Advanced Point‐of‐Care Testing Technologies for Human Acute Respiratory Virus Detection

Advanced Materials, Journal Year: 2021, Volume and Issue: 34(1)

Published: Oct. 8, 2021

Abstract The ever‐growing global threats to human life caused by the acute respiratory virus (RV) infections have cost billions of lives, created a significant economic burden, and shaped society for centuries. timely response emerging RVs could save lives reduce medical care burden. development RV detection technologies is essential potentially preventing pandemic epidemics. However, commonly used lack sensitivity, specificity, speed, thus often failing provide rapid turnaround times. To address this problem, new are devised performance inadequacies traditional methods. These offer improvements in convenience, flexibility, portability point‐of‐care test (POCT). Herein, recent developments POCT comprehensively reviewed eight typical viruses. This review discusses challenges opportunities various recognition strategies these according their principles, including nucleic acid amplification, optical POCT, electrochemistry, lateral flow assays, microfluidics, enzyme‐linked immunosorbent microarrays. importance limits detection, throughput, portability, specificity when testing clinical samples resource‐limited settings emphasized. Finally, evaluation commercial kits both diagnosis clinical‐oriented practices included.

Language: Английский

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The CRISPR–Cas toolbox for analytical and diagnostic assay development

Chemical Society Reviews, Journal Year: 2021, Volume and Issue: 50(21), P. 11844 - 11869

Published: Jan. 1, 2021

A comprehensive review that offers mechanistic insight into the CRISPR–Cas toolbox for analytical and diagnostic assay development.

Language: Английский

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Glycerol Additive Boosts 100-fold Sensitivity Enhancement for One-Pot RPA-CRISPR/Cas12a Assay

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(23), P. 8277 - 8284

Published: May 30, 2022

CRISPR/Cas12, a highly efficient and specific nucleic acid recognition system, has been broadly employed to detect amplified DNA products. However, most reported methods adopt two-step detection mode that needs liquid transfer step, thus complicating the procedure posing risk of aerosol contamination. A one-pot method can obviate these problems, but it suffers from poor efficiency due loss amplification templates elicited by CRISPR/Cas12 cleavage. In this study, we discovered glycerol additive dramatically promoted recombinase polymerase (RPA)-CRISPR/Cas12a method. Compared with glycerol-free version, its sensitivity was nearly 100-fold higher close canonical Further investigation displayed enhanced attributed phase separation RPA CRISPR/Cas12a system during initial reaction caused viscosity. This triumphantly harnessed for African swine fever virus (ASFV) SARS-CoV-2, achieving naked-eye readout through smartphone-equipped device. The currently developed glycerol-enhanced RPA-CRISPR/Cas12a be an advantageous point-of-care platform on account simplicity, high sensitivity, universality.

Language: Английский

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Signal amplification and output of CRISPR/Cas-based biosensing systems: A review

Analytica Chimica Acta, Journal Year: 2021, Volume and Issue: 1185, P. 338882 - 338882

Published: July 26, 2021

Language: Английский

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CRISPR‐Cas Biochemistry and CRISPR‐Based Molecular Diagnostics

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 62(17)

Published: Jan. 30, 2023

Abstract Polymerase chain reaction (PCR)‐based nucleic acid testing has played a critical role in disease diagnostics, pathogen surveillance, and many more. However, this method requires long turnaround time, expensive equipment, trained personnel, limiting its widespread availability diagnostic capacity. On the other hand, clustered regularly interspaced short palindromic repeats (CRISPR) technology recently demonstrated capability for detection with high sensitivity specificity. CRISPR‐mediated biosensing holds great promise revolutionizing procedures developing point‐of‐care diagnostics. This review focuses on recent developments both fundamental CRISPR biochemistry CRISPR‐based techniques. Four ongoing research hotspots molecular diagnostics‐target preamplification‐free detection, microRNA (miRNA) testing, non‐nucleic‐acid SARS‐CoV‐2 detection‐are also covered.

Language: Английский

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Programmable RNA detection with CRISPR-Cas12a

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Sept. 5, 2023

Abstract Cas12a, a CRISPR-associated protein complex, has an inherent ability to cleave DNA substrates and is utilized in diagnostic tools identify molecules. We demonstrate that multiple orthologs of Cas12a activate trans-cleavage the presence split activators. Specifically, PAM-distal region crRNA recognizes RNA targets provided PAM-proximal seed target. Our method, Split Activator for Highly Accessible Analysis (SAHARA), detects picomolar concentrations without sample amplification, reverse-transcription, or strand-displacement by simply supplying short sequence complementary region. Beyond detection, SAHARA outperforms wild-type CRISPR-Cas12a specificity towards point-mutations can detect pooled crRNA/Cas12a arrays via distinct DNAs. In conclusion, simple, yet powerful nucleic acid detection platform based on be applied multiplexed fashion potentially expanded other CRISPR-Cas enzymes.

