Benchtop to at-home test: Amplicon-depleted CRISPR-regulated loop mediated amplification at skin-temperature for viral load monitoring.

IF 12.7 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

ACS Central Science Pub Date : 2025-01-01 Epub Date: 2024-10-22 DOI:10.1016/j.bios.2024.116866

Yumeng Cao, Haosi Lin, Xiao Lu, Xiaolong Wu, Yuxuan Zhu, Zibin Zhao, Yanan Li, Samantha Borje, Grace C Y Lui, Shui Shan Lee, Hnin Yin Yin Nyein, I-Ming Hsing

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引用次数: 0

Abstract

CoRPLA (CRISPR-regulated One-pot Recombinase Polymerase Loop-mediated Amplification) is an amplicon-depleted skin-temperature operated iNAAT designed for at-home testing. It uses specially designed loop primers to enhance isothermal amplification, triggering Cas12 for in-situ amplicon depletion and signal amplification. This method addresses issues like amplicon-derived aerosol contamination and complex assay formats, enabling quantitative detection with sub-attomolar sensitivity (0.5 cps/μL). CoRPLA employs a DNA hydrogel wearable tape for real-time, colorimetric readout, allowing visual differentiation of pathogen loads. It was validated with clinical samples for SARS-CoV-2, RSV, influenza A, and HPV, successfully identifying multi-level viral loads of the positive cases with results consistent with qPCR. Offering high sensitivity while eliminating false positives from aerosol contamination, CoRPLA bridges the molecular assay from benchtop to home for daily viral infections monitoring.

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从台式测试到家用测试：用于病毒载量监测的皮肤温度下的扩增片段删除 CRISPR 调节环介导扩增。

CoRPLA（CRISPR-regulated One-pot Recombinase Polymerase Loop-mediated Amplification，CRISPR调控单锅重组酶聚合酶环介导扩增）是一种扩增片段耗尽皮肤温度操作的iNAAT，设计用于家庭检测。它使用专门设计的环路引物来增强等温扩增，触发 Cas12 进行原位扩增片段耗竭和信号扩增。该方法解决了扩增子产生的气溶胶污染和复杂的检测格式等问题，实现了亚阿托摩尔灵敏度（0.5 cps/μL）的定量检测。CoRPLA 采用 DNA 水凝胶可穿戴胶带进行实时比色读数，可直观区分病原体负荷。它通过临床样本对 SARS-CoV-2、RSV、甲型流感和 HPV 进行了验证，成功鉴定了阳性病例的多级病毒载量，结果与 qPCR 一致。CoRPLA 具有高灵敏度，同时消除了气溶胶污染造成的假阳性，为日常病毒感染监测架起了从台式到家用的分子检测桥梁。

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来源期刊

ACS Central Science Chemical Engineering-General Chemical Engineering

CiteScore

25.50

自引率

0.50%

发文量

194

审稿时长

10 weeks

期刊介绍： ACS Central Science publishes significant primary reports on research in chemistry and allied fields where chemical approaches are pivotal. As the first fully open-access journal by the American Chemical Society, it covers compelling and important contributions to the broad chemistry and scientific community. "Central science," a term popularized nearly 40 years ago, emphasizes chemistry's central role in connecting physical and life sciences, and fundamental sciences with applied disciplines like medicine and engineering. The journal focuses on exceptional quality articles, addressing advances in fundamental chemistry and interdisciplinary research.