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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. REVEALR: A Multicomponent XNAzyme-Based Nucleic Acid Detection System for SARS-CoV-2

Author

John C. Chaput and Kefan Yang

Subjects
Male, Paper, Analyte, Loop-mediated isothermal amplification, Computational biology, 010402 general chemistry, Sensitivity and Specificity, 01 natural sciences, Biochemistry, Catalysis, chemistry.chemical_compound, Colloid and Surface Chemistry, Limit of Detection, Nasopharynx, Chlorocebus aethiops, Animals, Humans, CRISPR, Vero Cells, Detection limit, SARS-CoV-2, COVID-19, RNA, DNA, Catalytic, General Chemistry, Nucleic acid amplification technique, 0104 chemical sciences, Oligodeoxyribonucleotides, chemistry, COVID-19 Nucleic Acid Testing, Nucleic acid, RNA, Viral, Female, Nucleic Acid Amplification Techniques, DNA
Abstract

Isothermal amplification strategies capable of rapid, inexpensive, and accurate nucleic acid detection provide new options for large-scale pathogen detection, disease diagnosis, and genotyping. Here we report a highly sensitive multicomponent XNA-based nucleic acid detection platform that combines analyte preamplification with X10-23-mediated catalysis to detect the viral pathogen responsible for COVID-19. The platform, termed RNA-Encoded Viral Nucleic Acid Analyte Reporter (REVEALR), functions with a detection limit of ≤20 aM (∼10 copies/μL) using conventional fluorescence and paper-based lateral flow readout modalities. With a total assay time of 1 h, REVEALR provides a convenient nucleic acid alternative to equivalent CRISPR-based approaches, which have become popular methods for SARS-CoV-2 detection. The assay shows no cross-reactivity for other in vitro transcribed respiratory viral RNAs and functions with perfect accuracy against COVID-19 patient-derived clinical samples.

Published
2021

3. Engineering Paper-Based Sensors for Zika Virus

Author

Oscar A. Negrete, Robert J. Meagher, and Koen K. A. Van Rompay

Subjects
0301 basic medicine, Paper, Cas9, Zika Virus Infection, Diagnostic test, Paper based, Zika Virus, Biology, Microfluidic Analytical Techniques, Molecular diagnostics, biology.organism_classification, Virology, Data science, Southeast asia, Zika virus, 03 medical and health sciences, 030104 developmental biology, CRISPR, Humans, RNA, Viral, Molecular Medicine, CRISPR-Cas Systems, Molecular Biology
Abstract

The emergence of Zika virus (ZIKV) infections in Latin America and Southeast Asia has created an urgent need for new, simple, yet sensitive, diagnostic tests. We highlight recent work using paper-based sensors coupled with CRISPR/Cas9 to detect ZIKV RNA as a new approach to achieve rapid development and deployment of field-ready diagnostics for emerging infectious diseases.

Published
2016
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