detection of these viruses. Primers and probes that target the conserved spike S2 region of human SARS-CoV, MERS-CoV, and their related bat CoVs were designed. The results of real-time RT-PCR showed specific reactions for each virus with adequate detection limits of 50–100 copies/mL and 5–100 copies/mL using pUC57-SARS-pS2 (a template for SARS-CoV) and pGEM-MERS-S2 (a template for MERS-CoV), respectively. In addition, this real-time RT-PCR system was able to detect the target viruses SARS-like bat CoV and MERS-CoV in bat fecal samples and sputum of MERS patients, respectively. Therefore, this newly developed real-time RT-PCR method is expected to detect not only SARS-CoV and MERS-CoV in humans but also several bat CoVs that are closely related to these viruses in bats.
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影响因子：
82.9
作者：
Menachery VD;Yount BL Jr;Debbink K;Agnihothram S;Gralinski LE;Plante JA;Graham RL;Scobey T;Ge XY;Donaldson EF;Randell SH;Lanzavecchia A;Marasco WA;Shi ZL;Baric RS
通讯作者：
Baric RS
DOI：
10.1038/nrmicro.2016.81
发表时间：
2016-08
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
de Wit E;van Doremalen N;Falzarano D;Munster VJ
通讯作者：
Munster VJ
影响因子：
9.8
作者：
Chowell G;Nishiura H
通讯作者：
Nishiura H
影响因子：
5.4
作者：
Yang, Xing-Lou;Hu, Ben;Shi, Zheng-Li
通讯作者：
Shi, Zheng-Li
影响因子：
2.7
作者：
Hadjinicolaou, Andreas V.;Farcas, Gabriella A.;Kostrikis, Leondios G.
通讯作者：
Kostrikis, Leondios G.