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1. CRISPR/Cas14 and G-Quadruplex DNAzyme-Driven Biosensor for Paper-Based Colorimetric Detection of African Swine Fever Virus.

Author

Zhao, Xue, He, Yawen, Shao, Shengjie, Ci, Qiaoqiao, Chen, Lin, Lu, Xiaonan, Liu, Qian, and Chen, Juhong

Subjects
African swine fever virus, CRISPR, Cas14, G-quadruplex, microfluidic paper-based analytical device, pathogen detection, African Swine Fever Virus, Colorimetry, Biosensing Techniques, DNA, Catalytic, G-Quadruplexes, Paper, Animals, CRISPR-Cas Systems, Swine, DNA, Viral, Limit of Detection
Abstract

The highly contagious nature and 100% fatality rate contribute to the ongoing and expanding impact of the African swine fever virus (ASFV), causing significant economic losses worldwide. Herein, we developed a cascaded colorimetric detection using the combination of a CRISPR/Cas14a system, G-quadruplex DNAzyme, and microfluidic paper-based analytical device. This CRISPR/Cas14a-G4 biosensor could detect ASFV as low as 5 copies/μL and differentiate the wild-type and mutated ASFV DNA with 2-nt difference. Moreover, this approach was employed to detect ASFV in porcine plasma. A broad linear detection range was observed, and the limit of detection in spiked porcine plasma was calculated to be as low as 42-85 copies/μL. Our results indicate that the developed paper platform exhibits the advantages of high sensitivity, excellent specificity, and low cost, making it promising for clinical applications in the field of DNA disease detection and suitable for popularization in low-resourced areas.

Published
2024

2. Timing readout in paper device for quantitative point-of-use hemin/G-quadruplex DNAzyme-based bioassays

Author

Jinfang Nie, Shangwang Le, Yun Zhang, Wenyuan Zhu, Dong Gao, Jiani Yang, Songbai Zhang, Jianping Li, and Jinlong Fan

Subjects
Paper, Analyte, Specific detection, Point-of-Care Systems, Microfluidics, Biomedical Engineering, Biophysics, Deoxyribozyme, Nanotechnology, Biosensing Techniques, G-quadruplex, chemistry.chemical_compound, Limit of Detection, Electrochemistry, Humans, Reproducibility of Results, DNA, Catalytic, General Medicine, Serum samples, G-Quadruplexes, chemistry, Potassium, Hemin, Biological Assay, Timer, DNA Probes, Biotechnology
Abstract

This work describes a quantitative point-of-use hemin/G-quadruplex DNAzyme-based assay that integrates a simple timing detection motif with low-cost, portable microfluidic paper-based analytical devices (μPADs). The timing readout is based on the selective DNAzyme-mediated wettability change of paper from hydrophilicity to hydrophobicity. Its utility is well demonstrated with sensitive, specific detection of K(+) ion as a model analyte in artificial samples as well as real human serum samples. This new method only requires a ubiquitous cheap timer (or a cell phone with a timing function) to provide quantitative results. It could offer new opportunities for the development of more peroxidase-mimicking DNAzyme-based bioassays that are simple, affordable, portable, and operable by minimally-trained users for broad point-of-use applications especially in resource-poor settings.

Published
2015
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