用于在20分钟内快速检测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)提取RNA的手持式即时检测系统。

Handheld Point-of-Care System for Rapid Detection of SARS-CoV-2 Extracted RNA in under 20 min.

作者信息

Rodriguez-Manzano Jesus, Malpartida-Cardenas Kenny, Moser Nicolas, Pennisi Ivana, Cavuto Matthew, Miglietta Luca, Moniri Ahmad, Penn Rebecca, Satta Giovanni, Randell Paul, Davies Frances, Bolt Frances, Barclay Wendy, Holmes Alison, Georgiou Pantelis

机构信息

Department of Infectious Disease, Faculty of Medicine, Imperial College London, London SW7 2AZ, United Kingdom.

Department of Electrical and Electronic Engineering, Faculty of Engineering, Imperial College London, London SW7 2AZ, United Kingdom.

出版信息

ACS Cent Sci. 2021 Feb 24;7(2):307-317. doi: 10.1021/acscentsci.0c01288. Epub 2021 Jan 13.

Abstract

The COVID-19 pandemic is a global health emergency characterized by the high rate of transmission and ongoing increase of cases globally. Rapid point-of-care (PoC) diagnostics to detect the causative virus, SARS-CoV-2, are urgently needed to identify and isolate patients, contain its spread and guide clinical management. In this work, we report the development of a rapid PoC diagnostic test (<20 min) based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) and semiconductor technology for the detection of SARS-CoV-2 from extracted RNA samples. The developed LAMP assay was tested on a real-time benchtop instrument (RT-qLAMP) showing a lower limit of detection of 10 RNA copies per reaction. It was validated against extracted RNA from 183 clinical samples including 127 positive samples (screened by the CDC RT-qPCR assay). Results showed 91% sensitivity and 100% specificity when compared to RT-qPCR and average positive detection times of 15.45 ± 4.43 min. For validating the incorporation of the RT-LAMP assay onto our PoC platform (RT-eLAMP), a subset of samples was tested ( = 52), showing average detection times of 12.68 ± 2.56 min for positive samples ( = 34), demonstrating a comparable performance to a benchtop commercial instrument. Paired with a smartphone for results visualization and geolocalization, this portable diagnostic platform with secure cloud connectivity will enable real-time case identification and epidemiological surveillance.

摘要

新冠疫情是一场全球卫生突发事件,其特点是传播速度快且全球病例持续增加。迫切需要快速即时检测(PoC)诊断方法来检测致病病毒严重急性呼吸综合征冠状病毒2(SARS-CoV-2),以识别和隔离患者、遏制其传播并指导临床管理。在本研究中,我们报告了一种基于逆转录环介导等温扩增(RT-LAMP)和半导体技术的快速PoC诊断测试(<20分钟)的开发,用于从提取的RNA样本中检测SARS-CoV-2。所开发的LAMP检测方法在实时台式仪器(RT-qLAMP)上进行了测试,显示每个反应的检测下限为10个RNA拷贝。它针对从183份临床样本(包括127份阳性样本,由美国疾病控制与预防中心RT-qPCR检测筛选)中提取的RNA进行了验证。与RT-qPCR相比,结果显示敏感性为91%,特异性为100%,平均阳性检测时间为15.45±4.43分钟。为了验证将RT-LAMP检测方法整合到我们的PoC平台(RT-eLAMP)上的效果,对一部分样本(n = 52)进行了测试,结果显示阳性样本(n = 34)的平均检测时间为12.68±2.56分钟,证明其性能与台式商用仪器相当。该便携式诊断平台与智能手机配对用于结果可视化和地理定位,并具有安全的云连接功能,将能够实现实时病例识别和流行病学监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4087/7908026/6fd8706d4359/oc0c01288_0001.jpg

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