Microchimica Acta, Journal Year: 2024, Volume and Issue: 192(1)
Published: Dec. 21, 2024
Language: Английский
Nanoscale, Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 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.
Language: Английский
Citations
2
Talanta, Journal Year: 2025, Volume and Issue: 291, P. 127852 - 127852
Published: March 6, 2025
Language: Английский
Citations
1
Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: 263, P. 116617 - 116617
Published: July 30, 2024
Language: Английский
Citations
6
Trends in biotechnology, Journal Year: 2024, Volume and Issue: 42(11), P. 1410 - 1426
Published: July 20, 2024
Language: Английский
Citations
5
Published: Jan. 1, 2025
Language: Английский
Citations
0
Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)
Published: Jan. 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.
Language: Английский
Citations
0
Frontiers in Plant Science, Journal Year: 2025, Volume and Issue: 16
Published: March 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.
Language: Английский
Citations
0
International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: 314, P. 144400 - 144400
Published: May 19, 2025
Language: Английский
Citations
0
Viruses, Journal Year: 2025, Volume and Issue: 17(5), P. 721 - 721
Published: May 17, 2025
Foot and mouth disease virus (FMDV) is a highly pathogenic that mainly infects cloven hooved animals, such as pigs. The establishment of rapid, sensitive accurate point-of-care detection method critical for the timely identification elimination infected pigs controlling this disease. In study, RT-RAA-CRISPR/Cas13a was developed FMDV serotype O in Six pairs RT-RAA primers were designed based on conserved gene sequence O, optimal amplification reaction temperatures screened. CRISPR-derived RNA (crRNA) further target band most efficient crRNA results revealed FMDV-O-F4/R4 primer set, temperature 37 °C. Moreover, crRNA4 exhibited strongest signal among six crRNAs. established demonstrated high specificity no cross-reactivity with other common swine pathogens Senecavirus A (SVA), porcine reproductive respiratory (PRRSV), epidemic diarrhea (PEDV), circovirus type 2 (PCV2), classical fever (CSFV), pseudorabies (PRV), additionally, it observed to be sensitive, limit 19.1 copies/µL. repeatability also good. This could produce stable fluorescence good when three independent experiments yielded same results. validation test using types simulated clinical samples (including swab, tissue, serum samples) 100% concordance rate. visualized via reader or lateral flow strips (LFSs). Thus, specific expected applied rapid situ.
Language: Английский
Citations
0
Biotechnology Advances, Journal Year: 2025, Volume and Issue: unknown, P. 108609 - 108609
Published: May 1, 2025
Language: Английский
Citations
0