CrRNA Conformation-Engineered CRISPR-Cas12a System for Robust and Ultrasensitive Nucleic Acid Detection

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 6, 2025

Despite the widespread application of CRISPR-Cas12a system in vitro diagnostics due to its high programmability and distinctive trans-cleavage activity, susceptibility crRNA component degradation sensitivity storage working conditions poses a significant challenge improving practical efficacy these diagnostic systems. Here, we show that engineered with covalently closed circular structure (C-crRNA) can replace traditional linear form functional complexes Cas12a protein, significantly enhancing anti-interference ability while maintaining specificity. Based on this finding, crRNA-mediated CRISPR molecular (CRCD) toolkit is developed successfully integrated standard nucleic acid amplification technique detect synthesized Human Papillomavirus type 16 (HPV-16) plasmids down 10 aM levels. Furthermore, CRCD applied for ultrasensitive detection 40 HPV-16 influenza A viruses clinical samples, results consistent those from PANTHER quantitative real-time polymerase chain reaction (qRT-PCR). In conclusion, strategy introduces novel paradigm engineering program Cas12a, which has potential revolutionize use CRISPR-based diagnostics.

Language: Английский

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A Review on Optical Biosensors for Monitoring of Uric Acid and Blood Glucose Using Portable POCT Devices: Status, Challenges, and Future Horizons

Biosensors, Journal Year: 2025, Volume and Issue: 15(4), P. 222 - 222

Published: March 31, 2025

The growing demand for real-time, non-invasive, and cost-effective health monitoring has driven significant advancements in portable point-of-care testing (POCT) devices. Among these, optical biosensors have emerged as promising tools the detection of critical biomarkers such uric acid (UA) blood glucose. Different transduction methods, like fluorescence, surface plasmon resonance (SPR), colorimetric approaches, are talked about, with a focus on how sensitive, specific, they are. Despite considerable advancements, several challenges persist, including sensor stability, miniaturization, interference effects, need calibration-free operation. This review also explores issues related to cost-effectiveness, data integration, wireless connectivity remote monitoring. further examines regulatory considerations commercialization aspects biosensors, addressing gap between research developments clinical implementation. Future perspectives emphasize integration artificial intelligence (AI) healthcare improved diagnostics, alongside development wearable implantable continuous Innovative potential change way people manage their by quickly accurately measuring glucose levels. is especially true decentralized solutions grows. By critically evaluating existing work exploring limitations opportunities field, this will help guide more efficient, accessible, reliable POCT devices that can improve patient outcomes quality life.

Language: Английский

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Multifunctional Fe3O4@CuS nanoparticle-driven colorimetric and photothermal immunochromatographic test strip for the sensitive detection of Salmonella typhimurium in milk

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1323, P. 343091 - 343091

Published: Aug. 10, 2024

Language: Английский

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Rapid and sensitive detection of exosomal microRNAs by terahertz metamaterials

Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy, Journal Year: 2025, Volume and Issue: 330, P. 125745 - 125745

Published: Jan. 17, 2025

Exosomal microRNAs (miRNAs) are among the most common biomarkers for tumor diagnosis. However, single-miRNA detection lacks ideal sensitivity and specificity diagnosing a certain in clinics. In this work, we fabricated convenient multi-miRNA platform sensitive specific on exosomal miRNAs plasma of patients using terahertz (THz) metamaterial biosensor basis strand displacement amplification (SDA) AuNPs. The proposed was highly to miRNA-15, miRNA-21, miRNA-145, miRNA-155, miRNA-423, miRNA-451, limit (LOD) obtained were 12.54 aM, 19.66 17.50 25.40 24.11 aM 26.59 respectively. constructed can be used diagnose pancreatic cancer effectively complex clinical samples. use multiple demonstrated strong practicality combined diagnosis with cancer. These studies demonstrate that multi-RNA boasts advantages such as low cost, rapid, high sensitivity, specificity, offering potential tool future applications.

