RESUMEN
Loofah sponge-like carbon nanofibers (LF-Co,N/CNFs) were utilized as a carrier for Ru(bpy)32+, and then combined with CdS to create a novel solid-state electrochemiluminescence sensor capable of detecting trace amounts of fenpropathrin. LF-Co,N/CNFs, obtained through the high-temperature pyrolysis of ZIF-67 coaxial electrospinning fibers, were characterized by a loofah-like morphology and exhibited a significant specific surface area and porosity. Apart from serving as a carrier, LF-Co,N/CNFs also functioned as a luminescence accelerator, enhancing the system's luminescence efficiency by facilitating electron transmission and reducing the transmission distance. The inclusion of CdS in the luminescence reaction, in conjunction with Ru(bpy)32+, further boosted the sensor's luminescence signal. The resulting sensor demonstrated a satisfactory signal, with fenpropathrin causing significant quenching of the ECL signal. Under optimized conditions, a linear relationship between the signal quench value and fenpropathrin concentration in the range 1 × 10-12 to 1 × 10-6 M was observed, with a detection limit of 3.3 × 10-13 M (S/N = 3). This developed sensor is characterized by its simplicity, sensitivity, and successful application in detecting fenpropathrin in real samples. The study not only presents a straightforward detection platform for fenpropathrin but also introduces new avenues for the rapid determination of various food contaminants, thereby expanding the utility of carbon fibers in electrochemiluminescence sensors.
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Carbono , Técnicas Electroquímicas , Límite de Detección , Mediciones Luminiscentes , Nanofibras , Nanofibras/química , Mediciones Luminiscentes/métodos , Carbono/química , Técnicas Electroquímicas/métodos , Técnicas Electroquímicas/instrumentación , Animales , Contaminación de Alimentos/análisis , Compuestos de Cadmio/química , Piretrinas/análisis , Compuestos OrganometálicosRESUMEN
The oriented design of reticular materials as emitters can significantly enhance the sensitivity of electrochemiluminescence (ECL) sensing analysis for disease markers. However, due to the structural fragility of hydrogen bonds, relational research on hydrogen-bonded organic frameworks (HOFs) has not been thoroughly conducted. Additionally, the modulation of luminescence behavior through HOFs has been rarely reported. In view of this, hydrogen-bonded biohybrid organic frameworks (HBOFs) were synthesized and recruited for ECL immunoassay applications. HBOFs was easily prepared using 6,6',6â³,6â´-(pyrene-1,3,6,8-tetrayl)tetrakis(2-naphthoic acid) as linkers via bovine serum albumin (BSA) activated hydrogen-bonded cross-linking. The material exhibited good fluorescence emission characteristics. And the highly ordered topological structure and molecular motion limitation mediated by BSA overcome aggregation-caused quenching and generate strong aggregation induced emission, expressing hydrogen-bond interaction enhanced ECL (HIE-ECL) activity with the participation of tri-n-propylamine. Furthermore, a sandwich immunosensor was constructed employing cobalt-based metal-phenolic network (CMPN) coated ferrocene nanoparticles (FNPs) as quenchers (CMPN@FNPs). Signal closure can be achieved by annihilating the excited state through electron transfer from both CMPN and FNPs. Using a universal disease marker, carcinoembryonic antigen, as the analysis model, the signal-off sensor obtained a detection limit of 0.47 pg/mL within the detection range of 1 pg/mL - 50 ng/mL. The synthesis and application of highly stable HBOFs triggered by proteins provide a reference for the development of new reticular ECL signal labels, and electron transfer model provides flexible solutions for more sensitive sensing analysis.
