Review of analytical performance of COVID-19 detection methods

Analytical and Bioanalytical Chemistry, Journal Year: 2020, Volume and Issue: 413(1), P. 35 - 48

Published: Sept. 18, 2020

Abstract In the recent SARS-CoV-2 pandemic, public health experts have emphasized testing, tracking infected people, and tracing their contacts as an effective strategy to reduce spread of virus. Several diagnostic methods are reported for detecting coronavirus in clinical, research, laboratories. Some tests detect infection directly by viral RNA other indirectly host antibodies. A test during pandemic should help make appropriate clinical decision a short period time. Recently varying throughput, batching capacity, requirement infrastructure setting, analytical performance, turnaround times ranging from few minutes several hours. These factors be considered while selecting reliable rapid method prompt interventions. This paper reviews published journals reports released regulatory agencies. We compared efficiency including limit detection, sensitivity, specificity, throughput. addition, we also looked into ease use, affordability, availability accessories. Finally, discuss limitations provide our perspectives on priorities future development.

Language: Английский

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CRISPR-Cas based virus detection: Recent advances and perspectives

Biosensors and Bioelectronics, Journal Year: 2021, Volume and Issue: 193, P. 113541 - 113541

Published: Aug. 8, 2021

Language: Английский

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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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An engineered CRISPR-Cas12a variant and DNA-RNA hybrid guides enable robust and rapid COVID-19 testing

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: March 19, 2021

Extensive testing is essential to break the transmission of SARS-CoV-2, which causes ongoing COVID-19 pandemic. Here, we present a CRISPR-based diagnostic assay that robust viral genome mutations and temperature, produces results fast, can be applied directly on nasopharyngeal (NP) specimens without RNA purification, incorporates human internal control within same reaction. Specifically, show use an engineered AsCas12a enzyme enables detection wildtype mutated SARS-CoV-2 allows us perform step with loop-mediated isothermal amplification (LAMP) at 60-65 °C. We also find hybrid DNA-RNA guides increases rate reaction, enabling our test completed 30 minutes. Utilizing clinical samples from 72 patients infection 57 healthy individuals, demonstrate exhibits specificity positive predictive value 100% sensitivity 50 1000 copies per reaction (or 2 40 microliter) for purified unpurified NP respectively.

Language: Английский

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Droplet Cas12a Assay Enables DNA Quantification from Unamplified Samples at the Single-Molecule Level

Nano Letters, Journal Year: 2021, Volume and Issue: 21(11), P. 4643 - 4653

Published: May 26, 2021

DNA quantification is important for biomedical research, but the routinely used techniques rely on nucleic acid amplification which have inherent issues like cross-contamination risk and bias. Here, we report a CRISPR-Cas12a-based molecular diagnostic technique amplification-free absolute of at single-molecule level. To achieve this, first screened out optimal reaction parameters high-efficient Cas12a assay, yielding over 50-fold improvement in sensitivity compared with reported assays. We further leveraged microdroplet-enabled confinement effect to perform an ultralocalized droplet obtaining excellent specificity sensitivity. Moreover, demonstrated its versatility capability by direct counting diverse virus's DNAs (African swine fever virus, Epstein–Barr Hepatitis B virus) from clinical serum samples wide range viral titers. Given flexible programmability crRNA, envision this as versatile quantitative platform diagnosis.

Language: Английский

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Amplification-free CRISPR/Cas detection technology: challenges, strategies, and perspectives

Chemical Society Reviews, Journal Year: 2022, Volume and Issue: 52(1), P. 361 - 382

Published: Dec. 19, 2022

This review summarizes the recent advances and main strategies to improve sensitivity of amplification-free CRISPR/Cas-based detection techniques.

Language: Английский

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Photocontrolled crRNA activation enables robust CRISPR-Cas12a diagnostics

Proceedings of the National Academy of Sciences, Journal Year: 2022, Volume and Issue: 119(26)

Published: June 21, 2022

CRISPR diagnostics based on nucleic acid amplification faces barriers to its commercial use, such as contamination risks and insufficient sensitivity. Here, we propose a robust solution involving optochemical control of RNA (crRNA) activation in detection. Based this strategy, recombinase polymerase (RPA) CRISPR-Cas12a detection systems can be integrated into completely closed test tube. crRNA designed temporarily inactivated so that RPA is not affected by Cas12a cleavage. After the reaction completed, system activated under rapid light irradiation. This photocontrolled, fully diagnostic avoids exhibits more than two orders magnitude improvement sensitivity compared with conventional one-pot assay. photocontrolled method was applied clinical severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA, achieving specificity comparable those PCR. Furthermore, compact automatic device constructed.

Language: Английский

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CRISPR Enzyme Kinetics for Molecular Diagnostics

Analytical Chemistry, Journal Year: 2021, Volume and Issue: 93(20), P. 7456 - 7464

Published: May 12, 2021

CRISPR-diagnostic assays have gained significant interest in the last few years. This has grown rapidly during current COVID-19 pandemic, where CRISPR-diagnostics been frontline contenders for rapid testing solutions. surge research prompts following question: what exactly are achievable limits of detection and associated assay times enabled by kinetics enzymes such as Cas12 Cas13? To explore this question, we here present a model based on Michaelis–Menten enzyme theory applied to CRISPR enzymes. We use develop analytical solutions reaction back-of-the-envelope criteria validate check consistency reported kinetic parameters. our analyses all studies known us, which report Michaelis–Menten-type data CRISPR-associated These include subtypes Cas13 orthologs. found but one study clearly violate at least two three rules therefore that basic physical limits. performed an experimental LbCas12a with both ssDNA dsDNA activators these its predicted scaling. The validated is used time scales degree completion practically relevant target concentrations applicable assays. results broad implications emerging, amplification-free CRISPR-detection methods.

Language: Английский

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CRISPR/Cas12a-based technology: A powerful tool for biosensing in food safety

Trends in Food Science & Technology, Journal Year: 2022, Volume and Issue: 122, P. 211 - 222

Published: March 1, 2022

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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