Survey and Visual Detection of Zaire ebolavirus in Clinical Samples Targeting the Nucleoprotein Gene in Sierra Leone.
Survey and Visual Detection of Zaire ebolavirus in Clinical Samples Targeting the Nucleoprotein Gene in Sierra Leone.
复制标题
塞拉利昂临床样本中针对核蛋白基因的扎伊尔埃博拉病毒的调查和目视检测。
DOI：
10.3389/fmicb.2015.01332
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发表时间：
2015
影响因子：
5.2
通讯作者：
Yuan J
中科院分区：
文献类型：
--
作者：
Li H;Wang X;Liu W;Wei X;Lin W;Li E;Li P;Dong D;Cui L;Hu X;Li B;Ma Y;Zhao X;Liu C;Yuan J
埃博拉病毒（EBOV）可导致严重的出血热，人类和其他灵长类动物死亡的风险很高。为了指导治疗并防止病毒感染的传播，临床样本需要一种快速、灵敏的检测方法。在这里，我们描述并评估了一种使用核蛋白基因（NP）作为靶序列检测扎伊尔埃博拉病毒的逆转录环介导等温扩增（RT-LAMP）方法。使用了两种不同的技术：钙黄绿素/Mn2+复合显色法和实时浊度监测。 RT-LAMP 检测在 61°C 下 45 分钟内检测到 NP 靶序列，限制为 4.56 拷贝/μL，与实时逆转录聚合酶链式反应 (RT-PCR) 的灵敏度相似或有所提高。此外，所有假病毒颗粒或非扎伊尔 EBOV 基因组的 LAMP 检测均为阴性，表明该检测对 EBOV 具有高度特异性。为了评估 RT-LAMP 方法用于 EBOV 临床诊断的可用性，从塞拉利昂临床疑似感染患者收集的 417 份血液或拭子样本中，确定了 307 份用于基于 RT-LAMP 的 EBOV 监测。因此，高度特异性和敏感性的RT-LAMP方法可以快速检测埃博拉病毒，是临床筛查、诊断和初级检疫目的的合适工具。
Ebola virus (EBOV) can lead to severe hemorrhagic fever with a high risk of death in humans and other primates. To guide treatment and prevent spread of the viral infection, a rapid and sensitive detection method is required for clinical samples. Here, we described and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect Zaire ebolavirus using the nucleoprotein gene (NP) as a target sequence. Two different techniques were used, a calcein/Mn2+ complex chromogenic method and real-time turbidity monitoring. The RT-LAMP assay detected the NP target sequence with a limit of 4.56 copies/μL within 45 min under 61°C, a similar even or increase in sensitivity than that of real-time reverse transcription-polymerase chain reaction (RT-PCR). Additionally, all pseudoviral particles or non- Zaire EBOV genomes were negative for LAMP detection, indicating that the assay was highly specific for EBOV. To appraise the availability of the RT-LAMP method for use in clinical diagnosis of EBOV, of 417 blood or swab samples collected from patients with clinically suspected infections in Sierra Leone, 307 were identified for RT-LAMP-based surveillance of EBOV. Therefore, the highly specific and sensitive RT-LAMP method allows the rapid detection of EBOV, and is a suitable tool for clinical screening, diagnosis, and primary quarantine purposes.