DNA-Assisted CRISPR-Cas12a Enhanced Fluorescent Assay for Protein Detection in Complicated Matrices DOI
Sathishkumar Munusamy, Haiyan Zheng,

Rana Jahani

et al.

ACS Applied Bio Materials, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 19, 2024

Proteins are important biological macromolecules that perform a wide variety of functions in the cell and human body, can serve as biomarkers for early diagnosis prognosis diseases well monitoring effectiveness disease treatment. Hence, sensitive accurate detection proteins biospecimens is imperative. However, at present, there no ideal method available clinical samples, many which present ultralow (less than 1 pM) concentrations complicated matrices. Herein, we report an ultrasensitive selective DNA-assisted CRISPR-Cas12a enhanced fluorescent assay (DACEA) protein with limits reaching low attomolar concentrations. The high sensitivity was accomplished through combined DNA barcode amplification (by using dual-functionalized AuNPs) CRISPR analysis, while selectivity resistance to matrix effects our were via formation protein-antibody sandwich structure specific recognition Cas12a (under guidance crRNA) toward designed target ssDNA. Given its ability accurately sensitively detect trace amounts matrices, DACEA platform pioneered this work has potential application routine biomarker testing.

Language: Английский

Innovative Diagnostic Approaches and Challenges in the Management of HIV: Bridging Basic Science and Clinical Practice DOI Creative Commons

Mohd Afzal,

Shagun Agarwal, Rabab Hassan Elshaikh

et al.

Life, Journal Year: 2025, Volume and Issue: 15(2), P. 209 - 209

Published: Jan. 30, 2025

Human Immunodeficiency Virus (HIV) remains a major public health challenge globally. Recent innovations in diagnostic technology have opened new pathways for early detection, ongoing monitoring, and more individualized patient care, yet significant barriers persist translating these advancements into clinical settings. This review highlights the cutting-edge methods emerging from basic science research, including molecular assays, biosensors, next-generation sequencing, discusses practical logistical challenges involved their implementation. By analyzing current trends techniques management strategies, we identify critical gaps propose integrative approaches to bridge divide between laboratory innovation effective application. work emphasizes need comprehensive education, supportive infrastructure, multi-disciplinary collaborations enhance utility of improving outcomes patients with HIV.

Language: Английский

Citations

2
CRISPR-Driven Biosensors: A New Frontier in Rapid and Accurate Disease Detection DOI

ZahraSadat Razavi,

M. Soltani, Mohammad Souri

et al.

Critical Reviews in Analytical Chemistry, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 25

Published: Sept. 17, 2024

This comprehensive review delves into the advancements and challenges in biosensing, with a strong emphasis on transformative potential of CRISPR technology for early rapid detection infectious diseases. It underscores versatility CRISPR/Cas systems, highlighting their ability to detect both nucleic acids non-nucleic acid targets, seamless integration isothermal amplification techniques. The provides thorough examination latest developments CRISPR-based biosensors, detailing unique properties such as high specificity programmability, which make them particularly effective detecting disease-associated acids. While focuses due its critical role diagnosing diseases, it also explores broader applications thereby acknowledging technology's potential. Additionally, identifies existing challenges, need improved signal real-world applicability, offers future perspectives aimed at overcoming these hurdles. ultimate goal is advance development highly sensitive specific biosensors that can be used widely improving human health, point-of-care settings resource-limited environments.

Language: Английский

Citations

14
Development of RT-RPA-PfAgo-mediated assay for rapid and sensitive detection of influenza B virus DOI
Weidong Qian, H. C. Zeng,

Xueting Xu

et al.

Microchemical Journal, Journal Year: 2025, Volume and Issue: unknown, P. 113138 - 113138

Published: Feb. 1, 2025

Language: Английский

Citations

1
CRISPR-based electrochemical biosensors: an alternative for point-of-care diagnostics? DOI
Dagwin Wachholz, Lauro T. Kubota

Talanta, Journal Year: 2024, Volume and Issue: 278, P. 126467 - 126467

Published: June 28, 2024

Language: Английский

Citations

8
Advances, challenges, and opportunities for food safety analysis in the isothermal nucleic acid amplification/CRISPR-Cas12a era DOI

Pengpeng Xue,

Yubo Peng, Renjing Wang

et al.

Critical Reviews in Food Science and Nutrition, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 16

Published: April 24, 2024

Global food safety stands out as a prominent public concern, affecting populations worldwide. The recurrent challenge of incidents reveals the need for robust inspection framework. In recent years, integration isothermal nucleic acid amplification with CRISPR-Cas12a techniques has emerged promising tool molecular detection hazards, presenting next generation biosensing detection. This paper provides comprehensive review current state research on synergistic application and technology in field safety. innovative combination not only enriches analytical tools, but also improving assay performance such sensitivity specificity, addressing limitations traditional methods. summarized various methodologies by diverse concerns, including pathogenic bacterium, viruses, mycotoxins, adulteration, genetically modified foods. Each section elucidates specific strategies employed highlights advantages conferred. Furthermore, discussed challenges faced this context safety, offering insightful discussions potential solutions future prospects.

