Detection of antibiotic-resistance genes in bacterial pathogens using a Cas12a/3D DNAzyme colorimetric paper sensor DOI Creative Commons
Hua Gao, Yanan Li, Yaqiong Li

et al.

Fundamental Research, Journal Year: 2023, Volume and Issue: unknown

Published: May 1, 2023

The rapid detection of antibiotic-resistant genes in bacterial pathogens is critical combating global health crises. Herein, we report a CRISPR/Cas12a-based colorimetric paper sensor, where the trans-cleavage activity Cas12a was post-amplified by rolling circle replication, resulting generation 3D DNAzyme. DNAzyme adhered strongly to surface, creating highly bioactive sensor containing high densities functional DNAzymes. This assay effective for gene, NDM-1, with sensitivity. In absence NDM-1 catalyzed reaction, blue-colored signal while presence collateral cleavage activated, leading template, thus preventing and producing no signal. provides low-cost carried various pathogenic microorganisms femtomolar-level sensitivity results that are visible naked eye. entire analysis requires less than 90 minutes time. Due programmable design CRISPR probe, platform has significant potential quick responses new epidemics.

Language: Английский

Nucleic Acid Enzyme‐Activated CRISPR‐Cas12a With Circular CRISPR RNA for Biosensing DOI Open Access
Yunping Wu, Dingran Chang, Yangyang Chang

et al.

Small, Journal Year: 2023, Volume and Issue: 19(41)

Published: June 9, 2023

clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems are increasingly used in biosensor development. However, directly translating recognition events for non-nucleic acid targets by CRISPR into effective measurable signals represents an important ongoing challenge. Herein, it is hypothesized and confirmed that RNAs (crRNAs) a circular topology efficiently render Cas12a incapable of both site-specific double-stranded DNA cutting nonspecific single-stranded trans cleavage. Importantly, shown nucleic enzymes (NAzymes) with RNA-cleaving activity can linearize the crRNAs, activating CRISPR-Cas12a functions. Using ligand-responsive ribozymes DNAzymes as molecular elements, demonstrated target-triggered linearization crRNAs offers great versatility biosensing. This strategy termed "NAzyme-Activated Circular RNA (NA3C)." Use NA3C clinical evaluation urinary tract infections using Escherichia coli-responsive DNAzyme to test 40 patient urine samples, providing diagnostic sensitivity 100% specificity 90%, further demonstrated.

Language: Английский

Citations

26
SATCAS: A CRISPR/Cas13a-based simultaneous amplification and testing platform for one-pot RNA detection and SNPs distinguish in clinical diagnosis DOI Creative Commons
Ting Wang,

Linlin Bai,

Guoling Wang

et al.

Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: 263, P. 116636 - 116636

Published: Aug. 5, 2024

The clinical diagnosis of pathogen infectious diseases increasingly requires sensitive and rapid RNA detection technologies. RNA-guided clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system has shown immense potential in molecular diagnostics due to its trans-cleavage activity. However, most Cas13a-based methods require an amplicon transcription step, the multi-step open-tube operations are prone contamination, limiting their widespread application. Here, we propose ultrasensitive (single-copy range, ∼aM) (within 40 min) isothermal one-pot platform, termed SATCAS (Simultaneous Amplification Testing platform based on Cas13a). This method effectively distinguishes viable bacteria (0%-100%) under constant total bacterial conditions, demonstrating robustness universality. excels identifying single nucleotide polymorphisms (SNPs), particularly detecting 0.5% drug-resistant mutations. We validated by infections biological samples from 68 HBV, 23 EBV, 48 SARS-CoV-2 patients, achieving 100% sensitivity, 92.86% specificity, 97.06% accuracy HBV infection testing. anticipate that broad application early diagnosis, subtyping, drug resistance detection, point-of-care monitoring diseases.

Language: Английский

Citations

11
CRISPR-based nucleic acid assays for food authentication DOI
Ruijie Deng, Lin Xu, Yong Zhang

et al.

Trends in Food Science & Technology, Journal Year: 2024, Volume and Issue: 145, P. 104351 - 104351

Published: Jan. 28, 2024

Language: Английский

Citations

9
NAPTUNE: nucleic acids and protein biomarkers testing via ultra-sensitive nucleases escalation DOI Creative Commons
Tao Hu,

Xinxin Ke,

Yingying Yu

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Feb. 4, 2025

Language: Английский

Citations

2
Dual-mode colorimetric and fluorescence biosensors for the detection of foodborne bacteria DOI
Raed Obaid Saleh, Yasir Qasim Almajidi, Sofiène Mansouri

et al.

Clinica Chimica Acta, Journal Year: 2023, Volume and Issue: 553, P. 117741 - 117741

Published: Dec. 27, 2023

Language: Английский

Citations

17
A portable CRISPR-Cas12a triggered photothermal biosensor for sensitive and visual detection of Staphylococcus aureus and Listeria monocytogenes DOI

Xijuan Gu,

Qu Tang,

Xiaoxia Kang

et al.