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Técnicas Biosensibles , Técnicas Electroquímicas , Enlace de Hidrógeno , Mediciones Luminiscentes , Albúmina Sérica Bovina , Técnicas Biosensibles/métodos , Inmunoensayo/métodos , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Humanos , Albúmina Sérica Bovina/química , Animales , Estructuras Metalorgánicas/química , Límite de Detección , Bovinos , Metalocenos/química , Compuestos Ferrosos/química , Anticuerpos Inmovilizados/química , Biomarcadores/análisis , Cobalto/químicaRESUMEN
BACKGROUND: Chemiluminescence (CL) bioassay is one of the most advanced and used detection method in clinical diagnosis and biomedical research because of the advantages of low background, easy operation, and wide-field imaging without a light source or microscope. The luminol/hydrogen peroxide/horseradish peroxidase (luminol/H2O2/HRP) system is the most popular CL system, but its application in high-throughput imaging detection is challenged due to its low luminescence efficiency and flash-type emission which is difficult in ensuring the reproducibility and consistency of detection results. RESULTS: We reported a glow-type CL system of luminol@CD/H2O2/HRP by using a supramolecular enhancer of cyclodextrin (CD). This luminol@CD/H2O2/HRP system exhibited a luminescence lifetime of 41 min for sensitive and accurate imaging analysis. The long-lasting CL emission was attributed to the formation of a 1:1 host-guest complex between luminol and CD, which could stabilize the emitter and effectively reduce nonradiative relaxation. The formation of luminol@CD complex was determined through NMR experiments and theoretical analysis. Under optimum conditions, the luminol@CD/H2O2/HRP system showed higher sensitivity and much better precision than classical luminol/H2O2/HRP system for imaging detection of HRP. Especially, this glow-type luminol@CD/H2O2/HRP system realized CL imaging of microwell arrays on microfluidic chips. In addition, the luminol@CD/H2O2/HRP system was successfully applied for point-of-care detection of 17ß-estradiol based on a competitive mechanism of host-guest recognition. SIGNIFICANCE: An efficient CL system is crucial for obtaining reproducible and consistent results for accurate detection. Our luminol@CD/H2O2/HRP system emitted strong and persistent luminescence, resulting in reliability and efficiency at both CL macroscopic and microscopic imaging detection. We expected the luminol@CD/H2O2/HRP CL system to be applied in various detection fields.
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Ciclodextrinas , Peroxidasa de Rábano Silvestre , Peróxido de Hidrógeno , Mediciones Luminiscentes , Luminol , Luminol/química , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Ciclodextrinas/química , Mediciones Luminiscentes/métodos , Peroxidasa de Rábano Silvestre/química , Peroxidasa de Rábano Silvestre/metabolismo , Humanos , Luminiscencia , Límite de DetecciónRESUMEN
A Fe2+-EGTA(ethylene glycol-bis (ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid)-H2O2 system emits photons, and quenching this chemiluminescence can be used for determination of anti-hydroxyl radical (â¢OH) activity of various compounds. The generation of â¢OH and light emission due to oxidative damage to EGTA may depend on the buffer and pH of the reaction milieu. In this study, we evaluated the effect of pH from 6.0 to 7.4 (that may occur in human cells) stabilized with 10 mM phosphate buffer (main intracellular buffer) on a chemiluminescence signal and the ratio of this signal to noise (light emission from medium alone). The highest signal (4698 ± 583 RLU) and signal-to-noise ratio (9.7 ± 1.5) were noted for pH 6.6. Lower and higher pH caused suppression of these variables to 2696 ± 292 RLU, 4.0 ± 0.8 at pH 6.2 and to 3946 ± 558 RLU, 5.0 ± 1.5 at pH 7.4, respectively. The following processes may explain these observations: enhancement and inhibition of â¢OH production in lower and higher pH; formation of insoluble Fe(OH)3 at neutral and alkaline environments; augmentation of â¢OH production by phosphates at weakly acidic and neutral environments; and decreased regeneration of Fe2+-EGTA in an acidic environment. Fe2+-EGTA-H2O2 system in 10 mM phosphate buffer pH 6.6 seems optimal for the determination of anti-â¢OH activity.
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Ácido Egtácico , Peróxido de Hidrógeno , Peróxido de Hidrógeno/química , Concentración de Iones de Hidrógeno , Humanos , Ácido Egtácico/química , Ácido Egtácico/análogos & derivados , Radical Hidroxilo/química , Hierro/química , Luminiscencia , Mediciones Luminiscentes/métodos , LuzRESUMEN
A smartphone-assisted electrochemiluminescence (ECL) strategy based on Ru(bpy)2(L)4+ as chromophores confined with metal - organic frameworks (Ru(bpy)2(L)4+@MOF-5) for the signal-amplified detection of miRNA-21 was developed. We synthesized a derivative of tris(2,2'-bipyridyl)ruthenium(II) complex (Ru(bpy)2(L)4+) with high charges, which can be loaded into the MOF-5 by strong electrostatic interaction to prevent from leakage. In addition, nucleic acid cycle amplification was used to quench the signal of Ru(bpy)2(L)4+@MOF-5 by ferrocene. This method was applied to detect the concentration of miRNA-21 ranging from 1.0 × 10-14-1.0 × 10-9 M with a low LOD of 7.2 fM. This work demonstrated the construction of a signal quenching strategy ECL biosensor for miRNA using Ru(bpy)2(L)4+@MOF-5 systems and its application in smartphone-assisted ECL detection.
