Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay

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

Published: Sept. 18, 2020

The recent outbreak of novel coronavirus (SARS-CoV-2) causing COVID-19 disease spreads rapidly in the world. Rapid and early detection SARS-CoV-2 facilitates intervention prevents spread. Here, we present an All-In-One Dual CRISPR-Cas12a (AIOD-CRISPR) assay for one-pot, ultrasensitive, visual detection. By targeting SARS-CoV-2's nucleoprotein gene, two CRISPR RNAs without protospacer adjacent motif (PAM) site limitation are introduced to develop AIOD-CRISPR detect nucleic acids with a sensitivity few copies. We validate by using clinical swab samples obtain consistent results RT-PCR assay. Furthermore, low-cost hand warmer (~$0.3) is used as incubator within 20 min, enabling instrument-free, at point care. Thus, our method has significant potential provide rapid, sensitive, one-pot point-of-care SARS-CoV-2.

Language: Английский

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Detection of unamplified target genes via CRISPR–Cas9 immobilized on a graphene field-effect transistor

Nature Biomedical Engineering, Journal Year: 2019, Volume and Issue: 3(6), P. 427 - 437

Published: March 25, 2019

Language: Английский

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Advancing Biosensors with Machine Learning

ACS Sensors, Journal Year: 2020, Volume and Issue: 5(11), P. 3346 - 3364

Published: Nov. 13, 2020

Chemometrics play a critical role in biosensors-based detection, analysis, and diagnosis. Nowadays, as branch of artificial intelligence (AI), machine learning (ML) have achieved impressive advances. However, novel advanced ML methods, especially deep learning, which is famous for image facial recognition, speech has remained relatively elusive to the biosensor community. Herein, how can be beneficial biosensors systematically discussed. The advantages drawbacks most popular algorithms are summarized on basis sensing data analysis. Specially, methods such convolutional neural network (CNN) recurrent (RNN) emphasized. Diverse ML-assisted electrochemical biosensors, wearable electronics, SERS other spectra-based fluorescence colorimetric comprehensively Furthermore, networks multibiosensor fusion introduced. This review will nicely bridge with greatly expand chemometrics

Language: Английский

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Loop-mediated isothermal amplification (LAMP) – review and classification of methods for sequence-specific detection

Analytical Methods, Journal Year: 2020, Volume and Issue: 12(6), P. 717 - 746

Published: Jan. 1, 2020

This comprehensive review provides a systematic classification and comparative evaluation of current sequence-specific detection methods for LAMP.

Language: Английский

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Transcriptome Profiling in Human Diseases: New Advances and Perspectives

International Journal of Molecular Sciences, Journal Year: 2017, Volume and Issue: 18(8), P. 1652 - 1652

Published: July 29, 2017

In the last decades, transcriptome profiling has been one of most utilized approaches to investigate human diseases at molecular level. Through expression studies, many biomarkers and therapeutic targets have found for several pathologies. This number is continuously increasing thanks total RNA sequencing. Indeed, this new technology completely revolutionized analysis allowing quantification gene levels allele-specific in a single experiment, as well identify novel genes, splice isoforms, fusion transcripts, world non-coding an unprecedented sequencing also employed important projects, like ENCODE (Encyclopedia regulatory elements) TCGA (The Cancer Genome Atlas), provide snapshot dozens cell lines thousands primary tumor specimens. Moreover, these studies paved way development data integration order facilitate management disease markers use clinics. scenario, ongoing clinical trials utilize through strategies instrument diagnosis numerous

Language: Английский

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All-in-One Dual CRISPR-Cas12a (AIOD-CRISPR) Assay: A Case for Rapid, Ultrasensitive and Visual Detection of Novel Coronavirus SARS-CoV-2 and HIV virus

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2020, Volume and Issue: unknown

