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一种快速且可及的用于分离RNA、DNA和蛋白质以促进新冠病毒检测的方法

A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2.

作者信息

Ponce-Rojas Jose Carlos, Costello Michael S, Proctor Duncan A, Kosik Kenneth S, Wilson Maxwell Z, Arias Carolina, Acosta-Alvear Diego

机构信息

Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, Santa Barbara, California, USA.

Neuroscience Research Institute, Santa Barbara, California, USA.

出版信息

J Clin Microbiol. 2021 Mar 19;59(4). doi: 10.1128/JCM.02403-20.

Abstract

Management of the coronavirus disease 2019 (COVID-19) pandemic requires widespread testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among them RNA extraction kits. The need for commercial RNA extraction kits places a bottleneck on tests that detect SARS-CoV-2 genetic material, including PCR-based reference tests. Here, we propose an alternative method we call PEARL (recipitation-nhanced nalyte etrieva) that addresses this limitation. PEARL uses a lysis solution that disrupts cell membranes and viral envelopes while simultaneously providing conditions suitable for alcohol-based precipitation of RNA, DNA, and proteins. PEARL is a fast, low-cost, and simple method that uses common laboratory reagents and offers performance comparable to that of commercial RNA extraction kits. PEARL offers an alternative method to isolate host and pathogen nucleic acids and proteins to streamline the detection of DNA and RNA viruses, including SARS-CoV-2.

摘要

2019冠状病毒病(COVID-19)大流行的管理需要对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)进行广泛检测。广泛开展SARS-CoV-2检测的一个主要限制因素是全球基本物资短缺,其中包括RNA提取试剂盒。对商用RNA提取试剂盒的需求给检测SARS-CoV-2遗传物质的检测带来了瓶颈,包括基于PCR的参考检测。在此,我们提出了一种名为PEARL(沉淀增强分析物提取)的替代方法,以解决这一限制。PEARL使用一种裂解液,该裂解液可破坏细胞膜和病毒包膜,同时提供适合基于酒精沉淀RNA、DNA和蛋白质的条件。PEARL是一种快速、低成本且简单的方法,它使用常见的实验室试剂,其性能与商用RNA提取试剂盒相当。PEARL提供了一种分离宿主和病原体核酸及蛋白质的替代方法,以简化包括SARS-CoV-2在内的DNA和RNA病毒的检测。

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