Digital CRISPR systems for the next generation of nucleic acid quantification

TrAC Trends in Analytical Chemistry, Год журнала: 2023, Номер 159, С. 116917 - 116917

Опубликована: Янв. 6, 2023

Язык: Английский

---

Microfluidic Biosensor Integrated with Signal Transduction and Enhancement Mechanism for Ultrasensitive Noncompetitive Assay of Multiple Mycotoxins

Analytical Chemistry, Год журнала: 2023, Номер 95(20), С. 7993 - 8001

Опубликована: Май 8, 2023

To achieve high-throughput ultrasensitive detection of mycotoxins in food, a functional DNA-guided transition-state CRISPR/Cas12a microfluidic biosensor (named FTMB) was successfully constructed. The signal transduction strategy FTMB has utilized DNA sequences with specific recognition function and activators to form trigger switches. Meanwhile, the system constructed by adjusting composition ratio crRNA activator high response for low concentrations target mycotoxins. On other hand, enhancement efficiently integrated output quantum dots (QDs) fluorescence effect photonic crystals (PCs). construction universal QDs PC films matching bandgap produced significant factor 45.6. Overall, exhibited wide analytic range (10–5–101 ng·mL–1), limit (fg·mL–1), short period (∼40 min), specificity, good precision (coefficients variation <5%), satisfactory practical sample analysis capacity (the consistency HPLC at 88.76%–109.99%). It would provide new reliable solution rapid multiple small molecules fields clinical diagnosis food safety.

Язык: Английский

---

An autocatalytic CRISPR-Cas amplification effect propelled by the LNA-modified split activators for DNA sensing

Nucleic Acids Research, Год журнала: 2024, Номер 52(7), С. e39 - e39

Опубликована: Март 13, 2024

Abstract CRISPR-Cas systems with dual functions offer precise sequence-based recognition and efficient catalytic cleavage of nucleic acids, making them highly promising in biosensing diagnostic technologies. However, current methods encounter challenges complexity, low turnover efficiency, the necessity for sophisticated probe design. To better integrate Cas proteins, we proposed a novel approach called Autocatalysis Amplification driven by LNA-modified Split Activators (CALSA) detection single-stranded DNA (ssDNA) genomic DNA. By introducing split ssDNA activators site-directed trans-cleavage mediated LNA modifications, an autocatalysis-driven positive feedback loop acids based on LbCas12a system was constructed. Consequently, CALSA enabled one-pot real-time cell-free (cfDNA) from different tumor cell lines. Notably, achieved high sensitivity, single-base specificity, remarkably short reaction times. Due to programmability acid circuits, these results highlighted immense potential as powerful tool cascade signal amplification. Moreover, sensitivity specificity further emphasized value diagnostics, opening avenues future clinical applications.

Язык: Английский

---

Recent Advances in Lateral Flow Assays for Viral Protein Detection with Nanomaterial-Based Optical Sensors

Biosensors, Год журнала: 2024, Номер 14(4), С. 197 - 197

Опубликована: Апрель 17, 2024

Controlling the progression of contagious diseases is crucial for public health management, emphasizing importance early viral infection diagnosis. In response, lateral flow assays (LFAs) have been successfully utilized in point-of-care (POC) testing, emerging as a viable alternative to more traditional diagnostic methods. Recent advancements virus detection primarily leveraged methods such reverse transcription–polymerase chain reaction (RT-PCR), transcription–loop-mediated isothermal amplification (RT-LAMP), and enzyme-linked immunosorbent assay (ELISA). Despite their proven effectiveness, these conventional techniques are often expensive, require specialized expertise, consume significant amount time. contrast, LFAs utilize nanomaterial-based optical sensing technologies, including colorimetric, fluorescence, surface-enhanced Raman scattering (SERS), offering quick, straightforward analyses with minimal training infrastructure requirements detecting proteins biological samples. This review describes composition mechanism recent protein detection, categorizing them into fluorescent, SERS-based techniques. progress, developing simple, stable, highly sensitive, selective LFA system remains formidable challenge. Nevertheless, an advanced promises not only enhance clinical diagnostics but also extend its utility environmental monitoring beyond, demonstrating potential revolutionize both healthcare safety.

