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基于磁性纳米颗粒的简化病毒 RNA 提取方法,用于 SARS-CoV-2 的快速高通量检测。

A simplified viral RNA extraction method based on magnetic nanoparticles for fast and high-throughput detection of SARS-CoV-2.

机构信息

Materials Interfaces Center, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, PR China; University of Chinese Academy of Sciences, Beijing, 100049, PR China.

Materials Interfaces Center, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, PR China.

出版信息

Talanta. 2023 Jun 1;258:124479. doi: 10.1016/j.talanta.2023.124479. Epub 2023 Mar 23.

Abstract

The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based methods are being widely applied for clinical diagnosis, timely and accurate diagnosis towards COVID-19 causing virus, the SARS-CoV-2, is still limited due to labor-intensive and time-consuming operations. Herein, we report a new viral RNA extraction method based on poly-(amino ester) with carboxyl group (PC)-coated magnetic nanoparticles (pcMNPs) for the sensitive detection of SARS-CoV-2. This method combines the lysis and binding steps into one step, and refines multiple washing steps into one step, giving a turnaround time of less than 9 min. Furthermore, the extracted pcMNP-RNA complexes can be directly introduced into subsequent RT-PCR reactions without elution. This simplified viral RNA method could be well adapted in fast manual and automated high-throughput nucleic acids extraction protocols suitable for different scenarios. A high sensitivity down to 100 copies/mL and a linear correlation between 100 and 10 copies/mL of SARS-CoV-2 pseudovirus particles are achieved in both protocols. Benefitting from the simplicity and excellent performances, this new method can dramatically improve the efficiency and reduce operational requirements for the early clinical diagnosis and large-scale SARS-CoV-2 nucleic acid screening.

摘要

新型冠状病毒病 2019(COVID-19)的持续爆发引起了全球关注,因为其潜伏期长且具有很强的传染性。尽管基于 RT-PCR 的方法已广泛应用于临床诊断,但由于操作繁琐且耗时,对 COVID-19 致病病毒 SARS-CoV-2 的及时、准确诊断仍然有限。在此,我们报道了一种新的基于带羧基聚(氨基酯)(PC)修饰的磁性纳米粒子(pcMNPs)的病毒 RNA 提取方法,用于 SARS-CoV-2 的敏感检测。该方法将裂解和结合步骤合二为一,并将多个洗涤步骤合并为一步,总耗时不到 9 分钟。此外,提取的 pcMNP-RNA 复合物无需洗脱即可直接引入后续的 RT-PCR 反应。这种简化的病毒 RNA 提取方法可以很好地适应快速手动和自动化高通量核酸提取方案,适用于不同场景。两种方案均能达到 100 拷贝/mL 以下的高灵敏度和 100 至 10 拷贝/mL 之间的 SARS-CoV-2 假病毒颗粒的线性相关性。该方法简单、性能优异,可显著提高早期临床诊断和大规模 SARS-CoV-2 核酸筛查的效率,降低操作要求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cfa5/10035799/235aaf72367b/ga1_lrg.jpg

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