Ultrasensitive CRISPR-based diagnostic for field-applicable detection of Plasmodium species in symptomatic and asymptomatic malaria DOI Creative Commons
Rose Lee,

Helena de Puig,

Peter Q. Nguyen

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2020, Volume and Issue: 117(41), P. 25722 - 25731

Published: Sept. 21, 2020

Significance Detection of submicroscopic malaria in asymptomatic individuals is needed for eradication and remains a diagnostic gap resource-limited settings. Nonfalciparum clinical diagnostics are second gap, as these infections have low parasite density commonly undetected. We describe an integrated, 60-min, ultrasensitive specific CRISPR-based the four major pathogenic Plasmodium species that can fill gaps. Using SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform, we designed assays with limits detection below recommended by World Health Organization. These simplified sample preparation method: rapid extraction protocol, which eliminates complicated nucleic acid steps. Our work further translates platform into field-deployable diagnostic.

Language: Английский

Diagnosing COVID-19: The Disease and Tools for Detection DOI Creative Commons

Buddhisha Udugama,

Pranav Kadhiresan,

H Kozłowski

et al.

ACS Nano, Journal Year: 2020, Volume and Issue: 14(4), P. 3822 - 3835

Published: March 30, 2020

COVID-19 has spread globally since its discovery in Hubei province, China December 2019. A combination of computed tomography imaging, whole genome sequencing, and electron microscopy were initially used to screen identify SARS-CoV-2, the viral etiology COVID-19. The aim this review article is inform audience diagnostic surveillance technologies for SARS-CoV-2 their performance characteristics. We describe point-of-care diagnostics that are on horizon encourage academics advance beyond conception. Developing plug-and-play manage outbreak would be useful preventing future epidemics.

Language: Английский

Citations

1749
CRISPR-based diagnostics DOI Open Access
Michael M. Kaminski, Omar O. Abudayyeh, Jonathan S. Gootenberg

et al.

Nature Biomedical Engineering, Journal Year: 2021, Volume and Issue: 5(7), P. 643 - 656

Published: July 16, 2021

Language: Английский

Citations

845
Diagnostics for SARS-CoV-2 infections DOI Creative Commons
Bhavesh D. Kevadiya, Jatin Machhi, Jonathan Herskovitz

et al.

Nature Materials, Journal Year: 2021, Volume and Issue: 20(5), P. 593 - 605

Published: Feb. 15, 2021

Language: Английский

Citations

736
Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA DOI Creative Commons
Maturada Patchsung,

Krittapas Jantarug,

Archiraya Pattama

et al.

Nature Biomedical Engineering, Journal Year: 2020, Volume and Issue: 4(12), P. 1140 - 1149

Published: Aug. 26, 2020

Language: Английский

Citations

564
SARS-CoV-2 RapidPlex: A Graphene-Based Multiplexed Telemedicine Platform for Rapid and Low-Cost COVID-19 Diagnosis and Monitoring DOI Creative Commons
Rebeca M. Torrente‐Rodríguez, Heather Lukas, Jiaobing Tu

et al.

Matter, Journal Year: 2020, Volume and Issue: 3(6), P. 1981 - 1998

Published: Oct. 5, 2020

Language: Английский

Citations

451
Ultrasensitive and Highly Specific Lateral Flow Assays for Point-of-Care Diagnosis DOI Creative Commons
Yilin Liu, Li Zhan, Zhenpeng Qin

et al.

ACS Nano, Journal Year: 2021, Volume and Issue: 15(3), P. 3593 - 3611

Published: Feb. 19, 2021

Lateral flow assays (LFAs) are paper-based point-of-care (POC) diagnostic tools that widely used because of their low cost, ease use, and rapid format. Unfortunately, traditional commercial LFAs have significantly poorer sensitivities (μM) specificities than standard laboratory tests (enzyme-linked immunosorbent assay, ELISA: pM-fM; polymerase chain reaction, PCR: aM), thus limiting impact in disease control. In this Perspective, we review the evolving efforts to increase sensitivity specificity LFAs. Recent work improve through assay improvement includes optimization kinetics signal amplification by either reader systems or additional reagents. Together, these produced with ELISA-level (pM-fM). addition, sample preamplification can be applied both nucleic acids (direct amplification) other analytes (indirect prior LFA testing, which lead PCR-level (aM) sensitivity. However, strategies also detection time complexity, inhibits large-scale POC use Perspectives achieve future (<30 min), ultrasensitive (PCR-level), "sample-to-answer" diagnostics provided. case specificity, recent research focused on high-affinity molecules reduce nonspecific binding. Furthermore, novel highly specific molecules, such as CRISPR/Cas systems, integrated into diagnosis produce not only but diagnostics. summary, continuing improvements, may soon offer performance at is competitive techniques while retaining a

Language: Английский

Citations

424
Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities DOI Open Access
Jeanne E. van Dongen, Johanna Theodora Wilhelmina Berendsen, Renske D.M. Steenbergen

et al.

