PD-LAMP smartphone detection of SARS-CoV-2 on chip.
PD-LAMP smartphone detection of SARS-CoV-2 on chip.
复制标题
PD-LAMP智能手机芯片检测SARS-CoV-2。
DOI：
10.1016/j.aca.2022.339702
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发表时间：
2022-04-22
影响因子：
6.2
通讯作者：
Kinzer-Ursem TL
中科院分区：
文献类型：
--
作者：
Colbert AJ;Lee DH;Clayton KN;Wereley ST;Linnes JC;Kinzer-Ursem TL
2019 年，由 SARS-CoV-2 引起的 COVID-19 大流行证明了对快速、可靠和便携式诊断的迫切需求。 2020 年 1 月宣布 COVID-19 大流行，疫情持续激增。显然，早期识别感染者，特别是无症状携带者，对于预防疾病传播发挥着巨大作用。目前诊断 SARS-CoV-2 的黄金标准是基于病毒 RNA 检测的定量逆转录聚合酶链反应 (qRT-PCR) 测试。虽然 RT-PCR 可靠且灵敏，但它需要昂贵的集中设备并且耗时（∼2 小时或更长）；限制了其在资源匮乏地区的适用性。 FDA 为多种 COVID-19 诊断颁发了紧急使用授权 (EUA)，重点是护理点 (PoC) 测试。许多 RT-PCR 和血清学检测被批准在护理点使用。 Abbott 的 ID NOW 和 Cue Health 的 COVID-19 检测特别令人感兴趣，它们使用等温扩增方法在 20 分钟内快速检测。我们希望通过新型快速便携式等温核酸检测设备来扩展当前 PoC 测试平台的范围。我们将逆转录环介导的等温扩增 (RT-LAMP) 与粒子成像技术、粒子扩散测定法 (PD) 结合起来，在集成加热功能的便携式芯片上仅用 35 分钟即可成功检测 SARS-CoV-2。智能手机设备用于对 RT-LAMP 后含有荧光珠的样品进行成像，并将扩散率降低与阳性样品相关联。我们使用便携式加热装置从微流控芯片中的 RT-LAMP 反应中检测到每 μL 少至 30 个病毒颗粒。此外，我们可以对稀释的未处理唾液样本进行 RT-LAMP，无需提取 RNA。此外，我们还冻干了微流控芯片中针对 N 基因和 ORF1ab 基因的 SARS-CoV-2 特异性 RT-LAMP 反应，从而无需冷藏。我们的检测方法符合世界卫生组织概述的特定目标产品概况：它针对 SARS-CoV-2，不需要冷藏，与数字连接兼容，并且唾液中的检测限低于每毫升 35 × 104 个病毒颗粒。 PD-LAMP 快速、简单，对于在护理点进行筛查和使用具有吸引力。
In 2019 the COVID-19 pandemic, caused by SARS-CoV-2, demonstrated the urgent need for rapid, reliable, and portable diagnostics. The COVID-19 pandemic was declared in January 2020 and surges of the outbreak continue to reoccur. It is clear that early identification of infected individuals, especially asymptomatic carriers, plays a huge role in preventing the spread of the disease. The current gold standard diagnostic for SARS-CoV-2 is quantitative reverse transcription polymerase chain reaction (qRT-PCR) test based on the detection of the viral RNA. While RT-PCR is reliable and sensitive, it requires expensive centralized equipment and is time consuming (∼2 h or more); limiting its applicability in low resource areas. The FDA issued Emergency Use Authorizations (EUAs) for several COVID-19 diagnostics with an emphasis on point-of care (PoC) testing. Numerous RT-PCR and serological tests were approved for use at the point of care. Abbott's ID NOW, and Cue Health's COVID-19 test are of particular interest, which use isothermal amplification methods for rapid detection in under 20 min. We look to expand on the range of current PoC testing platforms with a new rapid and portable isothermal nucleic acid detection device. We pair reverse transcription loop mediated isothermal amplification (RT-LAMP) with a particle imaging technique, particle diffusometry (PD), to successfully detect SARS-CoV-2 in only 35 min on a portable chip with integrated heating. A smartphone device is used to image the samples containing fluorescent beads post-RT-LAMP and correlates decreased diffusivity to positive samples. We detect as little as 30 virus particles per μL from a RT-LAMP reaction in a microfluidic chip using a portable heating unit. Further, we can perform RT-LAMP from a diluted unprocessed saliva sample without RNA extraction. Additionally, we lyophilize SARS-CoV-2-specific RT-LAMP reactions that target both the N gene and the ORF1ab gene in the microfluidic chip, eliminating the need for cold storage. Our assay meets specific target product profiles outlined by the World Health Organization: it is specific to SARS-CoV-2, does not require cold storage, is compatible with digital connectivity, and has a detection limit of less than 35 × 104 viral particles per mL in saliva. PD-LAMP is rapid, simple, and attractive for screening and use at the point of care.
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影响因子：
1.2
作者：
Chen, Hua-Wei;Weissenberger, Giulia;Ching, Wei-Mei
通讯作者：
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作者：
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影响因子：
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作者：
Hou, Tieying;Zeng, Weiqi;Xu, Teng
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影响因子：
12.6
作者：
Moehling TJ;Lee DH;Henderson ME;McDonald MK;Tsang PH;Kaakeh S;Kim ES;Wereley ST;Kinzer-Ursem TL;Clayton KN;Linnes JC
通讯作者：
Linnes JC