A universal all-in-one RPA-Cas12a strategy with de novo autodesigner and its application in on-site ultrasensitive detection of DNA and RNA viruses
A universal all-in-one RPA-Cas12a strategy with de novo autodesigner and its application in on-site ultrasensitive detection of DNA and RNA viruses
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DOI：
10.1016/j.bios.2023.115609
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发表时间：
2023-08-21
影响因子：
12.6
通讯作者：
Huang, Lizhen
中科院分区：
文献类型：
--
作者：
Lin, Cailing;Chen, Feng;Huang, Lizhen
革命性的一体化RPA-CRISPR检测由于其高灵敏度和易用性，正迅速成为最受欢迎的即时检测（POCT）工具。尽管针对特定目标的一锅法是可用的，但针对新目标开发更有效的方法仍然是一个重大挑战。在这项研究中，我们提出了一种快速和通用的方法来建立基于合理平衡扩增和Cas 12 a切割的一体化RPA-Cas 12 a方法CORDSv 2，该方法实现了对包括SARS-CoV-2，ASFV，HPV 16和HPV 18在内的几种靶标的超灵敏检测。CORDSv 2的检测限（LOD）为0.6 cp/mu L，对SARS-CoV-2的灵敏度为100%，与qPCR相当。与便携式检测仪（SARS-CORDS）联用，检测限为6cp/μ L，对SARS-CoV-2和Ct的灵敏度分别为100%和97
Revolutionary all-in-one RPA-CRISPR assays are rapidly becoming the most sought-after tools for point-of-care testing (POCT) due to their high sensitivity and ease of use. Despite the availability of one-pot methods for specific targets, the development of more efficient methods for new targets remains a significant challenge. In this study, we present a rapid and universal approach to establishing an all-in-one RPA-Cas12a method CORDSv2 based on rational balancing amplification and Cas12a cleavage, which achieves ultrasensitive detection of several targets, including SARS-CoV-2, ASFV, HPV16, and HPV18. CORDSv2 demonstrates a limit of detection (LOD) of 0.6 cp/mu L and 100% sensitivity for SARS-CoV-2, comparable to qPCR. Combining with our portable device(hippo-CORDS), it has a visual detection LOD of 6 cp/mu L and a sensitivity up to 100% for SARS-CoV-2 and 97% for Ct