Language: Английский

Citations

6
Harnessing the power of clustered regularly interspaced short palindromic repeats (CRISPR) based microfluidics for next-generation molecular diagnostics DOI
Rasanpreet Kaur, Saurabh Kumar Gupta, Arjun Chauhan

et al.

Molecular Biology Reports, Journal Year: 2024, Volume and Issue: 51(1)

Published: Aug. 8, 2024

Language: Английский

Citations

4
Leveraging Synthetic Antibody–DNA Conjugates to Expand the CRISPR-Cas12a Biosensing Toolbox DOI
Elisa Paialunga,

N. Bagheri,

Marianna Rossetti

et al.

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 2, 2025

We report here the use of antibody-DNA conjugates (Ab-DNA) to activate collateral cleavage activity CRISPR-Cas12a enzyme. Our findings demonstrate that Ab-DNA effectively trigger CRISPR-Cas12a, enabling transduction antibody-mediated recognition events into fluorescence outputs. developed two different immunoassays using an as activator Cas12a: CRISPR-based immunosensing assay (CIA) for detecting SARS-CoV-2 spike S protein, which shows superior sensitivity compared with traditional enzyme-linked immunosorbent (ELISA), and immunomagnetic (CIMA). Notably, CIMA successfully detected protein in undiluted saliva a limit detection (LOD) 890 pM 2 h assay. results underscore benefits integrating Cas12a-based signal amplification antibody methods. The potential conjugates, combined CRISPR technology, offers promising alternative conventional enzymes used could facilitate development versatile analytical platforms non-nucleic acid targets.

Language: Английский

Citations

0
Structure-switching G-quadruplex: An efficient CRISPR/Cas12a signal reporter for label-free colorimetric biosensing DOI
Yang Chen, Bei Lv, Wenzhu Wang

et al.

International Journal of Biological Macromolecules, Journal Year: 2025, Volume and Issue: 307, P. 142410 - 142410

Published: March 22, 2025

Language: Английский

Citations

0
Machine learning and statistical classification in CRISPR-Cas12a diagnostic assays DOI Creative Commons
Nathan Khosla, Jake M. Lesinski, Marcus Haywood-Alexander

et al.

Biosensors and Bioelectronics, Journal Year: 2025, Volume and Issue: 279, P. 117402 - 117402

Published: March 31, 2025

CRISPR-based diagnostics have gained increasing attention as biosensing tools able to address limitations in contemporary molecular diagnostic tests. To maximize the performance of assays, much effort has focused on optimizing chemistry and biology reaction. However, less been paid improving techniques used analyze data. date, decisions typically involve various forms slope-based classification. Such methods are superior traditional based assessing absolute signals, but still limitations. Herein, we establish benchmarks (total accuracy, sensitivity, specificity) using common methods. We compare these benchmark with three different quadratic empirical distribution function statistical tests, finding significant improvements speed accuracy when applied a clinical data set. Two techniques, Kolmogorov-Smirnov Anderson-Darling report lowest time-to-result highest total test accuracy. Furthermore, developed long short-term memory recurrent neural network classify CRISPR-biosensing data, achieving 100 % specificity our model Finally, provide guidelines choosing classification method parameters that best suit assay's needs.

Language: Английский

Citations

0
MAIGRET: a CRISPR-based immunoassay that employs antibody-induced cell-free transcription of CRISPR guide RNA strands DOI Creative Commons
Francesca Miceli,

Sara Bracaglia,

Daniela Sorrentino

et al.

Nucleic Acids Research, Journal Year: 2025, Volume and Issue: 53(6)

Published: March 20, 2025

Abstract Here we report on the development of a CRISPR-based assay for sensitive and specific detection antibodies antigens directly in complex sample matrices. The assay, called Molecular Assay based antibody-Induced Guide-RNA Enzymatic Transcription (MAIGRET), is use responsive synthetic DNA template that triggers cell-free vitro transcription guide RNA strand upon recognition target antibody. Such transcribed activates collateral activity Cas12a enzyme, leading to downstream cleavage fluorophore/quencher-labeled reporter thus resulting an increase measured fluorescence signal. We have used MAIGRET six different with high sensitivity (detection limit picomolar range) specificity (no signal presence non-target antibodies). can also be adapted competitive approach antigens. With MAIGRET, significantly expand scope applicability sensing approaches potentially enable measurement any molecular which antibody available.

Language: Английский

Citations

0