CRISPR-Cas12a Biosensor Array for Ultrasensitive Detection of Unamplified DNA with Single-Nucleotide Polymorphic Discrimination DOI

Zhengyan Weng,

Zheng You, Huijie Li

et al.

ACS Sensors, Journal Year: 2023, Volume and Issue: 8(4), P. 1489 - 1499

Published: April 7, 2023

Quantitative polymerase chain reaction as a powerful tool for DNA detection has been pivotal to vast range of applications, including disease screening, food safety assessment, environmental monitoring, and many others. However, the essential target amplification step in combination with fluorescence readout poses significant challenge rapid streamlined analysis. The discovery engineering clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated (Cas) technology have recently paved way novel approach nucleic acid detection, but majority current CRISPR-mediated platforms are limited by insufficient sensitivity still require preamplification. Herein, we report CRISPR-Cas12a-mediated graphene field-effect transistor (gFET) array, named CRISPR Cas12a-gFET, amplification-free, ultrasensitive, reliable both single-stranded (ssDNA) double-stranded (dsDNA) targets. Cas12a-gFET leverages multiturnover trans-cleavage activity Cas12a intrinsic signal ultrasensitivity gFET. As demonstrated, achieves limit 1 aM ssDNA human papillomavirus 16 synthetic 10 dsDNA Escherichia coli plasmid without In addition, an array 48 sensors on single 1.5 cm × chip is employed improve data reliability. Finally, demonstrates capability discriminate single-nucleotide polymorphisms. Together, biosensor provides reliable, highly specific detections.

Language: Английский

SERS-based CRISPR/Cas assay on microfluidic paper analytical devices for supersensitive detection of pathogenic bacteria in foods DOI

Jianwen Zhuang,

Zhiying Zhao,

Kai Lian

et al.

Biosensors and Bioelectronics, Journal Year: 2022, Volume and Issue: 207, P. 114167 - 114167

Published: March 17, 2022

Language: Английский

Citations

195
Amplification-free CRISPR/Cas detection technology: challenges, strategies, and perspectives DOI
Huimin Li, Yi Xie, Fumin Chen

et al.

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

Citations

156
CRISPR‐Cas Biochemistry and CRISPR‐Based Molecular Diagnostics DOI Open Access

Zhengyan Weng,

Zheng You, Jie Yang

et al.

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

Citations

103
Paper-based sensors for bacteria detection DOI Open Access
Federico Mazur, Angie Davina Tjandra, Yingzhu Zhou

et al.

Nature Reviews Bioengineering, Journal Year: 2023, Volume and Issue: 1(3), P. 180 - 192

Published: Feb. 14, 2023

Language: Английский

Citations

100
A novel fluorescence biosensor based on CRISPR/Cas12a integrated MXenes for detecting Aflatoxin B1 DOI Creative Commons
Zhihui Wu, Da‐Wen Sun, Hongbin Pu

et al.

Talanta, Journal Year: 2022, Volume and Issue: 252, P. 123773 - 123773

Published: Aug. 6, 2022

Aflatoxin B1 (AFB1) contamination in food threatens global safety, and rapid quantitative detection of AFB1 remains a challenge. Herein, novel fluorescence biosensor was developed for based on CRISPR/Cas12a MXenes. Specifically, the well-designed activator locked by dual-AFB1 aptamers, Cas12a directly linked to crRNA form inactivated complexes, MXenes efficiently adsorbed FAM fluorophore-modified single-stranded DNA (ssDNA-FAM), quenching its fluorescence. In presence AFB1, released due preferential binding aptamer then activated trans-cleavage activity indiscriminately cleave ssDNA MXenes, leading recovery signal. The fluorescent had wide range from 0.001 80 ng mL-1, limit 0.92 pg ability detect within min. More importantly, platform demonstrates excellent performance real peanut samples.

Language: Английский

Citations

92
SERS-based microdevices for use as in vitro diagnostic biosensors DOI
Sungwoon Lee,

Hajun Dang,

Joung‐Il Moon

et al.

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(11), P. 5394 - 5427

Published: Jan. 1, 2024

This review explores various microdevices developed for applying SERS technology to in vitro diagnostics and delves into their clinical applications.