Talanta, Journal Year: 2024, Volume and Issue: 271, P. 125678 - 125678

Published: Jan. 21, 2024

Language: Английский

Citations

8
Clostridium butyricum Argonaute-powered programmable platform using a magnetic nanoparticle tetrahedral DNA encoding system for the ultrasensitive and multiplexed detection of non-nucleic acid targets DOI
Letian Li, Mengjiao Wang, Junping Wen

et al.

Chemical Engineering Journal, Journal Year: 2024, Volume and Issue: 484, P. 149548 - 149548

Published: Feb. 15, 2024

Language: Английский

Citations

8
Rapid and sensitive detection of two fungal pathogens in soybeans using the recombinase polymerase amplification/CRISPR‐Cas12a method for potential on‐site disease diagnosis DOI

Xiwen Sun,

Rong Lei, Haipeng Zhang

et al.

Pest Management Science, Journal Year: 2023, Volume and Issue: 80(3), P. 1168 - 1181

Published: Oct. 24, 2023

Abstract BACKGROUND Diaporthe aspalathi and caulivora are two of the fungal pathogens causing soybean stem canker (SSC) in soybean, which is one most widespread diseases growing regions can cause 100% loss yield. Current methods for detection pathogens, including morphological identification molecular detection, mostly limited by need professional laboratories staff. To develop a method potential on‐site diagnosis SSC, we designed rapid assay combining recombinase polymerase amplification (RPA) CRISPR‐Cas12a‐based diagnostics to specifically detect D. caulivora. RESULTS The translation elongation factor 1‐alpha gene was employed as target evaluate specificity sensitivity this assay. RPA/CRISPR‐Cas12a system has excellent distinguish from closely related species. sensitivities RPA/CRISPR‐Cas12a‐based fluorescence lateral flow 14.5 copies 24.6 copies, respectively. This hyphae inoculated stems at 12 days after inoculation recovery high 86% hyphae‐spiked seed powder. total time DNA extraction not more than 60 min. CONCLUSION developed plant includes with magnetic beads or extraction, isothermal nucleic acid 39 °C, CRISPR‐Cas12a cleavage reaction 37 endpoint visualization room temperature. RPA reagents be preloaded microcentrifuge tube simplify procedures field. Both realized on portable incubator, results visualized using strips flashlight. requires minimal equipment operator training, promising applications disease screening, port inspection, controlling pathogen transmission crop. © 2023 Society Chemical Industry.

Language: Английский

Citations

15
Advances in sensor developments for cell culture monitoring DOI Creative Commons
Ka Ram Kim, Woon‐Hong Yeo

BMEMat, Journal Year: 2023, Volume and Issue: 1(4)

Published: Sept. 19, 2023

Abstract Cell culture encompasses procedures for extracting cells from their natural tissue and cultivating them under controlled artificial conditions. During this process, various factors, including cell physiological/morphological properties, environments, metabolites, contaminants, have to be precisely monitored the survival of pursuit desired properties cells. This review summarizes recent advances in sensor technologies manufacturing strategies platforms using traditional plastics, microfluidic chips, scalable bioreactors. We share details newly developed biological sensors, chemical optical electronic chip technologies, material integration methods. The precise control parameters based on feedback by these sensors electronics enhances quality throughput.

Language: Английский

Citations

14
In Situ Cas12a-Based Allele-Specific PCR for Imaging Single-Nucleotide Variations in Foodborne Pathogenic Bacteria DOI
Xinmiao Liu, Hao Yang, Jun O. Liu

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(5), P. 2032 - 2040

Published: Jan. 26, 2024

In situ profiling of single-nucleotide variations (SNVs) can elucidate drug-resistant genotypes with single-cell resolution. The capacity to directly "see" genetic information is crucial for investigating the relationship between mutated genes and phenotypes. Fluorescence in hybridization serves as a canonical tool imaging; however, it cannot detect subtle sequence alteration including SNVs. Herein, we develop an Cas12a-based amplification refractory mutation system-PCR (ARMS-PCR) method that allows visualization SNVs related quinolone resistance inside cells. discriminating enhanced by incorporating optimized mismatched bases allele-specific primers, thus allowing specifically amplify quinolone-resistant genes. After ARMS-PCR, employed modified Cas12a/CRISPR RNA tag amplicon, thereby enabling specific binding fluorophore-labeled DNA probes. precisely quantify Salmonella enterica bacterial mixture. Utilizing this method, investigated survival competition quinolone-sensitive bacteria toward antimicrobial peptides indicated enrichment under colistin sulfate stress. ARMS-PCR holds potential cellular phenotypes gene regulation resolution at level.

Language: Английский

Citations

5