Liu Juxiang, Chen Ping, Hu Xulong, Huang Liping, Geng Zhi, Xu Hao, Hu Wenjun, Wang Lin, Wu Ping, Liu Gang L
National Engineering Research Center for Nanomedicine, College of Life Science and Technology, Huazhong University of Science and Technology, 1037 Luo Yu Road, Wuhan 430074, China.
Institute of Geophysics and Geomatics, China University of Geosciences, Wuhan 430074, China.
Chem Eng J. 2023 Jan 1;451:138822. doi: 10.1016/j.cej.2022.138822. Epub 2022 Aug 28.
The novel mutations attributed by the high mutagenicity of the SARS-CoV-2 makes its prevention and treatment challenging. Developing an ultra-fast, point-of-care-test (POCT) protocol is critical for responding to large-scale spread of SARS-CoV-2 in public places and in resource-poor remote areas. Here, we developed a nanoplasmonic enhanced isothermal amplification (NanoPEIA) strategy that combines a nanoplasmonic sensor with isothermal amplification. The novel strategy provides an ideal easy-to operate detection platform for obtaining accurate, ultra-fast and high-throughput (96 samples can be tested together) data. For clinical samples with viral detection at Ct value <25, the entire process (including sample preparation, virus lysis, detection, and data analysis) can be completed within six minutes. The method is also appropriate for detection of SARS-CoV-2 -coronavirus mutants. The NanoPEIA method was validated using clinical samples from 21 patients with SARS-CoV-2 infection and 31 healthy individuals. The detection result on the 52 clinical samples for SARS-CoV-2 showed that the NanoPEIA platform had a 100% sensitivity for N and orf1ab genes, which was higher than those obtained using RT-qPCR (88.9% and 90.0%, respectively). The specificities of 31 clinical negative samples were 92.3% and 91.7% for the N gene and the orf1ab gene, respectively. The limits of detection (LoD) of the clinical samples were 28.3 copies/mL and 23.3 copies/mL for the N gene and the orf1ab gene, respectively. The efficient NanoPEIA detection strategy facilitates real-time detection and visualization within ultrashort durations and can be applied for POCT diagnosis in resource-poor and highly populated areas.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的高致突变性所产生的新突变使其预防和治疗具有挑战性。开发一种超快速的即时检测（POCT）方案对于应对SARS-CoV-2在公共场所和资源匮乏的偏远地区的大规模传播至关重要。在此，我们开发了一种纳米等离子体增强等温扩增（NanoPEIA）策略，该策略将纳米等离子体传感器与等温扩增相结合。这种新策略提供了一个理想的易于操作的检测平台，可用于获取准确、超快速和高通量（可同时检测96个样本）的数据。对于病毒检测Ct值<25的临床样本，整个过程（包括样本制备、病毒裂解、检测和数据分析）可在6分钟内完成。该方法也适用于检测SARS-CoV-2冠状病毒突变体。使用来自21例SARS-CoV-2感染患者和31名健康个体的临床样本对NanoPEIA方法进行了验证。对52份SARS-CoV-2临床样本的检测结果表明，NanoPEIA平台对N基因和orf1ab基因的灵敏度均为100%，高于逆转录定量聚合酶链反应（RT-qPCR）的检测结果（分别为88.9%和90.0%）。31份临床阴性样本对N基因和orf1ab基因的特异性分别为92.3%和91.7%。临床样本对N基因和orf1ab基因的检测限分别为28.3拷贝/毫升和23.3拷贝/毫升。高效的NanoPEIA检测策略有助于在超短时间内进行实时检测和可视化，可应用于资源匮乏和人口密集地区的POCT诊断。
