Recent advances in RNA sample preparation techniques for the detection of SARS-CoV-2 in saliva and gargle

Liu, Yanming; Kumblathan, Teresa; Tao, Jeffrey; Xu, Jingyang; Feng, Wei; Xiao, Huyan; Hu, Jianyu; Huang, Camille; Wu, Yiping; Zhang, Hongquan; Li, Xing‐Fang; Le, X. Chris

doi:10.1016/j.trac.2023.117107

Cited by 7 publications

(2 citation statements)

References 113 publications

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“…We also applied the CRISPR/Cas13a-responsive hydrogel capillary sensor to the detection of SARS-CoV-2 RNA in saliva samples because previous work has demonstrated the promising potential of saliva for POC diagnostic applications. − We analyzed 10 heat-treated saliva samples, comprising five samples (saliva 1−5) from confirmed SARS-CoV-2-infected patients and five specimens (saliva 6−10) from healthy individuals. As displayed in Figure D, all the SARS-CoV-2 positive saliva sample solutions generated significant travel distances in the capillaries compared to the negative saliva samples and the blank control.…”

Section: Results and Discussionmentioning

confidence: 99%

CRISPR/Cas13a-Responsive and RNA-Bridged DNA Hydrogel Capillary Sensor for Point-of-Care Detection of RNA

Wang,

Pian

et al. 2024

Anal. Chem.

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Disease diagnostics and surveillance increasingly highlight the importance of portable, cost-effective, and sensitive point-of-care (POC) detection of nucleic acids. Here, we report a CRISPR/Cas13a-responsive and RNA-bridged DNA hydrogel capillary sensor for the direct and visual detection of specific RNA with high sensitivity. The capillary sensor was simply prepared by loading RNA-cross-linking DNA hydrogel film (∼0.2 mm ± 0.02 mm) at the end of a capillary. When CRISPR/Cas13a specifically recognizes the target RNA, the RNA bridge in the hydrogel film is cleaved by the trans-cleavage activity of CRISPR/Cas13a, increasing the permeability of the hydrogel film. Different concentrations of target RNA activate different amounts of Cas13a, cleaving different amounts of the RNA bridge in the hydrogel and causing corresponding changes in the permeability of the hydrogel. Therefore, samples containing different amounts of the target RNA travel to different distances in the capillary. Visual reading of the distance provides quantitative detection of the RNA target without the need for any nucleic acid amplification or auxiliary equipment. The technique was successfully used for the determination of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in clinical nasopharyngeal (NP) swab and saliva samples. Easily quantifiable distance using a ruler eliminates the need for any optical or electrochemical detection equipment, making this assay potentially useful for POC and on-site applications.

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Section: Results and Discussionmentioning

confidence: 99%

CRISPR/Cas13a-Responsive and RNA-Bridged DNA Hydrogel Capillary Sensor for Point-of-Care Detection of RNA

Wang,

Pian

et al. 2024

Anal. Chem.

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“…One of them is to design effective sample preparation and detection methods that can extract, concentrate, and purify the target analytes (e.g., nucleic acids or proteins) from the bodily fluids, detect their presence, and/or measure their quantity in a simple, rapid, and accurate way. 11 Sample preparation and detection methods are crucial steps in any diagnostic test, as they can affect the accuracy, sensitivity, and specificity of the test results. To address these challenges, many research efforts have been carried out as reported in the literature.…”

Section: Introductionmentioning

confidence: 99%

Sample preparation and detection methods in point-of-care devices towards future at-home testing

Adedokun,

Alipanah,

Fan

2024

Lab Chip

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Integrated Antigenic and Nucleic Acid Detection in Single Virions and Extracellular Vesicles with Viral Content

Nguyen,

Rima,

Nguyen

et al. 2024

Adv Healthcare Materials

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Virion‐mediated outbreaks are imminent and despite rapid responses, continue to cause adverse symptoms and death. Therefore, tunable, sensitive, high‐throughput assays are needed to help diagnose future virion‐mediated outbreaks. Herein, we developed a tunable in situ assay to selectively enrich virions and extracellular vesicles (EVs) and simultaneously detect antigens and nucleic acids at a single‐particle resolution. The Biochip Antigen and RNA Assay (BARA) enhanced sensitivities compared to quantitative reverse‐transcription polymerase chain reaction (qRT‐PCR), enabling the detection of virions in asymptomatic patients, genetic mutations in single virions, and enabling the continued long‐term expression of viral RNA in the EV‐enriched subpopulation in the plasma of patients with post‐acute sequelae of COVID‐19. BARA revealed highly accurate diagnoses of COVID‐19 by simultaneously detecting the spike glycoprotein and nucleocapsid‐encoding RNA in saliva and nasopharyngeal swab samples. Altogether, the single‐particle detection of antigens and viral RNA provides a tunable framework for the diagnosis, monitoring, and mutation screening of current and future outbreaks.This article is protected by copyright. All rights reserved

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Recent advances in RNA sample preparation techniques for the detection of SARS-CoV-2 in saliva and gargle

Cited by 7 publications

References 113 publications

CRISPR/Cas13a-Responsive and RNA-Bridged DNA Hydrogel Capillary Sensor for Point-of-Care Detection of RNA

CRISPR/Cas13a-Responsive and RNA-Bridged DNA Hydrogel Capillary Sensor for Point-of-Care Detection of RNA

Sample preparation and detection methods in point-of-care devices towards future at-home testing

Integrated Antigenic and Nucleic Acid Detection in Single Virions and Extracellular Vesicles with Viral Content

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