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1.
Adv Mater ; 36(15): e2312540, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38288781

RESUMO

On-site diagnostic tests that accurately identify disease biomarkers lay the foundation for self-healthcare applications. However, these tests routinely rely on single-mode signals and suffer from insufficient accuracy, especially for multiplexed point-of-care tests (POCTs) within a few minutes. Here, this work develops a dual-mode multiclassification diagnostic platform that integrates an electrochemiluminescence sensor and a field-effect transistor sensor in a microfluidic chip. The microfluidic channel guides the testing samples to flow across electro-optical sensor units, which produce dual-mode readouts by detecting infectious biomarkers of tuberculosis (TB), human rhinovirus (HRV), and group B streptococcus (GBS). Then, machine-learning classifiers generate three-dimensional (3D) hyperplanes to diagnose different diseases. Dual-mode readouts derived from distinct mechanisms enhance the anti-interference ability physically, and machine-learning-aided diagnosis in high-dimensional space reduces the occasional inaccuracy mathematically. Clinical validation studies with 501 unprocessed samples indicate that the platform has an accuracy approaching 99%, higher than the 77%-93% accuracy of rapid point-of-care testing technologies at 100% statistical power (>150 clinical tests). Moreover, the diagnosis time is 5 min without a trade-off of accuracy. This work solves the occasional inaccuracy issue of rapid on-site diagnosis, endowing POCT systems with the same accuracy as laboratory tests and holding unique prospects for complicated scenes of personalized healthcare.


Assuntos
Sistemas Automatizados de Assistência Junto ao Leito , Testes Imediatos , Humanos , Microfluídica , Biomarcadores
2.
Adv Sci (Weinh) ; 11(6): e2307840, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38070186

RESUMO

Early diagnosis of acute diseases is restricted by the sensitivity and complex process of sample treatment. Here, an ultrasensitive, rapid, and portable electrochemiluminescence-microfluidic (ECL-M) system is described via sandwich-type immunoassay and surface plasmonic resonance (SPR) assay. Using a sandwich immunoreaction approach, the ECL-M system employs cardiac troponin-I antigen (cTnI) as a detection model with a Ru@SiO2 NPs labeled antibody as the signal probe. For miR-499-5p detection, gold nanoparticles generate SPR effects to enhance Ru(bpy)3 2+ ECL signals. The system based on alternating current (AC) electroosmotic flow achieves an LOD of 2 fg mL-1 for cTnI in 5 min and 10 aM for miRNAs in 10 min at room temperature. The point-of-care testing (POCT) device demonstrated 100% sensitivity and 98% specificity for cTnI detection in 123 clinical serum samples. For miR-499-5p, it exhibited 100% sensitivity and 97% specificity in 55 clinical serum samples. Continuous monitoring of these biomarkers in rats' saliva, urine, and interstitial fluid samples for 48 hours revealed observations rarely documented in biotic fluids. The ECL-M POCT device stands as a top-performing system for ECL analysis, offering immense potential for ultrasensitive, rapid, highly accurate, and facile detection and monitoring of acute diseases in POC settings.


Assuntos
Nanopartículas Metálicas , MicroRNAs , Ratos , Animais , Eletro-Osmose , Ouro , Dióxido de Silício , Doença Aguda , Microfluídica , Técnicas Eletroquímicas , Medições Luminescentes
3.
Anal Chem ; 95(42): 15755-15762, 2023 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-37824574

RESUMO

In this study, we developed an innovative highly specific nucleic acid isothermal detection assay based on prokaryotic DNA polymerase I with exquisitely designed fluorescent probes, achieving high sensitivity and 100% specificity within 30 min. The fluorescent nucleic acid probe was designed and constructed based on the specific flap cleavage endonuclease activity of prokaryotic DNA polymerase I (including the Bst, Bsu, Bsm, and Klenow DNA polymerases). The flap endonuclease activity depends on the length of the flap DNA and polymerization activity, which greatly reduces the false-positive rate caused by primer dimerization. This robust assay was also validated by the detection of rotavirus with great specificity and sensitivity. It could be a great alternative to qPCR in the field of point-of-care detection of pathogens.


Assuntos
DNA Polimerase I , Técnicas de Amplificação de Ácido Nucleico , Polimerização , DNA , Endonucleases , Sensibilidade e Especificidade
4.
Biosensors (Basel) ; 13(8)2023 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-37622869

RESUMO

Considering the high risk of heavy metal ions (HMIs) transferring through the food chain and accumulating in milk, a flexible and facile point-of-care testing (POCT) platform is urgently needed for the accurate, sensitive, and highly selective on-site quantification of multiple HMIs in milk. In this work, a cost-effective disk with six screen-printed electrodes (SPEs) was designed for hand-held electrochemical detection. Metal organic frameworks (MOFs) were adopted to amplify and enhance the electrochemical signals of methylene blue (MB). Using differential pulse voltammetry (DPV) methods, low limits of detection for four HMIs (Cd2+, 0.039 ppb; Hg2+, 0.039 ppb; Pb2+, 0.073 ppb; and As3+, 0.022 ppb) were achieved within four minutes. Moreover, the quantitative POCT system was applied to milk samples. The advantages of low cost, ease of on-site implementation, fast response, and accuracy allow for the POCT platform to be used in practical monitoring applications for the quantitation of multiple HMIs in milk samples.


Assuntos
Estruturas Metalorgânicas , Metais Pesados , Animais , Azul de Metileno , Leite , Testes Imediatos , Íons
5.
Chem Commun (Camb) ; 57(92): 12270-12272, 2021 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-34751285

RESUMO

An automated Cas12a-microfluidic system was constructed to distinguish the B.1.617.2 (delta) variant of SARS-CoV-2 from the wild-type virus rapidly and was validated using 30 clinical samples, showing 100% consistency with next-generation sequencing. It will be a potential tool for the rapid differential diagnosis of the delta variant of SARS-CoV-2.


Assuntos
COVID-19/diagnóstico , Sistemas CRISPR-Cas/genética , Microfluídica/métodos , SARS-CoV-2/genética , Automação , COVID-19/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação , Reação em Cadeia da Polimerase , RNA Guia de Cinetoplastídeos/genética , RNA Guia de Cinetoplastídeos/metabolismo , RNA Viral/metabolismo , SARS-CoV-2/isolamento & purificação
6.
ACS Appl Mater Interfaces ; 12(7): 7974-7983, 2020 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-31985197

RESUMO

We report a general fabrication method for water-soluble pillar[n]arene nanosponges (NS) by the use of a supra-amphiphilic template. For the first time, a supra-amphiphilic template is used to conveniently control the size of host molecule-based NS. The intrinsic cavity of water-soluble pillar[6]arene could stably encapsulate dyes (acridine orange and indocyanine green) and antitumor drugs (doxorubicin hydrochloride and mitoxantrone) by host-guest interaction. NS could deliver antitumor drugs to cancer cells. Multidrug resistance (MDR) of cancer cells (MCF-7/ADR) is overcome by the use of NS with a ninefold reduction in the IC50 value compared to that of the free drug (3.4 µM vs 34.4 µM). Mechanistic studies show that stable encapsulation of the antitumor drug is the reason to overcome MDR.


Assuntos
Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Mitoxantrona/farmacologia , Nanoestruturas/química , Compostos de Amônio Quaternário/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Resistência a Múltiplos Medicamentos , Humanos , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Microscopia Eletrônica de Transmissão , Nanoestruturas/toxicidade , Nanoestruturas/ultraestrutura , Tamanho da Partícula , Água/química
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