Language: Английский

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Enzyme Kinetics and Detector Sensitivity Determine Limits of Detection of Amplification-Free CRISPR-Cas12 and CRISPR-Cas13 Diagnostics

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(27), P. 9826 - 9834

Published: June 27, 2022

Interest in CRISPR-Cas12 and CRISPR-Cas13 detection continues to increase as these schemes enable the specific recognition of nucleic acids. The fundamental sensitivity limits (and their applicability amplification-free assays) are governed by kinetic rates. However, rates remain poorly understood, reporting has been inconsistent. We quantify parameters for several enzymes (LbCas12a, AsCas12a, AapCas12b, LwaCas13a, LbuCas13a) corresponding (LoD). Collectively, we present quantification enzyme kinetics 14 guide RNAs (gRNAs) acid targets a total 50 sets rate 25 LoDs. validate self-consistency our measurements comparing trends limiting behaviors with Michaelis–Menten trans-cleavage reaction model. For assay conditions, activated Cas12 Cas13 exhibit catalytic efficiencies between order 105 106 M–1 s–1. assays that use fluorescent reporter molecules (ssDNA ssRNA) target detection, at current conditions result an LoD picomolar range. results suggest successful requires cleavage (by CRISPR enzyme) least 0.1% molecules. This fraction reporters cleaved is required differentiate signal from background, hypothesize this largely independent method (e.g., endpoint vs velocity) detector sensitivity. Our demonstrate nature which background limit LoDs thus highlight areas improvement emerging field diagnostics.

Language: Английский

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Argonaute-integrated isothermal amplification for rapid, portable, multiplex detection of SARS-CoV-2 and influenza viruses

Biosensors and Bioelectronics, Journal Year: 2022, Volume and Issue: 207, P. 114169 - 114169

Published: March 18, 2022

Language: Английский

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Recent development of surface-enhanced Raman scattering for biosensing

Journal of Nanobiotechnology, Journal Year: 2023, Volume and Issue: 21(1)

Published: May 6, 2023

Abstract Surface-Enhanced Raman Scattering (SERS) technology, as a powerful tool to identify molecular species by collecting spectral signals at the single-molecule level, has achieved substantial progresses in fields of environmental science, medical diagnosis, food safety, and biological analysis. As deepening research is delved into SERS sensing, more high-performance or multifunctional substrate materials emerge, which are expected push sensing application fields. Especially field analysis, intrinsic extrinsic schemes have been widely used explored due their fast, sensitive reliable advantages. Herein, recent developments substrates applications biomolecular detection (SARS-CoV-2 virus, tumor etc.), imaging pesticide summarized. The concepts (including its basic theory mechanism) important strategies (extending from nanomaterials with tunable shapes nanostructures surface bio-functionalization modifying affinity groups specific biomolecules) for improving biosensing performance comprehensively discussed. For data analysis identification, machine learning methods software acquisition sources diagnosing discussed detail. In conclusion, challenges perspectives future presented.

Language: Английский

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CLIPON: A CRISPR‐Enabled Strategy that Turns Commercial Pregnancy Test Strips into General Point‐of‐Need Test Devices

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(12)

Published: Jan. 22, 2022

Abstract Desirable biosensing assays need to be sensitive, specific, cost‐effective, instrument‐free, and versatile. Herein we report a new strategy termed CLIPON ( C RISPR L arge DNA assembly I nduced P regnancy strips for signal‐ ON detection) that can deliver these traits. integrates commercial pregnancy test strip (PTS) with four biological elements: the human chorionic gonadotropin (hCG), CRISPR‐Cas12a, crRNA c auliflower‐like l arge‐sized D NA assemblies (CLD). uses Cas12a/crRNA complex both recognize target of interest release CLD‐bound hCG so presence translate into colorimetric signal on PTS. We demonstrate versatility through sensitive specific detection HPV genomic DNA, SARS‐CoV‐2 RNA adenosine. also engineer cell phone app hand‐held microchip achieve quantification. represents an attractive option point‐of‐care diagnostics.

Language: Английский

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