Amplification-Free Detection of Mycobacterium Tuberculosis Using CRISPR-Cas12a and Graphene Field-Effect Transistors

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

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Validation of methods for enriching and detecting SARS-CoV-2 RNA in wastewater

Environmental Chemistry Letters, Год журнала: 2025, Номер unknown

Опубликована: Май 15, 2025

Язык: Английский

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One‐Step RAA and CRISPR‐Cas13a Method for Detecting Influenza B Virus

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

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Development of Novel Multiplex Detection Platforms Based on Multiepitope Nanobody Pairs for Rapid Screening of Waterborne Pathogens

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

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Facilitating self-testing with a fast, accurate, and simplified shelf-stable colorimetric LAMP system for Mpox and SARS-CoV-2 detection

Talanta, Год журнала: 2024, Номер 283, С. 127119 - 127119

Опубликована: Ноя. 4, 2024

Язык: Английский

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Three-Way Junction Probe triggered CRISPR/Cas14a1 enhanced EXPonential Amplification Reaction for Sensitive Pseudomonas aeruginosa Detection

Analytical Methods, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 13, 2024

Pseudomonas aeruginosa ( P. , PA) is a rod-shaped Gram-negative opportunistic bacterium capable of causing nosocomial infections, such as burn wound infections and meningitis.

Язык: Английский

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