Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong Special Administrative Region of China.
Department of Medicine & Therapeutics, The Chinese University of Hong Kong, Shatin, Hong Kong Special Administrative Region of China.
Biosens Bioelectron. 2025 Jan 1;267:116866. doi: 10.1016/j.bios.2024.116866. Epub 2024 Oct 22.
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.
CoRPLA（CRISPR 调控的一站式重组酶聚合酶环介导扩增）是一种基于扩增子耗尽、皮温操作、适用于家庭检测的 iNAAT。它使用专门设计的环引物来增强等温扩增，触发 Cas12 进行原位扩增子耗尽和信号放大。该方法解决了扩增子衍生气溶胶污染和复杂检测方案等问题，实现了亚皮摩尔灵敏度的定量检测（0.5 cps/μL）。CoRPLA 采用 DNA 水凝胶可穿戴胶带进行实时比色读数，可直观地区分病原体载量。它已通过 SARS-CoV-2、RSV、流感 A 和 HPV 的临床样本进行了验证，成功地识别了阳性病例的多层次病毒载量，结果与 qPCR 一致。CoRPLA 在消除气溶胶污染假阳性的同时保持高灵敏度，将分子检测从台式仪器推向家庭，用于日常病毒感染监测。