Language: Английский

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Label-free electrochemical aptasensor for digoxin therapeutic monitoring in clinical samples via catalytic hairpin self-assembly and eATRP mediated signal amplification

Microchemical Journal, Journal Year: 2025, Volume and Issue: 211, P. 113047 - 113047

Published: Feb. 17, 2025

Language: Английский

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Two-dimensional nanozyme nanoarchitectonics customized electrochemical bio diagnostics and lab-on-chip devices for biomarker detection

Advances in Colloid and Interface Science, Journal Year: 2025, Volume and Issue: 341, P. 103474 - 103474

Published: March 20, 2025

Language: Английский

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RNA-based diagnostic innovations: A new frontier in diabetes diagnosis and management

Diabetes and Vascular Disease Research, Journal Year: 2025, Volume and Issue: 22(2)

Published: March 1, 2025

Background/Objective: Diabetes mellitus (DM) remains a major global health challenge due to its chronic nature and associated complications. Traditional diagnostic approaches, though effective, often lack the sensitivity required for early-stage detection. Recent advancements in molecular biology have identified RNA molecules, particularly non-coding RNAs such as microRNAs (miRNAs), long (lncRNAs), circular (circRNAs), promising biomarkers diabetes. This review aims explore role of RNA-based diagnosis, prognosis, management diabetes, highlighting their potential revolutionize diabetes care. Method: A comprehensive literature was conducted using electronic databases including PubMed, Scopus, Web Science. Articles published up 2024 were screened analyzed extract relevant findings related diagnostics Emphasis placed on studies demonstrating clinical utility, mechanistic insights, translational molecules. Results: Numerous species, miRNAs miR-375, miR-29, lncRNAs like H19 MEG3, exhibit altered expression patterns diabetic patients. These molecules are involved key regulatory pathways glucose metabolism, insulin resistance, β-cell function. Circulating detectable various biofluids, enabling non-invasive approaches. Emerging technologies, sequencing liquid biopsy platforms, enhanced specificity detection, fostering development novel tools personalized therapeutic strategies. Conclusion: hold significant promise advancing early risk stratification, monitoring Despite current challenges standardization validation, integration into routine practice could transform management, paving way precision medicine Further research multi-center trials essential validate these facilitate approval implementation.

Language: Английский

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Fast Screening of Urinary Tract Infections Using a Nitrite-Responsive Smartphone Fluorescent Sensor

Sensors and Actuators B Chemical, Journal Year: 2025, Volume and Issue: unknown, P. 137869 - 137869

Published: May 1, 2025

Language: Английский

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Rapid visual detection of Helicobacter pylori and vacA subtypes by Dual-Target RAA-LFD assay

Clinica Chimica Acta, Journal Year: 2024, Volume and Issue: 564, P. 119927 - 119927

Published: Aug. 15, 2024

Helicobacter pylori (H. pylori) infects over 50% of the global population and is a significant risk factor for gastric cancer. The pathogenicity H. primarily attributed to virulence factors such as vacA. Timely accurate identification, along with genotyping genes, are essential effective clinical management controlling its prevalence. In this study, we developed dual-target RAA-LFD assay rapid, visual detection genes (16s rRNA, ureA, vacA m1/m2), using recombinase aided amplification (RAA) combined lateral flow dipstick (LFD) methods. Both 16s rRNA ureA were selected identification ensure reliable accuracy. A was achieve at stable 37 °C within 20 min, followed by visualization (LFD). whole process, from results, took less than 30 min. 95 % limit (LOD) 16 s m1, m2 determined 3.8 × 10-2 ng/μL, 5.8 ng/μL 1.4 respectively. No cross-reaction observed in common pathogens including Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, showing assay's high specificity. evaluation performance assay. total 44 juice samples analyzed, immunofluorescence staining (IFS) quantitative polymerase chain reaction (qPCR) used reference results showed complete agreement qPCR findings, accurately identifying infection confirmed IFS 10 out patients. Furthermore, successfully genotyped m1/m2 among positive samples, achieving kappa (κ) value 1.00. study provides rapid method detecting minimizing dependency on sophisticated laboratory equipment specialized personnel. Clinical validation confirms efficacy promising tool effectively control prevalence aiding precise treatment pylori-associated diseases.

Language: Английский

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Smartphone-based mobile digital pressure sensor for quantitative point-of-care testing of respiratory syncytial virus infection

Talanta, Journal Year: 2025, Volume and Issue: 286, P. 127513 - 127513

Published: Jan. 4, 2025

Language: Английский

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