Evaluation of recombinase-based isothermal amplification assays for point-of-need detection of SARS-CoV-2 in resource-limited settings.
Evaluation of recombinase-based isothermal amplification assays for point-of-need detection of SARS-CoV-2 in resource-limited settings.
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DOI：
10.1016/j.ijid.2021.11.007
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发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Mondal D
中科院分区：
文献类型：
--
作者：
Ghosh P;Chowdhury R;Hossain ME;Hossain F;Miah M;Rashid MU;Baker J;Rahman MZ;Rahman M;Ma X;Duthie MS;Wahed AAE;Mondal D
诊断的民主化是遏制 2019 年冠状病毒病 (COVID-19) 在全球范围内传播的关键挑战之一。现有基于 PCR 的方法操作复杂，包括将样本转移到配备昂贵设备的先进中心实验室，限制了它们在资源有限的环境中的使用。然而，随着等温技术的出现，在分散的设施中检测严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 是可能的。在本研究中，评估了两种基于重组酶的等温技术，即逆转录重组酶聚合酶扩增（RT-RPA）和逆转录重组酶辅助扩增（RT-RAA），用于检测临床样本中的 SARS-CoV-2。总共评估了 76 例实时逆转录 PCR（实时 RT-PCR）确诊的 COVID-19 病例和 100 例阴性对照，以确定等温方法的诊断性能。这项研究表明，两种方法的诊断准确性同样有希望，当 RT-RPA 和 RT-RAA 的靶基因分别为 RdRP 和 ORF1ab 时，敏感性为 76.32%（95% 置信区间 65.18–85.32%）； RT-RPA 中 N 和 RdRP 的组合提高了检测的准确性，灵敏度为 85.53%（95% 置信区间 75.58–92.55%）。此外，每种方法的特异性都很高，范围为 94.00% 至 98.00%（95% 置信区间为 87.40–9.76%）。考虑到诊断准确性，RT-RPA 和 RT-RAA 似乎都是适合按需部署的检测方法，用于检测病原体、了解其流行病学、病例管理和遏制传播。
The democratization of diagnostics is one of the key challenges towards containing the transmission of coronavirus disease 2019 (COVID-19) around the globe. The operational complexities of existing PCR-based methods, including sample transfer to advanced central laboratories with expensive equipment, limit their use in resource-limited settings. However, with the advent of isothermal technologies, the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is possible at decentralized facilities. In this study, two recombinase-based isothermal techniques, reverse transcription recombinase polymerase amplification (RT-RPA) and reverse transcription recombinase-aided amplification (RT-RAA), were evaluated for the detection of SARS-CoV-2 in clinical samples. A total of 76 real-time reverse transcription PCR (real-time RT-PCR) confirmed COVID-19 cases and 100 negative controls were evaluated to determine the diagnostic performance of the isothermal methods. This investigation revealed equally promising diagnostic accuracy of the two methods, with a sensitivity of 76.32% (95% confidence interval 65.18–85.32%) when the target genes were RdRP and ORF1ab for RT-RPA and RT-RAA, respectively; the combination of N and RdRP in RT-RPA augmented the accuracy of the assay at a sensitivity of 85.53% (95% confidence interval 75.58–92.55%). Furthermore, high specificity was observed for each of the methods, ranging from 94.00% to 98.00% (95% confidence interval 87.40–9.76%). Considering the diagnostic accuracies, both RT-RPA and RT-RAA appear to be suitable assays for point-of-need deployment for the detection of the pathogen, understanding its epidemiology, case management, and curbing transmission.
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影响因子：
3.2
作者：
Mondal D;Ghosh P;Khan MA;Hossain F;Böhlken-Fascher S;Matlashewski G;Kroeger A;Olliaro P;Abd El Wahed A
通讯作者：
Abd El Wahed A
用于病毒检测的增强的等温扩增测定法。
影响因子：
16.6
作者：
Qian J;Boswell SA;Chidley C;Lu ZX;Pettit ME;Gaudio BL;Fajnzylber JM;Ingram RT;Ward RH;Li JZ;Springer M
通讯作者：
Springer M
DOI：
10.5354/0719-0527.2020.60648
发表时间：
2020-01-01
影响因子：
0.1
作者：
Nassehi, Armin
通讯作者：
Nassehi, Armin
影响因子：
9.3
作者：
Behrmann, Ole;Bachmann, Iris;Hufert, Frank T.
通讯作者：
Hufert, Frank T.
影响因子：
8.8
作者：
Abd El Wahed, Ahmed;Weidmann, Manfred;Hufert, Frank T.
通讯作者：
Hufert, Frank T.