CRISPR‐Enhanced Hydrogel Microparticles for Multiplexed Detection of Nucleic Acids DOI Creative Commons
Yoon Ho Roh, Chang Yeol Lee, Su-Jin Lee

et al.

Advanced Science, Journal Year: 2023, Volume and Issue: 10(10)

Published: Feb. 1, 2023

CRISPR/Cas systems offer a powerful sensing mechanism to transduce sequence-specific information into amplified analytical signals. However, performing multiplexed assays remains challenging and often requires complex approaches for assays. Here, hydrogel-based CRISPR/Cas12 system termed CLAMP (Cas-Loaded Annotated Micro-Particles) is described. The approach compartmentalizes the reaction in spatially-encoded hydrogel microparticles (HMPs). Each HMP identifiable by its face code becomes fluorescent when target DNA present. assay further streamlined capturing HMPs inside microfluidic device; captured particles are then automatically recognized machine-learning algorithm. fast, highly sensitive (attomolar detection limits with preamplification), capable of multiplexing single-pot assay. As proof-of-concept clinical application, applied detect nucleic acid targets human papillomavirus cervical brushing samples.

Language: Английский

Lift-CM: An integrated Lift-heater Centrifugal Microfluidic Platform for point-of-care pathogen nucleic acid detection using Isothermal amplification and CRISPR/Cas12a DOI

Taowei Shu,

Xueer Yin,

Qihua Xiong

et al.

Biosensors and Bioelectronics, Journal Year: 2025, Volume and Issue: 274, P. 117178 - 117178

Published: Jan. 18, 2025

Language: Английский

Citations

2
Enhanced chemiluminescence imaging sensor for ultrasensitive detection of nucleic acids based on HCR-CRISPR/Cas12a DOI

Xinxin Ke,

Yangjing Ou,

Yu Lin

et al.

Biosensors and Bioelectronics, Journal Year: 2022, Volume and Issue: 212, P. 114428 - 114428

Published: May 27, 2022

Language: Английский

Citations

59
Applications of surface‐enhanced Raman spectroscopy in environmental detection DOI Open Access
Lynn R. Terry,

Sage Sanders,

Rebecca H. Potoff

et al.

Analytical Science Advances, Journal Year: 2022, Volume and Issue: 3(3-4), P. 113 - 145

Published: March 23, 2022

As the human population grows, anthropogenic impacts from various agricultural and industrial processes produce unwanted contaminants in environment. The accurate, sensitive rapid detection of such is vital for health safety. Surface-enhanced Raman spectroscopy (SERS) a valuable analytical tool with wide applications environmental contaminant monitoring. aim this review to summarize recent advancements within SERS research as it applies detection, focus on published or accessible January 2021 through December including early-access publications. Our goal provide breadth information that can be used background knowledge field, well inform encourage further development techniques protecting quality Specifically, we highlight characteristics effective nanosubstrates, explore methods inorganic, organic, biological heavy metals, pharmaceuticals, plastic particles, synthetic dyes, pesticides, viruses, bacteria mycotoxins. We also discuss current limitations technologies propose several avenues future investigation. researchers fill identified gaps so implemented real-world environment more effectively efficiently, ultimately providing reliable timely data help make science-based strategies policies protect safety public health.

Language: Английский

Citations

58
Advanced Molecular and Immunological Diagnostic Methods to Detect SARS-CoV-2 Infection DOI Creative Commons
John Charles Rotondo, Fernanda Martini, Martina Maritati

et al.

Microorganisms, Journal Year: 2022, Volume and Issue: 10(6), P. 1193 - 1193

Published: June 10, 2022

COVID-19 emerged in late 2019 China and quickly spread across the globe, causing over 521 million cases of infection 6.26 deaths to date. After 2 years, numerous advances have been made. First all, preventive vaccine, which has implemented record time, is effective more than 95% cases. Additionally, diagnostic field, there are molecular antigenic kits that equipped with high sensitivity specificity. Real Time-PCR-based assays for detection viral RNA currently considered gold-standard method SARS-CoV-2 diagnosis can be used efficiently on pooled nasopharyngeal, or oropharyngeal samples widespread screening. Moreover, additional, advanced methods such as droplet-digital PCR (ddPCR), clustered regularly interspaced short palindromic repeats (CRISPR) next-generation sequencing (NGS), under development detect RNA. However, number subjects infected continuously increases globally, health care systems being placed increased stress. Thus, clinical laboratory plays an important role, helping select especially asymptomatic individuals who actively carrying live replicating virus, fast non-invasive technologies. Recent strategies, other methods, adopted either antigens, i.e., antigen-based immunoassays, human anti-SARS-CoV-2 antibodies, antibody-based nasal swabs, well blood saliva samples. role mucosal sIgAs, essential control viruses entering body through surfaces, remains elucidated, particular immune response counteracting infection, primarily at site(s) virus entry appears promising.

Language: Английский

Citations

53
CRISPR-Cas13a system: A novel tool for molecular diagnostics DOI Creative Commons

Lixin Zhao,

Minyue Qiu,

Xiaojia Li

et al.

