重组酶聚合酶扩增
清脆的
量油尺
微流控
聚合酶链反应
化学
计算生物学
核酸
多路复用
Cas9
桑格测序
PCR的应用
质粒
DNA
分子生物学
多重聚合酶链反应
纳米技术
DNA测序
生物
遗传学
基因
生物化学
材料科学
尿
作者
Hu Zhou,Zhichen Xu,Liang He,Zhijie Wang,Tao Zhang,Ting Hu,Fanwei Huang,Dongjuan Chen,Ying Li,Yunhuang Yang,Xiaoyuan Huang
标识
DOI:10.1021/acs.analchem.2c04713
摘要
Timely identification of human papillomavirus (HPV) infection is crucial for the prevention of cervical cancer. Current HPV detection methods mainly rely on polymerase chain reaction (PCR), which often requires bulky equipment and a long assay time. In this work, we report a heating-membrane-assisted multiplexed microfluidics platform that couples recombinase polymerase amplification (RPA) and CRISPR technology (termed M3-CRISPR) for fast and low-cost detection of multiple HPV subtypes. The heating membrane can provide convenient temperature control for the on-chip RPA and CRISPR assays. This stand-alone system allows simultaneous detection of HPV16 and HPV18 with high specificity and detection sensitivity (0.5 nM and 1 × 10–18 M for unamplified and amplified plasmids, respectively) in 30 min with a fluorescence-based readout. Furthermore, we introduced an optimized lateral flow dipstick (LFD) into the portable system to allow visualized detection of HPV DNA. The LFD-based readout also reached a detection sensitivity of 1 × 10–18 M for amplified plasmids and realized successful detection of HPV subtypes in the clinical samples. Finally, we established an automatic microfluidic system that enables the sample-in-answer-out detection of HPV subtypes. We believe that this fast, convenient, and affordable molecular diagnostic platform can serve as a useful tool in point-of-care testing of HPV or other pathogens.
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