Biosensing Intestinal Alkaline Phosphatase by Pregnancy Test Strips Based on Target-Triggered CRISPR-Cas12a Activity to Monitor Intestinal Inflammation DOI
Zhenzhen Jia, Yujie Zhang, Chuanyu Zhang

et al.

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(37), P. 14111 - 14118

Published: Sept. 5, 2023

With an increasing incidence worldwide, inflammatory bowel disease (IBD) is a chronic affecting the gastrointestinal tract, which impairs life quality of patients. Therefore, it great significance to construct sensitive, simple, and convenient biosensor analyze IBD-associated biomarkers for auxiliary diagnosis IBD. Intestinal alkaline phosphatase (IAP), expressed by intestinal epithelium, endogenous protein that thought play vital role in maintaining homeostasis considered potential biomarker Here, IAP detection method was developed using pregnancy test strips dephosphorylation. Initially, double-stranded DNA (dsDNA) designed respond acted as activator Cas12a. In presence IAP, dsDNA not digested lambda exonuclease (λ exo), hybridized Cas12a-crRNA duplex resulted activation trans-cleavage Further, activated Cas12a cleaved single-strand (ssDNA) linker MBs-ssDNA-hCG probe, triggering release hCG. magnetic separation, released hCG could be quantitatively detected strips. levels were analyzed feces from colitis healthy mice The results showed level (3.89 ± 1.92 U/L) much lower than (39.64 24.93 U/L), indicating correlation between inflammation. Taken together, user-friendly assay based on constructed monitor used diagnostic approach IBD clinical scene.

Language: Английский

On-Site Fluorescent Detection of Sepsis-Inducing Bacteria using a Graphene-Oxide CRISPR-Cas12a (GO-CRISPR) System DOI Creative Commons
Tom Kasputis, Yawen He, Qiaoqiao Ci

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(6), P. 2676 - 2683

Published: Jan. 30, 2024

Sepsis is an extremely dangerous medical condition that emanates from the body's response to a pre-existing infection. Early detection of sepsis-inducing bacterial infections can greatly enhance treatment process and potentially prevent onset sepsis. However, current point-of-care (POC) sensors are often complex costly or lack ideal sensitivity for effective detection. Therefore, it crucial develop rapid sensitive biosensors on-site bacteria. Herein, we developed graphene oxide CRISPR-Cas12a (GO-CRISPR) biosensor bacteria in human serum. In this strategy, single-stranded (ssDNA) FAM probes were quenched with single-layer (GO). Target-activated Cas12a trans-cleavage was utilized degradation ssDNA probes, detaching short GO recovering fluorescent signals. Under optimal conditions, employed our GO-CRISPR system Salmonella Typhimurium (S. Typhimurium) as low 3 × 103 CFU/mL serum, well good specificity toward other competing addition, exhibited excellent S. spiked The offers superior rapidity has potential early resource-limited settings, expediting patients at risk

Language: Английский

Citations

18
RT‐RPA Assisted CRISPR/Cas12a Based One‐Pot Rapid and Visual Detection of the Pan‐Dengue Virus DOI
Pooja Bhardwaj,

Preeti Dhangur,

Alagarasu Kalichamy

et al.

Journal of Medical Virology, Journal Year: 2025, Volume and Issue: 97(2)

Published: Feb. 1, 2025

ABSTRACT Globally ≤ 4 billion of the population are at potential risk contracting dengue virus (DENV) infection. Seasonal outbreaks frequently reported causing a high healthcare burden. Undiagnosed DENV can lead to severe morbidity and mortality. Early diagnosis relies on molecular methods, which impractical in resource‐constrained settings (RCSs). Dengue be caused by any four distinct serotypes. Therefore, simple method for rapid Pan‐DENV serotypes is utmost importance RCSs. A fluorescence detection platform using RT‐RPA CRISPR/Cas12a was developed targeting nonstructural 1 ( NS1 ) gene DENV‐1, 2, 3, envelope E DENV‐2. Further, crRNA specific were designed facilitate detection. Analytical sensitivity determined synthetic RNA genome. Clinical validation assay performed extracted from AES/AFI clinical samples. The CRISPR/Cas12a‐based detect all viz 1−4 single pot This showed limit ≥ 781 zg reaction − , 1.81 ag −1 62.5 fg 2.5 pg DENV‐2, DENV‐3, DENV‐4 template, respectively. Our demonstrated analytic 10 ng DENV‐1 DENV‐4, 0.5 DENV‐3 genomes. no cross‐reactivity with other related etiologies tested AFI/AES. With 76 samples (DENV PCR positive = 16, negative 60), 93.7% 100% specificity an overall accuracy 98.7% displayed comparable results that RT‐PCR. ease interpretation Pan‐DENV, represents as ideal point‐of‐care test. upon field‐deployment could help reducing burden, provide differential support initiating early prompt treatment patients RCS.