Biosensors and Bioelectronics, Journal Year: 2020, Volume and Issue: 166, P. 112445 - 112445

Published: July 26, 2020

Language: Английский

Citations

321
Nucleic Acid Detection Using CRISPR/Cas Biosensing Technologies DOI
Rashid Aman, Ahmed Mahas, Magdy M. Mahfouz

et al.

ACS Synthetic Biology, Journal Year: 2020, Volume and Issue: 9(6), P. 1226 - 1233

Published: March 11, 2020

For infectious diseases, rapid and accurate identification of the pathogen is critical for effective management treatment, but diagnosis remains challenging, particularly in resource-limited areas. Methods that accurately detect nucleic acids can provide robust, accurate, rapid, ultrasensitive technologies point-of-care pathogens, thus yield information invaluable disease treatment. Several technologies, mostly PCR-based, have been employed detection; however, these require expensive reagents equipment, skilled personnel. CRISPR/Cas systems used genome editing, based on their ability to recognize cleave specific DNA RNA sequences. Moreover, following recognition target sequence, certain including orthologues Cas13, Cas12a, Cas14 exhibit collateral nonspecific catalytic activities be acid detection, example by degradation a labeled produce fluorescent signal. are amenable multiplexing, thereby enabling single diagnostic test identify multiple targets down attomolar (10–18 mol/L) concentrations molecules. Developing devices couple with lateral flow may allow inexpensive, highly sensitive, in-field deployable diagnostics. These sensors myriad applications, from human health agriculture. In this review, we discuss recent advances field CRISPR-based biosensing highlight insights potential use applications.

Language: Английский

Citations

316
Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection DOI Creative Commons
Long Thành Nguyễn, Brianna M. Smith, Piyush Jain

et al.

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: Sept. 30, 2020

Abstract The CRISPR-Cas12a RNA-guided complexes have tremendous potential for nucleic acid detection but are limited to the picomolar limit without an amplification step. Here, we develop a platform with engineered crRNAs and optimized conditions that enabled us detect various clinically relevant targets higher sensitivity, achieving of in femtomolar range any target pre-amplification By extending 3′- or 5′-ends crRNA different lengths ssDNA, ssRNA, phosphorothioate discover self-catalytic behavior augmented rate LbCas12a-mediated collateral cleavage activity as high 3.5-fold compared wild-type significant improvement specificity recognition. Particularly, 7-mer DNA extension is determined be universal spacer-independent enhancing sensitivity detection. We perform detailed characterization our ENHANCE system modifications, types, reporters, divalent cations. With isothermal SARS-CoV-2 RNA using RT-LAMP, modified incorporated paper-based lateral flow assay can up 23-fold within 40–60 min.

Language: Английский

Citations

316
A CRISPR/Cas12a-empowered surface plasmon resonance platform for rapid and specific diagnosis of the Omicron variant of SARS-CoV-2 DOI Creative Commons
Zhi Chen, Jingfeng Li, Tianzhong Li

et al.

National Science Review, Journal Year: 2022, Volume and Issue: 9(8)

Published: June 3, 2022

The outbreak of the COVID-19 pandemic was partially due to challenge identifying asymptomatic and presymptomatic carriers virus, thus highlights a strong motivation for diagnostics with high sensitivity that can be rapidly deployed. On other hand, several concerning SARS-CoV-2 variants, including Omicron, are required identified as soon samples 'positive'. Unfortunately, traditional PCR test does not allow their specific identification. Herein, first time, we have developed MOPCS (Methodologies Photonic CRISPR Sensing), which combines an optical sensing technology-surface plasmon resonance (SPR) 'gene scissors' clustered regularly interspaced short palindromic repeat (CRISPR) technique achieve both specificity when it comes measurement viral variants. is low-cost, CRISPR/Cas12a-system-empowered SPR gene-detecting platform analyze RNA, without need amplification, within 38 min from sample input results output, limit detection 15 fM. achieves highly sensitive analysis SARS-CoV-2, mutations appear in variants B.1.617.2 (Delta), B.1.1.529 (Omicron) BA.1 (a subtype Omicron). This also used some recently collected patient local China, by Centers Disease Control Prevention. innovative CRISPR-empowered will further contribute fast, accurate target nucleic acid sequences single-base mutations.

Language: Английский

Citations

301