Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing.
Capillary-based reverse transcriptase loop-mediated isothermal amplification for cost-effective and rapid point-of-care COVID-19 testing.
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DOI：
10.1016/j.aca.2021.339002
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发表时间：
2021-11-15
影响因子：
6.2
通讯作者：
Sones CL
中科院分区：
文献类型：
--
作者：
John AJUK;He PJW;Katis IN;Galanis PP;Iles AH;Eason RW;Sones CL
随着SARS-CoV-2大流行的继续蔓延，快速、简便的诊断能力的必要性从未如此重要。考虑到这一目标，我们开发了一种具有成本效益和节省时间的检测方法/策略，该方法/策略在狭窄的、市售的和廉价的玻璃毛细管内实施灵敏的逆转录酶环介导扩增（RT-LAMP）检测，用于检测SARS-CoV-2病毒RNA。该方法与广泛使用的基于实验室的分子检测方案和当前可用的基础设施兼容。它采用一种简单的快速提取方案，裂解病毒，释放足够的遗传物质进行扩增。然后使用SARS-CoV-2 RT-LAMP试剂盒在恒定温度下扩增该提取的病毒RNA，所得扩增产物产生可目视判读的颜色变化。该测试方案与RT-LAMP测定结合，具有样品的每次反应100个病毒拷贝的灵敏度，并在略超过30分钟内提供结果。由于该分析是在大多数测试实验室常用的水浴中进行的，因此无需专门的仪器和相关技能。此外，我们的测试途径需要显著减少每次测试的试剂量，同时提供与本研究中使用的RT-LAMP试剂盒相当的灵敏度和特异性。传统技术需要25 μl试剂，而我们的测试仅使用不到一半的量（10 μl）。因此，凭借其简约的方法，这种基于毛细管的检测可能是传统检测的一种有前途的替代方法，因为它可以在资源有限的环境中使用现成的设备进行，并且具有增加整体检测能力的潜力，同时也减轻了大规模检测供应链的负担。
As the SARS-CoV-2 pandemic continues to spread, the necessity for rapid, easy diagnostic capabilities could never have been more crucial. With this aim in mind, we have developed a cost-effective and time-saving testing methodology/strategy that implements a sensitive reverse transcriptase loop-mediated amplification (RT-LAMP) assay within narrow, commercially available and cheap, glass capillaries for detection of the SARS-CoV-2 viral RNA. The methodology is compatible with widely used laboratory-based molecular testing protocols and currently available infrastructure. It employs a simple rapid extraction protocol that lyses the virus, releasing sufficient genetic material for amplification. This extracted viral RNA is then amplified using a SARS-CoV-2 RT-LAMP kit, at a constant temperature and the resulting amplified product produces a colour change which can be visually interpreted. This testing protocol, in conjunction with the RT-LAMP assay, has a sensitivity of ∼100 viral copies per reaction of a sample and provides results in a little over 30 min. As the assay is carried out in a water bath, commonly available within most testing laboratories, it eliminates the need for specialised instruments and associated skills. In addition, our testing pathway requires a significantly reduced quantity of reagents per test while providing comparable sensitivity and specificity to the RT-LAMP kit used in this study. While the conventional technique requires 25 μl of reagent, our test only utilises less than half the quantity (10 μl). Thus, with its minimalistic approach, this capillary-based assay could be a promising alternative to the conventional testing, owing to the fact that it can be performed in resource-limited settings, using readily available apparatus, and has the potential of increasing the overall testing capacity, while also reducing the burden on supply chains for mass testing.
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影响因子：
7.7
作者：
Yang Q;Meyerson NR;Clark SK;Paige CL;Fattor WT;Gilchrist AR;Barbachano-Guerrero A;Healy BG;Worden-Sapper ER;Wu SS;Muhlrad D;Decker CJ;Saldi TK;Lasda E;Gonzales P;Fink MR;Tat KL;Hager CR;Davis JC;Ozeroff CD;Brisson GR;McQueen MB;Leinwand LA;Parker R;Sawyer SL
通讯作者：
Sawyer SL
影响因子：
5.7
作者：
Huang, Wei E.;Lim, Boon;Cui, Zhanfeng
通讯作者：
Cui, Zhanfeng
通过逆转录环介导的等温扩增测定快速直观地检测 2019 新型冠状病毒 (SARS-CoV-2)
影响因子：
14.2
作者：
Yan, C.;Cui, J.;Yuan, J.
通讯作者：
Yuan, J.
DOI：
10.3390/v12090972
发表时间：
2020-09-01
期刊：
Viruses
影响因子：
--
作者：
Rohaim MA;Clayton E;Sahin I;Vilela J;Khalifa ME;Al-Natour MQ;Bayoumi M;Poirier AC;Branavan M;Tharmakulasingam M;Chaudhry NS;Sodi R;Brown A;Burkhart P;Hacking W;Botham J;Boyce J;Wilkinson H;Williams C;Whittingham-Dowd J;Shaw E;Hodges M;Butler L;Bates MD;La Ragione R;Balachandran W;Fernando A;Munir M
通讯作者：
Munir M
影响因子：
7.4
作者：
Bwambok, David K.;Christodouleas, Dionysios C.;Whitesides, George M.
通讯作者：
Whitesides, George M.