Язык: Английский

---

Advances in Biosensor Technologies for Infectious Diseases Detection

TrAC Trends in Analytical Chemistry, Год журнала: 2024, Номер unknown, С. 117979 - 117979

Опубликована: Сен. 1, 2024

Язык: Английский

---

Assessment of the whole genome sequencing of Lactiplantibacillus plantarum 13-3 for elucidation of novel bacteriocin producing gene cluster and confirmation of its potential probiotic functionality and safety applications

CyTA - Journal of Food, Год журнала: 2024, Номер 22(1)

Опубликована: Апрель 5, 2024

Язык: Английский

---

Biosensors for Food Spoilage Detection: A Comprehensive Review of Current Advances

Journal of Food Chemistry and Nanotechnology, Год журнала: 2024, Номер 10

Опубликована: Март 11, 2024

Food spoilage is an enormous problem in the food industry because it can result not only wastage of and financial losses but also lead to health-related problems.Therefore, a timely identification deterioration essential greatly help ensure safety, minimize losses, maintain consumer confidence.In recent years, biosensors have evolved into rapid, sensitive, highly reliable methods for detection indicators.Biosensors are composed unique biological elements transducer that enable quantification markers like volatile organic gases, microorganisms, or enzymes.This review article discusses principles, types, sensing mechanisms used this field, highlighting their advantages limitations.The addresses factors responsible which could be detected by biosensor, such as microbial activity, enzymatic processes, metabolite synthesis.Furthermore, presents most advancements signal transduction techniques, components, platforms biosensors.Overall, objectively evaluates current state terms sensitivity, selectivity, applicability different samples highlights challenges prospects field biosensors, including application nanotechnology, miniaturization, data processing improved prediction.

Язык: Английский

---

Development of a Rapid and Sensitive RPA-CRISPR/Cas12a Assay for Non-invasive Pre-implantation Genetic Testing

Analytica Chimica Acta, Год журнала: 2025, Номер unknown, С. 343687 - 343687

Опубликована: Янв. 1, 2025

Язык: Английский

---

Development of an MSPQC Nucleic Acid Sensor Based on CRISPR/Cas9 for the Detection of Mycobacterium tuberculosis

Analytical Chemistry, Год журнала: 2022, Номер 94(32), С. 11409 - 11415

Опубликована: Авг. 5, 2022

Accurate and rapid detection of nucleic acid plays a vital role in the clinical treatment tuberculosis caused by Mycobacterium (M.TB). However, false-negative false-positive results base mismatches could affect accuracy. Inspired unique property CRISPR/Cas9, we proposed new MSPQC M.TB sensor based on CRISPR/Cas9 system, which can distinguish single-base 10 bases from protospacer adjacent motif (PAM) region. In sensor, single-stranded DNA Au interdigital electrodes was used as capture probe for target an initiator hybridization chain reaction (HCR). HCR to generate long double-stranded (dsDNA), span electrodes. recognition components recognize capture/target dsDNA. When existed, hybridized with form dsDNA, be recognized cut CRISPR/Cas9. Thus, connection between off resulted response. no remained not Therefore, reserved. Moreover, silver staining technology utilized improve sensitivity detection. detected using specific sequence fragments 16S rRNA target. The time down 2.3 h. limit (LOD) 30 CFU/mL.

Язык: Английский

---

Detection of Cancer Marker Flap Endonuclease 1 Using One-Pot Transcription-Powered Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Signal Expansion

Analytical Chemistry, Год журнала: 2022, Номер 94(39), С. 13549 - 13555

Опубликована: Сен. 19, 2022

As a critical functional protein in DNA replication and genome stability, flap endonuclease 1 (FEN1) has been considered promising biomarker druggable target for multiple cancers. We report here transcription-powered clustered regularly interspaced short palindromic repeat (CRISPR)/Cas12a signal expansion platform rapid sensitive detection of FEN1. In this method, the probe cleavage by FEN1 generated free 5′ single-stranded which could hybridize with T7 promoter-bearing template trigger extension. Then, CRISPR guide RNA (crRNA) transcribed from extended activated collateral DNase activity Cas12a, releasing fluorophore quenched to result. The high specificity was validated comparing other repair-relevant proteins. limit (LOD) be as low 0.03 mU, is enough detect biological samples. addition, inhibition assay also successfully achieved platform, proving its potential inhibitor screening. summary, study provides novel biosensor analysis new insights into development CRISPR-based biosensors non-nucleic acid targets.

Язык: Английский

---