Lu Xiao, Lin Haosi, Feng Xianzhen, Lui Grace Cy, Hsing I-Ming
Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
Department of Medicine & Therapeutics, The Chinese University of Hong Kong, Hong Kong, China.
Biosens Bioelectron X. 2022 Dec;12:100248. doi: 10.1016/j.biosx.2022.100248. Epub 2022 Sep 5.
Rapid at-home test is a good alternative to the gold standard quantitative polymerase chain reaction (qPCR) for early identification and management of infected individuals in pandemic. However, the currently available at-home rapid antigen kits and nucleic acid tests (NATs) are prone to false results. Although some CRISPR-mediated NATs enhanced accuracy, long turnaround time (ca. 1 h) and aerosol contamination due to additional open-lid reaction hinder its applicability for self-tests. Moreover, the accuracy of at-home NATs is also impacted by interference of sample matrix due to lack of sample purification. Here we report a Fast, Low-cost, Aerosol contamination-free and Sensitive molecular assay for at-Home tests of communicable pathogens (FLASH) incorporating oLAMP, a recently reported isothermal and target-specific NATs by our group, and a visible lateral-flow readout. The integrated platform enabled sample-to-result SARS-CoV-2 RNA detection in 20-30 min achieving a sensitivity of 0.5 copies/μL in a blinded experiment with a high accuracy comparable with the qPCR. Its prototype consists of two disposable pen-like instruments for single-step sample preparation and contamination-free NATs, respectively. The simplified workflow of the FLASH enabled detection to be readily conducted by untrained users for at-home tests. All in all, the FLASH prototype demonstrates itself to be a promising home-use assay platform for effective mitigation of the pandemic.
快速居家检测是在疫情期间早期识别和管理感染者的一种很好的替代金标准定量聚合酶链反应(qPCR)的方法。然而,目前可用的居家快速抗原检测试剂盒和核酸检测(NAT)容易出现假结果。尽管一些基于CRISPR的NAT提高了准确性,但由于额外的开盖反应导致周转时间长(约1小时)和气溶胶污染,阻碍了其在自我检测中的应用。此外,由于缺乏样品纯化,居家NAT的准确性也受到样品基质干扰的影响。在此,我们报告了一种用于传染性病原体居家检测的快速、低成本、无气溶胶污染且灵敏的分子检测方法(FLASH),该方法结合了我们团队最近报道的等温且靶向特异性的NAT——oLAMP以及可视化侧向流动读数。该集成平台能够在20-30分钟内实现从样品到结果的SARS-CoV-2 RNA检测,在一项盲法实验中灵敏度达到0.5拷贝/μL,准确性与qPCR相当。其原型由两个一次性笔状仪器组成,分别用于单步样品制备和无污染物的NAT。FLASH简化的工作流程使未经培训的用户也能轻松在家中进行检测。总而言之,FLASH原型证明了自己是一个有前景的家用检测平台,可有效缓解疫情。