Progress in molecular biology and translational science, Journal Year: 2024, Volume and Issue: unknown, P. 185 - 209
Published: Jan. 1, 2024
Language: Английский
Biosensors and Bioelectronics, Journal Year: 2024, Volume and Issue: unknown, P. 117092 - 117092
Published: Dec. 1, 2024
Language: Английский
Citations
8
ACS Nano, Journal Year: 2024, Volume and Issue: unknown
Published: Dec. 17, 2024
The simultaneous detection of proteins and microRNA (miRNA) at the single extracellular vesicle (EV) level shows great promise for precise disease profiling, owing to heterogeneity scarcity tumor-derived EVs. However, a highly reliable method multiple-target analysis EVs remains be developed. In this study, digital dual CRISPR-Cas-powered Single EV Evaluation (ddSEE) system was proposed enable concurrent surface protein inner miRNA single-molecule level. By optimizing reaction conditions CRISPR-Cas12a CRISPR-Cas13a, detected by Cas12a using antibody-DNA conjugates transfer signal DNA, while analyzed Cas13a through EV-liposome fusion. A microfluidic chip containing 188,000 microwells used convert CRISPR-Cas into assay format absolute quantification miRNA/protein-positive without bias fluorescence imaging, which can detect as few 214 EVs/μL. Finally, total 31 blood samples, 21 from breast cancer patients 10 healthy donors, were collected tested, achieving diagnostic accuracy 92% in distinguishing with donors. With its quantification, ease use, multiplexed capability, ddSEE demonstrates potential both research clinical applications.
Language: Английский
Citations
6
Critical Reviews in Biotechnology, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 28
Published: Sept. 22, 2024
CRISPR-based diagnostics (CRISPR/Dx) have revolutionized the field of molecular diagnostics. It enables home self-test, field-deployable, and point-of-care testing (POCT). Despite great potential CRISPR/Dx in diagnoses biologically complex diseases, preamplification template often is required for sensitive detection low-abundance nucleic acids. Various amplification-free systems were recently developed to enhance signal at sufficient sensitivity. Broadly, these are classified into five groups depending on enhancement strategies employed: CRISPR/Cas12a and/or CRISPR/Cas13a integrated with: (1) other catalytic enzymes (Cas14a, Csm6, Argonaute, duplex-specific nuclease, nanozyme, or T7 exonuclease), (2) rational-designed oligonucleotides (multivalent aptamer, tetrahedral DNA framework, RNA G-quadruplexes, roller machine, switchable-caged guide RNA, hybrid locked RNA/DNA probe, hybridized cascade "U" rich stem-loop RNA), (3) nanomaterials (nanophotonic structure, gold nanoparticle, micromotor, microbeads), (4) electrochemical piezoelectric plate biosensors (SERS nanoprobes, graphene field-effect transistor, redox primer exchange reaction), (5) cutting-edge technology platforms (digital bioanalysis, droplet microfluidic, smartphone camera, single nanoparticle counting). Herein, we critically discuss advances, pitfalls future perspectives acids detection. The continued refinement will pave road rapid, cost-effective, ultrasensitive, ultraspecific on-site without resorting target amplification, with ultimate goal establishing as paragon
Language: Английский
Citations
4
Published: Jan. 1, 2025
Language: Английский
Citations
0
Talanta, Journal Year: 2025, Volume and Issue: 289, P. 127772 - 127772
Published: Feb. 22, 2025
Language: Английский
Citations
0
ACS Sensors, Journal Year: 2025, Volume and Issue: unknown
Published: March 19, 2025
With the goal of impacting patient quality life and outcomes, sensor science offers significant potential to revolutionize healthcare by providing advances in detection molecular biomarkers for personalized clinical technologies. The community has achieved technical advancements that can impact diagnostics, health monitoring, disease treatment; however, many innovations remain confined laboratory, failing bridge translational gap between research real-world applications. This perspective presents a new direction community, where development centers on needs experiences primary beneficiaries: patients. We provide guidelines resources researchers engage with patients early continuously throughout process inform specifications better align technologies needs, improving their adoption impact. also present examples implementing patient-centered approach planning engagement research. In design impactful sensors patients, must expand focus beyond embrace approach, which will likely lead opportunities collaboration evolution community.
Language: Английский
Citations
0
Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown
Published: April 8, 2025
The CRISPR/Cas12a system is an emerging enzymatic tool for the development of enzyme-linked immunosorbent assay (ELISA) methods, owing to its robust signal amplification capability. Currently, most CRISPR/Cas12a-based ELISA approaches rely on strategies that convert target detection into nucleic acid analysis. This report presents a novel cascade reaction transduction and in method, utilizing β-galactosidase (β-gal)-mediated activation system. Carbendazim (CBD), widely used versatile broad-spectrum benzimidazole fungicide, was chosen as model analyte. In absence CBD, streptavidin-labeled β-gal captured by biotinylated secondary antibody immobilized microplate. catalyzes hydrolysis p-aminophenyl β-D-galactopyranoside generate p-aminophenol. compound subsequently facilitates decomposition MnO2 nanosheets, leading generation Mn2+ ions. ions modulate activity system, thus producing high fluorescence solution. presence amount microplate reduced, thereby preventing effective cleavage reporter molecule Cas12a, which results low signal. After systematically optimizing experimental conditions, developed method successfully detected demonstrating sensitivity, selectivity, applicability complex food matrices. comparison traditional acid-activated our approach, integrates β-gal-mediated, Mn2+-activated reactions ELISA, exhibits superior analytical performance, broadening sensitive convenient small-molecule
Language: Английский
Citations
0
Biosensors and Bioelectronics, Journal Year: 2025, Volume and Issue: 283, P. 117521 - 117521
Published: April 28, 2025
Language: Английский
Citations
0
Advanced Healthcare Materials, Journal Year: 2024, Volume and Issue: unknown
Published: Sept. 10, 2024
Abstract In the rapidly evolving field of life sciences and biomedicine, detecting low‐abundance biomolecules, ultraweak biosignals presents significant challenges. This has spurred a rapid development analytical techniques aiming for increased sensitivity specificity. These advancements, including signal amplification strategies integration biorecognition events, mark transformative era in bioanalytical precision accuracy. A prominent method among these innovations is immuno‐rolling circle (immuno‐RCA) technology, which effectively combines immunoassays with via RCA. process starts when targeted biomolecule, such as protein or cell, binds to an immobilized antibody probe on substrate. The introduction circular DNA template triggers RCA, leading exponential significantly enhanced intensity, thus target molecule detectable quantifiable even at single‐molecule level. review provides overview biosensing strategy extensive practical applications immuno‐RCA biomarkers. Furthermore, it scrutinizes limitations inherent sensors sets forth expectations their future trajectory. serves valuable reference advancing various domains, diagnostics, biomarker discovery, molecular imaging.
Language: Английский
Citations
1
Progress in molecular biology and translational science, Journal Year: 2024, Volume and Issue: unknown, P. 185 - 209
Published: Jan. 1, 2024
Language: Английский
Citations
0