Language: Английский

Citations

53
Nucleic acid-assisted CRISPR-Cas systems for advanced biosensing and bioimaging DOI
Siyu Chen, Bo Gong, Cong Zhu

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 159, P. 116931 - 116931

Published: Jan. 11, 2023

Language: Английский

Citations

42
Biomedical SERS – the current state and future trends DOI Creative Commons
Dana Cialla‐May, Alois Bonifacio, Thomas Bocklitz

et al.

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(18), P. 8957 - 8979

Published: Jan. 1, 2024

Surface enhanced Raman spectroscopy (SERS) is meeting the requirements in biomedical science being a highly sensitive and specific analytical tool. By employing portable systems combination with customized sample pre-treatment, point-of-care-testing (POCT) becomes feasible. Powerful SERS-active sensing surfaces high stability modification layers if required are available for testing application complex biological matrices such as body fluids, cells or tissues. This review summarizes current state collection pretreatment SERS detection protocols, schemes,

Language: Английский

Citations

39
Recent Trends and Impact of Localized Surface Plasmon Resonance (LSPR) and Surface-Enhanced Raman Spectroscopy (SERS) in Modern Analysis DOI Creative Commons
Bibhu Prasad Nanda, Priyanka Rani,

Priyanka Paul

et al.

Journal of Pharmaceutical Analysis, Journal Year: 2024, Volume and Issue: 14(11), P. 100959 - 100959

Published: Feb. 28, 2024

An optical biosensor is a specialized analytical device that utilizes the principles of optics and light in bimolecular processes. Localized surface plasmon resonance (LSPR) phenomenon realm nanophotonics occurs when metallic nanoparticles (NPs) or nanostructures interact with incident light. Conversely, surface-enhanced Raman spectroscopy (SERS) an influential technique based on scattering, wherein it amplifies signals molecules they are situated near specific specially designed nanostructures. A detailed exploration recent ground-breaking developments biosensors employing LSPR SERS technologies has been thoroughly discussed along their underlying working mechanisms. chip created, featuring high-density deposition gold (AuNPs) under varying ligand concentration reaction duration substrate. ordinary description, visual illustration, provided for concepts such as sensogram, refractive index shift, (SPR), evanescent field, Rayleigh well electromagnetic enhancement chemical enhancement. both have advantages disadvantages, but widely used some over LSPR, like specificity, high sensitivity, multiplexing, versatility different fields. This review confirms elucidates significance disease biomarker identification. play vital role detection various types cancer, cervical ovarian endometrial prostate colorectal brain tumors. proposed offers potential applications early diagnosis monitoring viral disease, bacterial infectious diseases, fungal diabetes, cardiac biosensing. provide new direction environmental monitoring, food safety, refining impurities from water samples, lead detection. The understanding these still limited challenging.

Language: Английский

Citations

36
Convergence of Surface-Enhanced Raman Scattering with Molecular Diagnostics: A Perspective on Future Directions DOI
Namhyun Choi, Sebastian Schlücker

ACS Nano, Journal Year: 2024, Volume and Issue: 18(8), P. 5998 - 6007

Published: Feb. 12, 2024

Molecular diagnostics (MD) is widely employed in multiple scientific disciplines, such as oncology, pathogen detection, forensic investigations, and the pharmaceutical industry. Techniques polymerase chain reaction (PCR) revolutionized rapid accurate identification of nucleic acids (DNA, RNA). More recently, CRISPR its CRISPR-associated protein (Cas) have been a ground-breaking discovery that latest revolution molecular biology, including MD. Surface-enhanced Raman scattering (SERS) very attractive alternative to fluorescence currently most used optical readout In this Perspective, milestones development MD, SERS-PCR, next-generation approaches Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK) DNA Endonuclease-Targeted Trans (DETECTR), are briefly summarized. Our perspective on future convergence SERS with MD focused SERS-based CRISPR/Cas (SERS-CRISPR) since we anticipate many promising applications rapidly emerging field. We predict major developments will exploit advantages real-time monitoring superior brightness, photostability, spectral multiplexing potential nanotags an automated workflow for assays under isothermal, amplification-free conditions.

Language: Английский

Citations

25