Instrument-Free Point-of-Care Molecular Detection of Zika Virus.
Instrument-Free Point-of-Care Molecular Detection of Zika Virus.
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DOI：
10.1021/acs.analchem.6b01632
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发表时间：
2016-07-19
影响因子：
7.4
通讯作者：
Liu C
中科院分区：
文献类型：
--
作者：
Song J;Mauk MG;Hackett BA;Cherry S;Bau HH;Liu C
最近在美洲爆发的寨卡病毒（ZIKV）感染及其对胎儿发育的破坏性影响促使世界卫生组织（WHO）宣布寨卡病毒大流行为国际关注的突发公共卫生事件。ZIKV的快速和可靠的诊断是至关重要的，因为ZIKV感染者没有表现出类似于其他病毒感染的症状或非特异性症状。由于免疫测定缺乏足够的灵敏度和选择性，并且无法识别感染的活跃状态，因此分子诊断是在感染后不久和整个妊娠期间检测ZIKV的有效手段。我们报告了用于快速检测ZIKV的高灵敏度逆转录环介导等温扩增（RT-LAMP）测定及其在简单、易于使用、廉价的护理点（POC）一次性盒中的实施，所述盒执行从样品引入到检测的所有单元操作。对于卡匣的热控制，我们使用化学加热杯，而不需要电力。扩增产物用无色结晶紫（LCV）染料通过眼睛检测，而不需要仪器。我们通过在不到40分钟内检测口腔样本中的ZIKV，灵敏度为5个噬菌斑形成单位（PFU），证明了我们的POC诊断系统的实用性。我们的系统特别适用于资源匮乏的环境，其中集中的实验室设施，资金和训练有素的人员短缺，并可用于医生办公室，诊所和家中。
The recent outbreak of Zika virus (ZIKV) infection in the Americas and its devastating impact on fetal development have prompted the World Health Organization (WHO) to declare the ZIKV pandemic as a Public Health Emergency of International Concern. Rapid and reliable diagnostics for ZIKV are vital because ZIKV-infected individuals display no symptoms or nonspecific symptoms similar to other viral infections. Because immunoassays lack adequate sensitivity and selectivity and are unable to identify active state of infection, molecular diagnostics are an effective means to detect ZIKV soon after infection and throughout pregnancy. We report on a highly sensitive reverse-transcription loop-mediated, isothermal amplification (RT-LAMP) assay for rapid detection of ZIKV and its implementation in a simple, easy-to-use, inexpensive, point-of-care (POC) disposable cassette that carries out all the unit operations from sample introduction to detection. For thermal control of the cassette, we use a chemically heated cup without a need for electrical power. Amplification products are detected with leuco crystal violet (LCV) dye by eye without a need for instrumentation. We demonstrated the utility of our POC diagnostic system by detecting ZIKV in oral samples with sensitivity of 5 plaque-forming units (PFU) in less than 40 min. Our system is particularly suitable for resource-poor settings, where centralized laboratory facilities, funds, and trained personnel are in short supply, and for use in doctors’ offices, clinics, and at home.