RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19
RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19
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DOI：
10.1038/s41596-021-00546-w
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发表时间：
2021-04-30
期刊：
影响因子：
14.8
通讯作者：
Pan, Dipanjan
中科院分区：
文献类型：
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作者：
Alafeef, Maha;Moitra, Parikshit;Pan, Dipanjan
该方案提供了一种无需RNA提取的纳米放大比色测试，可以用肉眼快速检测SARS-CoV-2。该测试使用带有反义寡核苷酸的等离子体金纳米颗粒作为比色生物传感器，用于COVID-19的床旁诊断。2019年冠状病毒病（COVID-19）全球大流行凸显了当前病毒性疾病诊断测试范式的缺点。在这里，我们报告了一种用于COVID-19快速和裸眼分子诊断的无RNA提取纳米放大比色测试的分步方案。该检测采用独特的双管齐下的方法，将核酸（NA）扩增和等离子体传感集成在一起，用于对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的即时检测，样品到检测响应时间分别为98.4%，>96.6%和100%，检测限为10拷贝/μ L。该测试可以很容易地适用于诊断其他病毒感染与一个简单的修改的ASO和引物序列。它还提供了一种低成本、快速的方法，需要最少的仪器，可用作在资源匮乏的情况下在护理点诊断COVID-19的筛查工具。测试的比色读数甚至可以使用手持式光学读取器进行监测，以获得定量响应。因此，我们预计该方案将广泛用于开发用于COVID-19和其他传染病分子诊断的生物传感器。
This protocol provides an RNA extraction-free nano-amplified colorimetric test that enables rapid detection of SARS-CoV-2 with the naked eye. The test uses plasmonic gold nanoparticles capped with antisense oligonucleotides as a colorimetric biosensor for point-of-care diagnosis of COVID-19.The global pandemic of coronavirus disease 2019 (COVID-19) highlights the shortcomings of the current testing paradigm for viral disease diagnostics. Here, we report a stepwise protocol for an RNA-extraction-free nano-amplified colorimetric test for rapid and naked-eye molecular diagnosis of COVID-19. The test employs a unique dual-prong approach that integrates nucleic acid (NA) amplification and plasmonic sensing for point-of-care detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with a sample-to-assay response time of 98.4%, >96.6% and 100%, respectively, with a detection limit of 10 copies/mu L. The test can easily be adapted to diagnose other viral infections with a simple modification of the ASOs and primer sequences. It also provides a low-cost, rapid approach requiring minimal instrumentation that can be used as a screening tool for the diagnosis of COVID-19 at point-of-care settings in resource-poor situations. The colorimetric readout of the test can even be monitored using a handheld optical reader to obtain a quantitative response. Therefore, we anticipate that this protocol will be widely useful for the development of biosensors for the molecular diagnostics of COVID-19 and other infectious diseases.