Microchemical Journal, Год журнала: 2024, Номер unknown, С. 112483 - 112483
Опубликована: Дек. 1, 2024
Язык: Английский
Nanoscale, Год журнала: 2025, Номер unknown
Опубликована: Янв. 1, 2025
Current molecular tests for tuberculosis (TB), such as whole genome sequencing and Xpert Mycobacterium tuberculosis/rifampicin resistance assay, exhibit limited sensitivity necessitate the pre-amplification step of target DNA. This limitation greatly increases detection time poses an increased risk infection. Here, we present a graphene field-effect transistor (GFET) based on CRISPR/Cas system detecting tuberculosis. The CRISPR/Cas12a has ability to specifically recognize cleave By integrating onto FET platform utilizing its electrical amplification capability, achieve rapid sensitive without requiring sample pre-amplification, with limit (LoD) low 2.42 × 10-18 M. Cas12a-GFET devices can differentiate 30 positive cases from 56 serum samples within 5 minutes. These findings highlight immense potential in future biological analysis clinical diagnosis.
Язык: Английский
Процитировано
2
Nature Communications, Год журнала: 2025, Номер 16(1)
Опубликована: Янв. 30, 2025
The CRISPR-based detection methods have been widely applied, yet they remain limited by the non-universal nature of one-pot diagnostic approaches. Here, we report a universal fluorescent method for epidemic pathogens, delivering results within 15-20 min. This uses heparin sodium to precisely tunes cis-cleavage capability Cas12 via interference with Cas12a-crRNA binding process, thereby generating significant fluorescence due accumulation isothermal amplification products. Additionally, this assay accommodates both classic and suboptimal PAMs, as well various Cas12a subtypes such LbCas12a, AsCas12a, AapCas12b. Such robust demonstrates sensitivity specificity exceeding 95% in monkeypox pseudovirus, influenza A virus, SARS-CoV-2 from saliva or wastewater samples, when compared qPCR RT-qPCR. Moreover, cost per thousand is $0.01 $0.04 only. Collectively, fast approach based on offers potential possibilities point-of-care testing.
Язык: Английский
Процитировано
2
Talanta, Год журнала: 2025, Номер 291, С. 127852 - 127852
Опубликована: Март 6, 2025
Язык: Английский
Процитировано
1
Trends in biotechnology, Год журнала: 2024, Номер 42(11), С. 1410 - 1426
Опубликована: Июль 20, 2024
Язык: Английский
Процитировано
7
Biosensors and Bioelectronics, Год журнала: 2024, Номер 263, С. 116617 - 116617
Опубликована: Июль 30, 2024
Язык: Английский
Процитировано
6
Опубликована: Янв. 1, 2025
Язык: Английский
Процитировано
0
Frontiers in Plant Science, Год журнала: 2025, Номер 16
Опубликована: Март 6, 2025
CRISPR-based disease detection has the potential to profoundly change how pathogens are detected in plant materials. However, there been a lack of research directed into improving explicitly CRISPR components that define these assays. To fill this technology gap, we have designed and optimized our CRISPR-Cas12a based platform by showcasing its capability detecting pathogen group rising importance, Candidatus Phytoplasma. Most assays utilize isothermal pre-amplification steps, which may boost sensitivity yet often lead false positives. Aiming for pre-amplification-free assay maintain accuracy, screened multiple Cas12a orthologs variants found LbCas12a-Ultra be most sensitive Cas12a. We further improved system using stem-loop reporters various sizes 7nt significantly outperformed other as well commonly used linear reporters. When reporter was combined with best-performing LbCas12a-Ultra, 10-fold increase over standard LbCas12a assay. enhance coverage highly diverse phytoplasmas, tested multiplex method predicted target nearly 100% all documented phytoplasma species on NCBI. A lateral flow also developed accommodate instrument-free reagents. Our study demonstrates an wide applications can easily integrated almost any Cas12a-based boosted sensitivity.
Язык: Английский
Процитировано
0
Microbial Biotechnology, Год журнала: 2025, Номер 18(4)
Опубликована: Апрель 1, 2025
ABSTRACT We developed a sensitive and specific method based on recombinase‐aided amplification (RAA) clustered regularly interspaced short palindromic repeats (CRISPR)‐CRISPR‐associated protein 13a (Cas13a). This method, named CRISPR‐based Rapid Efficient Test (CRISPRET), is designed for the early diagnosis of Influenza B (FluB) with aim shortening its transmission chain. identified conserved regions in Virus (IBV) NS gene forward reverse primers along crRNAs. then established optimised reaction system, Nucleic Acid Positive Reference Materials IBV were used to evaluate detection limit (DL) CRISPRET. Additionally, we collected 257 clinical samples, comprising 127 samples from patients infection 130 healthy individuals, subjected them dual using CRISPRET qPCR positive predictive value (PPV), negative (NPV), sensitivity specificity one primer, two primers, crRNAs establish optimise CRISPR ET. The demonstrated DL 500 copies·μL −1 when assisted by appropriate equipment. Despite requiring auxiliary equipment 30‐min reaction, ET enables nucleic acid within approximately first 5 min, achieving high (100%), (97.69%), PPV (97.69%) NPV concordance rate 98.83% qPCR. offers simple, field‐applicable, one‐step rapid IBV. It has strong potential field‐testing applications intelligent integration into existing diagnostic systems.
Язык: Английский
Процитировано
0
Analytical Chemistry, Год журнала: 2025, Номер unknown
Опубликована: Май 16, 2025
The rapid and multiplexed detection of waterborne viruses is crucial for infection prevention. However, current methods are limited by low-quality probes, instrument dependency, time-consuming procedures. In this study, we developed a high-performance, nanobody pair-based, multichannel homogeneous platform the simultaneous three viruses─SARS-CoV-2, norovirus, influenza A virus─in aquatic environments. To identify robust sensitive multiepitope pairs these viruses, utilized pressure-assisted screening docking techniques. For detection, synthesized distinct SiO2@TQD types with unique excitation wavelengths, each acting as an independent signal label. By integrating antibody arrays labels into unified platform, sensor capable detecting all within 30 min. system demonstrated limits low 1.56 pg/mL SARS-CoV-2 antigen, 0.1 norovirus 0.39 virus surpassing conventional antigen kits sensitivity enhancement 160.26-6.25 × 104-fold. Notable advantages include exceptional specificity, accuracy, stability. This work not only provides transformative solution monitoring pathogens but also establishes versatile framework developing platforms other infectious agents.
Язык: Английский
Процитировано
0
International Journal of Biological Macromolecules, Год журнала: 2025, Номер 314, С. 144400 - 144400
Опубликована: Май 19, 2025
Язык: Английский
Процитировано
0