CRISPR-associated Plasmonic Colorimeter Method (Ca-PCM): a real-time RGB detection system for gold nanoparticles-based nucleic acid biosensors DOI Creative Commons

Hernán Alarcón-Iniesta,

G. Arana, María López‐Valls

et al.

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1338, P. 343601 - 343601

Published: Dec. 31, 2024

The detection of genetic sequences represents the gold standard procedure for species discrimination, characterisation tumours, and identification pathogens. development new molecular methods, accessible cost effective, is great relevance. Biosensors based on plasmonic nanoparticles, such as nanoparticles (AuNPs), provide a powerful versatile platform highly sensitive, economic, user-friendly label-free sensing. However, readout techniques typically employed with sensors lack temporal kinetic information, which hampers ability to perform quantitative detection. In this study, novel methodology designated 'CRISPR-associated Plasmonic Colorimeter Method' (Ca-PCM), has been developed. This method combines RNA target recognition by CRISPR LwaCas13a, AuNPs' aggregation, real-time colorimetric Red-Green-Blue (RGB) analysis. system registers AuNP's signatures in using RGB colour sensor 3-channel silicon photodiodes having blue, green red sensitivities. acquired signals are automatically analysed an algorithm designed distinguish between positive negative samples correlate spectral patterns aggregation dose-dependent target. addition, combination Ca-PCM previous isothermal amplification allows efficient real clinical applications. We have shown that analysis continuous measurements promising characterise behaviour nanoparticle-based biosensors achieve simplicity cost-effectiveness approach expand possibilities other plasmonic-based their applicability low-resources environments.

Language: Английский

Visualization Methods for Loop Mediated Isothermal Amplification (LAMP) Assays. DOI
Vinni Thekkudan Novi, Anil Kumar Meher, Abdennour Abbas

et al.

The Analyst, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Recent advances in nucleic acid (NA) detection techniques have significantly enhanced the diagnosis of diseases caused by a range pathogens. These NA-based methods that target specific gene sequences for identification offer high specificity. Despite effectiveness polymerase chain reaction (PCR), its requirement sophisticated laboratory settings and expensive equipment restricts accessibility, particularly resource-limited settings. As an alternative, isothermal amplification are highly sought after due to their rapid, sensitive, ability. Among these, loop mediated (LAMP) stands out simplicity, reliability, cost-effectiveness. LAMP operates without need varied temperature cycles, employing simple heating block maintain constant temperature, thus facilitating onsite rapid testing. In LAMP, step is critical as it shows outcome assay. order make technique user-friendly applicable large scale testing, visual where results can be observed with naked eye. This review focuses on recent developments visualization techniques, including more common fluorescence, turbidity, gel electrophoresis methods, well innovations colorimetric applying novel transduction such nanoparticles digital tools. Additionally, various practical applications discussed.

Language: Английский

Citations

0
ISFET Biosensor with Loop-Mediated Isothermal Amplification for Electronic Rapid Detection of Mycoplasma Pneumoniae DOI Creative Commons
Jie Zou, Jie Hu, Yan Shen

et al.

Sensors, Journal Year: 2025, Volume and Issue: 25(5), P. 1562 - 1562

Published: March 4, 2025

Mycoplasma pneumoniae (MP) is the main culprit of community-acquired pneumonia. Commonly used laboratory testing methods have many shortcomings. Serological diagnosis has low sensitivity, causing false negatives, while a quantitative real-time polymerase chain reaction (qPCR) requires large equipment and professional staff. To make up for these shortcomings, we proposed label-free, low-cost, small-sized ion-sensitive field-effect transistor (ISFET) array based on low-buffered loop-mediated isothermal amplification (LAMP) assay. A complementary metal oxide semiconductor (CMOS)-based ISFET with 512 × sensors was in this system, which responds specifically to H+ sensitivity 365.7 mV/pH. For on-chip amplification, LAMP system designed conserved sequences two genes, CARDS gyrB, applied. The rapid release amounts solution led speedy increase electrical signals captured by array, eliminating need sophisticated temperature cycling optical system. results showed that device can accurately complete MP detection limit about 103 copies/mL (approximately 1 copy per reaction). In final clinical validation, eight throat swab samples using were fully consistent diagnostic outcomes, confirming accuracy reliability use settings. And entire process from sample lysis result interpretation takes 60 min. This platform potential be point-of-care (POCT) pathogen infections, providing basis timely adjustment treatment plans.

Language: Английский

Citations

0
A Route to the Colorimetric Detection of Alpha-Fetoprotein Based on a Smartphone DOI Creative Commons
Junjie Liu,

Qingfubo Geng,

Zhaoxin Geng

et al.

