Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19)
Development of a Laboratory-safe and Low-cost Detection Protocol for SARS-CoV-2 of the Coronavirus Disease 2019 (COVID-19)
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DOI：
10.5607/en2009
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发表时间：
2020-04-01
影响因子：
2.4
通讯作者：
Lee, C. Justin
中科院分区：
文献类型：
--
作者：
Won, Joungha;Lee, Solji;Lee, C. Justin
2019年12月在中国武汉出现的严重急性呼吸道冠状病毒2 （SARS-CoV-2）已迅速传播到十几个国家。特别是，韩国病例数量的激增引发了对大流行的担忧。据报告，这种病毒主要通过咳嗽和打喷嚏产生的呼吸道飞沫进行人际接触传播，或者可能通过被咳嗽或打喷嚏的人污染的表面传播。更重要的是，有报道称，这种病毒有可能在携带病毒的人出现症状之前传播。因此，迫切需要一种低成本、易于获取的方案来早期发现这种病毒。在这里，我们建立了一种基于实时反转录PCR （rtPCR）的检测方案，包括通过咽拭子自采集样本、trizol基RNA纯化和SYBR green基rtPCR。当我们用已知的慢病毒进行测试时，该方案显示出1-10个病毒颗粒的准确性和灵敏度限制。每个样品的成本估计在15美元以下。整个方案所需的总时间估计不超过4小时。我们提出了一种在任何配备rt - pcr机的传统生物安全二级实验室中进行SARS-CoV-2早期检测的成本效益高、快速简便的方法。我们的新方案将有助于对无症状病毒携带者进行第一手筛查，进一步预防传播，并对快速传播的病毒进行早期干预和治疗。
The severe acute respiratory coronavirus 2 (SARS-CoV-2), which emerged in December 2019 in Wuhan, China, has spread rapidly to over a dozen countries. Especially, the spike of case numbers in South Korea sparks pandemic worries. This virus is reported to spread mainly through person-to-person contact via respiratory droplets generated by coughing and sneezing, or possibly through surface contaminated by people coughing or sneezing on them. More critically, there have been reports about the possibility of this virus to transmit even before a virus-carrying person to show symptoms. Therefore, a low-cost, easy-access protocol for early detection of this virus is desperately needed. Here, we have established a real-time reverse-transcription PCR (rtPCR)-based assay protocol composed of easy specimen self-collection from a subject via pharyngeal swab, Trizol-based RNA purification, and SYBR Green-based rtPCR. This protocol shows an accuracy and sensitivity limit of 1-10 virus particles as we tested with a known lentivirus. The cost for each sample is estimated to be less than 15 US dollars. Overall time it takes for an entire protocol is estimated to be less than 4 hours. We propose a cost-effective, quick-and-easy method for early detection of SARS-CoV-2 at any conventional Biosafety Level II laboratories that are equipped with a rtPCR machine. Our newly developed protocol should be helpful for a first-hand screening of the asymptomatic virus-carriers for further prevention of transmission and early intervention and treatment for the rapidly propagating virus.