Frontiers in Microbiology, Journal Year: 2022, Volume and Issue: 13

Published: Dec. 8, 2022

The clustered regularly interspaced short palindromic repeats (CRISPR) system is a natural adaptive immune of prokaryotes. CRISPR-Cas currently divided into two classes and six types: types I, III, IV in class 1 systems II, V, VI 2 systems. Among the type systems, CRISPR/Cas13a has been most widely characterized for its application molecular diagnostics, gene therapy, editing, RNA imaging. Moreover, because trans-cleavage activity Cas13a high specificity CRISPR RNA, enormous potential field diagnostics. Herein, we summarize applications detection pathogens, including viruses, bacteria, parasites, chlamydia, fungus; biomarkers, such as microRNAs, lncRNAs, circRNAs; some non-nucleic acid targets, proteins, ions, methyl groups. Meanwhile, highlight working principles novel Cas13a-based methods, Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) improved versions, nucleic amplification-free biosensors, biosensors target detection. Finally, focus on issues that need to be solved development prospects system.

Language: Английский

Citations

52
Recent advances in point of care testing for COVID-19 detection DOI Open Access
Renata Salgado Fernandes, Juliana de Oliveira Silva, Karina Braga Gomes

et al.

Biomedicine & Pharmacotherapy, Journal Year: 2022, Volume and Issue: 153, P. 113538 - 113538

Published: Aug. 12, 2022

The World Health Organizations declaration of the COVID-19 pandemic was a milestone for scientific community. high transmission rate and huge number deaths, along with lack knowledge about virus evolution disease, stimulated relentless search diagnostic tests, treatments, vaccines. main challenges were differential diagnosis development specific, rapid, sensitive tests that could reach all people. RT-PCR remains gold standard diagnosing COVID-19. However, new methods, such as other molecular techniques immunoassays emerged. Also, need accessible quick results boosted point care (POCT) are fast, automated, precision accuracy. This assay reduces dependence on laboratory conditions mass testing population, dispersing pressure regarding screening detection. review summarizes advances in field since started, emphasizing various detecting We reviewed existing well POCT under development, starting detection, but also exploring nucleic acid techniques, digital PCR, loop-mediated isothermal amplification-based (RT-LAMP), clustered regularly interspaced short palindromic repeats (CRISPR), next-generation sequencing (NGS), immunoassay nanoparticle-based biosensors, developed portable instruments rapid

Language: Английский

Citations

47
Microfluidics: the propellant of CRISPR-based nucleic acid detection DOI
Yanju Chen,

Siwenjie Qian,

Xiaoping Yu

et al.

Trends in biotechnology, Journal Year: 2022, Volume and Issue: 41(4), P. 557 - 574

Published: Aug. 18, 2022

Language: Английский

Citations

45
Sensitive Small Molecule Aptasensing based on Hybridization Chain Reaction and CRISPR/Cas12a Using a Portable 3D-Printed Visualizer DOI
Long Ma, Dan Liao,

Zhiying Zhao

et al.

ACS Sensors, Journal Year: 2023, Volume and Issue: 8(3), P. 1076 - 1084

Published: Jan. 18, 2023

Next-generation biosensing tools based on CRISPR/Cas have revolutionized the molecular detection. A number of CRISPR/Cas-based biosensors been reported for detection nucleic acid targets. The establishment efficient methods non-nucleic target would further broaden scope this technique, but up to now, concerning research is limited. In current study, we a versatile platform small-molecule called SMART-Cas12a (small-molecule aptamer regulated test using CRISPR/Cas12a). Simply, hybridization chain reaction cascade signal amplification was first trigged by functional (aptamer) through binding. Then, system integrated recognize amplified products followed activation trans-cleavage. As such, can be ingeniously converted signals and then fluorescent that readily visualized analyzed customized 3D-printed visualizer with help home-made App-enabled smartphone. Adenosine triphosphate selected as model target, under optimized conditions, achieved fine analytical performance linear range from 0.1 750 μM limit 1.0 nM. satisfactory selectivity recoveries obtained demonstrated method suitable complex sample environment. sample-to-answer time less than 100 min. Our work not only expanded reach CRISPR-Cas in also provided prototype generalized detecting wider analytes desirable adaptability, sensitivity, specificity, on-site capability.

Language: Английский

Citations

43
Sensitive and Amplification-Free Electrochemiluminescence Biosensor for HPV-16 Detection Based on CRISPR/Cas12a and DNA Tetrahedron Nanostructures DOI

Linying Yu,

Yao Peng,

Mengting Sheng

et al.

ACS Sensors, Journal Year: 2023, Volume and Issue: 8(7), P. 2852 - 2858

Published: July 4, 2023

Rapid and accurate detection of biomarkers was very important for early screening treatment diseases. Herein, a sensitive amplification-free electrochemiluminescence (ECL) biosensor based on CRISPR/Cas12a DNA tetrahedron nanostructures (TDNs) constructed. Briefly, 3D TDN self-assembled the Au nanoparticle-deposited glassy carbon electrode surface to construct biosensing interface. The presence target would activate trans-cleavage activity Cas12a-crRNA duplex cleave single-stranded signal probe vertex TDN, causing Ru(bpy)32+ fall from weakened ECL signal. Thus, system transduced change concentration into an enabling HPV-16. specific recognition HPV-16 made have good selectivity, while TDN-modified sensing interface could reduce cleaving steric resistance improve performance CRISPR/Cas12a. In addition, pretreated complete sample within 100 min with limit 8.86 fM, indicating that developed possesses potential application prospect fast nucleic acid detection.

Language: Английский

Citations

42
CRISPR-based biosensors for pathogenic biosafety DOI
Hao Yang, Rodrigo Ledesma‐Amaro, Hong Gao

et al.

Biosensors and Bioelectronics, Journal Year: 2023, Volume and Issue: 228, P. 115189 - 115189

Published: March 2, 2023

Language: Английский

Citations

41