Micromachines, Journal Year: 2024, Volume and Issue: 15(9), P. 1116 - 1116

Published: Aug. 31, 2024

Alpha-fetoprotein (AFP) is a key marker for early cancer detection and assessment. However, the current methods struggle to balance accuracy with need decentralized medical treatment. To address this issue, new AFP analysis platform utilizing digital image colorimetry has been developed. Functionalized gold nanoparticles act as colorimetric agents, changing from purple-red light gray-blue when exposed different concentrations. A smartphone app captures these color changes calculates concentration in sample. improve accuracy, hardware device ensures uniform illumination. Testing confirmed that system can quantitatively analyze using colorimetry. The limit of reached 0.083 ng/mL, average 90.81%. This innovative method enhances testing by offering portability, precision, low cost, making it particularly suitable resource-limited areas.

Language: Английский

Citations

3
An Ultra-Compact and Low-Cost LAMP-Based Virus Detection Device DOI Creative Commons
Dong Guo, Zhengrong Ling, Yifeng Tang

et al.

Sensors, Journal Year: 2024, Volume and Issue: 24(15), P. 4912 - 4912

Published: July 29, 2024

Timely and accurate detection of viruses is crucial for infection diagnosis treatment. However, it remains a challenge to develop portable device that meets the requirement being portable, powerless, user-friendly, reusable, low-cost. This work reports compact ∅30 × 48 mm powerless isothermal amplification (material cost ∼$1 USD) relying on LAMP (Loop-Mediated Isothermal Amplification). We have proposed chromatographic-strip-based microporous permeation technology which can precisely control water flow rate regulate exothermic reaction. heating combined with phase-change materials maintain constant temperature between 50 70 °C duration up 49.8 min. Compared conventional methods, avoids use an additional insulation layer heat preservation, greatly reducing size cost. also deployed color card corresponding algorithm facilitate recognition, data analysis, storage using mobile phone. The experimental results demonstrate our exhibits same limit (LOD) as ProFlex PCR SARS-CoV-2 pseudovirus samples, both 103 copies/μL, verifying its effectiveness reliability. offers timely, low-cost, easy way respiratory infectious disease detection, could provide support in curbing virus transmission protecting health humans animals, especially remote mountainous areas without access electricity or trained professionals.

Language: Английский

Citations

0
Enhancing Pregnancy Care: Harnessing Label‐Free Immunosensors for Pre‐Eclampsia Detection Using PdNPs/Poly(3,5‐Diaminobenzoic Acid) Modified Glassy Carbon Electrodes DOI

Sithara Soman,

Kala Ramakrishnan,

Tripti S Warrier

et al.

ChemistrySelect, Journal Year: 2024, Volume and Issue: 9(37)

Published: Sept. 30, 2024

Abstract Placental Growth factor (PlGF) is one of the biomarkers useful for detecting pregnancy hypertension disorder, Pre‐eclampsia. Herein, we focus on developing a label‐free immunosensor using simple electrochemical methods to detect PlGF antigens pre‐eclampsia diagnosis. The work includes deposition Palladium nanoparticles (PdNPs) glassy carbon electrode (GCE) surface amperometry followed by electropolymerization diaminobenzoic acid (DABA) onto PdNPs/GCE. Antibody (aPlGF) was covalently immobilized pDABA/PdNPs/GCE EDC/NHS reaction. interaction antibody with antigen measured differential pulse voltammetric technique. linear range obtained 1–25 ng mL −1 limit detection 53 pg . stability and reproducibility sensor were in acceptable range. developed used artificial blood urine samples spike recovery analysis.

Language: Английский

Citations

0
CRISPR-associated Plasmonic Colorimeter Method (Ca-PCM): a real-time RGB detection system for gold nanoparticles-based nucleic acid biosensors DOI Creative Commons

Hernán Alarcón-Iniesta,

G. Arana, María López‐Valls

et al.

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1338, P. 343601 - 343601

Published: Dec. 31, 2024

The detection of genetic sequences represents the gold standard procedure for species discrimination, characterisation tumours, and identification pathogens. development new molecular methods, accessible cost effective, is great relevance. Biosensors based on plasmonic nanoparticles, such as nanoparticles (AuNPs), provide a powerful versatile platform highly sensitive, economic, user-friendly label-free sensing. However, readout techniques typically employed with sensors lack temporal kinetic information, which hampers ability to perform quantitative detection. In this study, novel methodology designated 'CRISPR-associated Plasmonic Colorimeter Method' (Ca-PCM), has been developed. This method combines RNA target recognition by CRISPR LwaCas13a, AuNPs' aggregation, real-time colorimetric Red-Green-Blue (RGB) analysis. system registers AuNP's signatures in using RGB colour sensor 3-channel silicon photodiodes having blue, green red sensitivities. acquired signals are automatically analysed an algorithm designed distinguish between positive negative samples correlate spectral patterns aggregation dose-dependent target. addition, combination Ca-PCM previous isothermal amplification allows efficient real clinical applications. We have shown that analysis continuous measurements promising characterise behaviour nanoparticle-based biosensors achieve simplicity cost-effectiveness approach expand possibilities other plasmonic-based their applicability low-resources environments.

Language: Английский

Citations

0