Language: Английский

Citations

2
CRISPR/Cas12a-mediated cyclic signal amplification and electrochemical reporting strategy for rapid and accurate sensing of Vibrio parahaemolyticus in aquatic foods DOI

Haoyang Xu,

Qi Chen,

Xianzhuo Meng

et al.

Biosensors and Bioelectronics, Journal Year: 2025, Volume and Issue: 277, P. 117284 - 117284

Published: Feb. 19, 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

58
Designing a CRISPR/Cas12a- and Au-Nanobeacon-Based Diagnostic Biosensor Enabling Direct, Rapid, and Sensitive miRNA Detection DOI

Tong Luo,

Jiacheng Li, Yao He

et al.

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(17), P. 6566 - 6573

Published: April 22, 2022

Direct, rapid, sensitive, and selective detection of nucleic acids in complex biological fluids is crucial for medical early diagnosis. We herein combine the trans-cleavage ability clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a with Au-nanobeacon to establish a CRISPR-based biosensor, providing rapid miRNA high speed attomolar sensitivity. In this strategy, we first report that activity CRISPR/cas12a, which was previously reported be triggered only by target ssDNA or dsDNA, can activated directly. Therefore, method direct, i.e., does not need conversion into its complementary DNA (cDNA). Meanwhile, as compared traditional reporters molecular beacon (MB) reporters, exhibit improved reaction kinetics assay, miRNA-21 could detected very sensitivity 5 min. Finally, proposed strategy enables determination samples, potential tool

Language: Английский

Citations

42
Portable biosensor for on-site detection of kanamycin in water samples based on CRISPR-Cas12a and an off-the-shelf glucometer DOI
Junhua Chen,

Gu Shi,

Chong Yan

et al.

The Science of The Total Environment, Journal Year: 2023, Volume and Issue: 872, P. 162279 - 162279

Published: Feb. 16, 2023

Language: Английский

Citations

30
Quantitative or digital PCR? A comparative analysis for choosing the optimal one for biosensing applications DOI
Haoqing Zhang, Lei Cao, Jan Brodský

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2024, Volume and Issue: 174, P. 117676 - 117676

Published: March 28, 2024

Language: Английский

Citations

14
Development of a Rapid and Sensitive RPA-CRISPR/Cas12a Assay for Non-invasive Pre-implantation Genetic Testing DOI

Yuqin Liu,

Linghan Zhang,

Weifu Lei

et al.

Analytica Chimica Acta, Journal Year: 2025, Volume and Issue: unknown, P. 343687 - 343687

Published: Jan. 1, 2025

Language: Английский

Citations

1
Accelerated CRISPR/Cas12a-based small molecule detection using bivalent aptamer DOI
Xiuping Li, Xiujin Chen,

Minxin Mao

et al.

Biosensors and Bioelectronics, Journal Year: 2022, Volume and Issue: 217, P. 114725 - 114725

Published: Sept. 16, 2022

Language: Английский

Citations

37
CRISPR/Cas12a-Triggered Chemiluminescence Enhancement Biosensor for Sensitive Detection of Nucleic Acids by Introducing a Tyramide Signal Amplification Strategy DOI
Tao Hu,

Xinxin Ke,

Yangjing Ou

et al.

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(23), P. 8506 - 8513

Published: May 30, 2022

CRISPR-based biosensors have attracted increasing attention in accurate and sensitive nucleic acid detection. In this work, we report a CRISPR/Cas12a-triggered chemiluminescence enhancement biosensor for the ultrasensitive detection of acids by introducing tyramide signal amplification first time (termed CRICED). The hybrid chain DNA (crDNA) formed NH2-capture (capDNA) biotin-recognition (recDNA) was preferentially attached to magnetic beads (MBs), streptavidin–HRP subsequently introduced obtain MB@HRP-crDNA. presence target, activated CRISPR/Cas12a is capable randomly cutting initiator (intDNA) into vast short products, thus fractured intDNA could not trigger toehold-mediated DNA-strand displacement reaction (TSDR) event with After addition tyramine–AP H2O2, abundant HRP–tyramine–AP emerges through covalent attachment HRP–tyramine, exhibiting enhanced (CL) signals or visual image readouts. By virtue biosensor, achieved high sensitivity synthetic target amplified plasmid using recombinase polymerase (RPA) as low 17 pM single-copy detection, respectively. Our proposed CRICED further evaluated test 20 HPV clinical samples, showing superior 87.50% specificity 100.00%. Consequently, platform be an attractive means imaging holds promising strategy practical application diagnostics.

Language: Английский

Citations

36