Published: March 21, 2020

Abstract A recent outbreak of novel coronavirus (SARS-CoV-2), the causative agent COVID-19, has spread rapidly all over world. Human immunodeficiency virus (HIV) is another deadly and causes acquired syndrome (AIDS). Rapid early detection these viruses will facilitate intervention reduce disease transmission risk. Here, we present an ll- I n- O ne D ual CRISPR -Cas12a (termed “AIOD-CRISPR”) assay method for simple, rapid, ultrasensitive, one-pot, visual SARS-CoV-2 HIV virus. In our AIOD assay, a pair crRNAs was introduced to initiate dual CRISPR-Cas12a improve sensitivity. The AIOD-CRISPR system successfully utilized detect nucleic acids (DNA RNA) with sensitivity few copies. Also, it evaluated by detecting HIV-1 RNA extracted from human plasma samples, achieving comparable real-time RT-PCR method. Thus, great potential developing next-generation point-of-care molecular diagnostics.

Language: Английский

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CRISPR Systems for COVID-19 Diagnosis

ACS Sensors, Journal Year: 2021, Volume and Issue: 6(4), P. 1430 - 1445

Published: Jan. 27, 2021

The emergence of the new coronavirus 2019 (COVID-19) was first seen in December 2019, which has spread rapidly and become a global pandemic. number cases COVID-19 its associated mortality have raised serious concerns worldwide. Early diagnosis viral infection undoubtedly allows rapid intervention, disease management, substantial control disease. Currently, standard approach for globally is RT-qPCR test; however, limited access to kits reagents, need specialized lab equipment, highly skilled personnel led detection slowdown. Recently, development clustered regularly interspaced short palindromic repeats (CRISPR)-based diagnostic systems reshaped molecular diagnosis. benefits CRISPR system such as speed, precision, specificity, strength, efficiency, versatility inspired researchers develop CRISPR-based therapeutic methods. With outbreak, different groups begun design programs based on efficient system. advantages high speed (i.e., 30 min from raw sample reach result), sensitivity portability, no laboratory equipment. Here, we review contemporary studies

Language: Английский

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Present status of microfluidic PCR chip in nucleic acid detection and future perspective

TrAC Trends in Analytical Chemistry, Journal Year: 2022, Volume and Issue: 157, P. 116737 - 116737

Published: Aug. 11, 2022

Language: Английский

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Helicobacter pylori Infection, Its Laboratory Diagnosis, and Antimicrobial Resistance: a Perspective of Clinical Relevance

Clinical Microbiology Reviews, Journal Year: 2022, Volume and Issue: 35(3)

Published: April 11, 2022

Despite the recent decrease in overall prevalence of Helicobacter pylori infection, morbidity and mortality rates associated with gastric cancer remain high. The antimicrobial resistance developments treatment failure are fueling global burden H. pylori-associated complications. Accurate diagnosis remains opening move for eradication infections caused by microorganisms. Although several reports have been published on diagnostic approaches most lack data regarding from a clinical perspective. Therefore, we provide an intensive, comprehensive, updated description currently available methods that can help clinicians, infection professionals, researchers working epidemiology to broaden their understanding select appropriate methods. We also emphasize based settings (either or mass screening), patient factors age other predisposing factors), upper gastrointestinal bleeding partial gastrectomy) be considered evaluating efficacy. Furthermore, cope increasing trend resistance, better its emergence current detection inevitable.

Language: Английский

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Microfluidics-based strategies for molecular diagnostics of infectious diseases

Military Medical Research, Journal Year: 2022, Volume and Issue: 9(1)

Published: March 18, 2022

Abstract Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate point-of-care testing (POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology, potential substitute traditional methods in performing rapid, low-cost, accurate, on-site diagnoses. Molecular diagnostics widely used microfluidic devices as the most effective approaches pathogen detection. This review summarizes latest advances microfluidics-based molecular diseases from academic perspectives industrial outlooks. First, we introduce typical on-chip nucleic acid processes, including sample preprocessing, amplification, signal read-out. Then, four categories of platforms compared with respect to features, merits, demerits. We further discuss application digital assay absolute quantification. Both classic recent commercial summarized proof current market status. Finally, propose future directions diagnosis.

Language: Английский

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