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Técnicas Biosensibles , Técnicas Electroquímicas , Límite de Detección , Mediciones Luminiscentes , Estructuras Metalorgánicas , MicroARNs , Teléfono Inteligente , MicroARNs/análisis , MicroARNs/sangre , Técnicas Biosensibles/métodos , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos , Técnicas Electroquímicas/instrumentación , Estructuras Metalorgánicas/química , Humanos , Complejos de Coordinación/química , Compuestos Organometálicos/químicaRESUMEN
BACKGROUND: Persistent infection with human papillomavirus (HPV) significantly contributes to the development of cervical cancer. Thus, it is urgent to develop rapid and accurate methods for HPV detection. Herein, we present an ultrasensitive CRISPR/Cas12a-based electrochemiluminescent (ECL) imaging technique for the detection of HPV-18 DNA. RESULT: The ECL DNA sensor array is constructed by applying black hole quencher (BHQ) and polymer dots (Pdots) co-labeled hairpin DNA (hpDNA) onto a gold-coated indium tin oxide slide (Au-ITO). The ECL imaging method involves an incubation process of target HPV-18 with a mixture of crRNA and Cas12a to activate Cas12a, followed by an incubation of the active Cas12a with the ECL sensor. This interaction causes the indiscriminate cleavage of BHQ from Pdots by digesting hpDNA on the sensor surface, leading to the restoration of the ECL signal of Pdots. The ECL brightness readout demonstrates superior performance of the ECL imaging technique, with a linear detection range of 10 fM-500 pM and a limit-of-detection (LOD) of 5.3 fM. SIGNIFICANCE: The Cas12a-based ECL imaging approach offers high sensitivity and a broad detection range, making it highly promising for nucleic acid detection applications.
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Sistemas CRISPR-Cas , Técnicas Electroquímicas , Mediciones Luminiscentes , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Sistemas CRISPR-Cas/genética , Humanos , Técnicas Biosensibles/métodos , ADN Viral/análisis , ADN Viral/genética , Papillomavirus Humano 18/genética , Límite de Detección , Oro/química , Proteínas Asociadas a CRISPR , Proteínas Bacterianas , EndodesoxirribonucleasasRESUMEN
Bioluminescence imaging (BLI) is an important noninvasive optical imaging technique that has been widely used to monitor many biological processes due to its high sensitivity, resolution, and signal-to-noise ratio. However, the BLI technique based on the firefly luciferin-luciferase system is limited by the expression of exogenous luciferase and the short half-life of firefly luciferin, which pose challenges for long-term tracking in vivo. To solve the problems, here we rationally designed an intelligent strategy for persistent BLI in tumors by combining luciferase-loaded calcium phosphate nanoparticles (Luc@CaP NPs) to provide luciferase and the probe Cys(SEt)-Lys-CBT (CKCBT) to slowly produce the luciferase substrate amino luciferin (Am-luciferin). Luc@CaP NPs constructed with CaP as a carrier could enable luciferase activity to be maintained in vivo for at least 12 h. And compared to the conventional substrate luciferin, CKCBT apparently prolonged the BL time by up to 2 h through GSH-induced intracellular self-assembly and subsequent protease degradation-induced release of Am-luciferin. We anticipate that this strategy could be applied for clinical translation in more disease diagnosis and treatment in the near future.
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Neoplasias de la Mama , Fosfatos de Calcio , Luciferasas , Mediciones Luminiscentes , Nanopartículas , Fosfatos de Calcio/química , Nanopartículas/química , Animales , Mediciones Luminiscentes/métodos , Humanos , Luciferasas/metabolismo , Luciferasas/química , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/patología , Femenino , Ratones , Imagen Óptica , Ratones Endogámicos BALB C , Línea Celular Tumoral , Ratones DesnudosRESUMEN
Mouse orthotopic xenograft tumor models are commonly employed in studies investigating the mechanisms underlying the development and progression of tumors and their preclinical treatment. However, the unavailability of mature and visualized orthotopic xenograft models of nasopharyngeal carcinoma limits the development of treatment strategies for this cancer. The aim of this study was to provide a simple and reliable method for building an orthotopic xenograft model of nasopharyngeal carcinoma. Human nasopharyngeal carcinoma (C666-1-luc) cells, stably expressing the firefly luciferase gene, were injected subcutaneously into the right axilla of BALB/C nude mice. Four weeks later, the resulting subcutaneous tumors were cut into small blocks and grafted into the nasopharynx of immunodeficient BALB/C nude mice to induce tumor formation. Tumor growth was monitored by bioluminescence imaging and small animal magnetic resonance imaging (MRI). The expression of histological and immunological antigens associated with orthotopic xenograft nasopharyngeal carcinoma was analyzed by tissue section analysis and immunohistochemistry (IHC). A visualized orthotopic xenograft nasopharyngeal carcinoma model was successfully developed in this study. Luminescence signal detection, micro-MRI, and hematoxylin and eosin staining revealed the successful growth of tumors in the nasopharynx of the nude mice. Moreover, IHC analysis detected cytokeratin (CK), CK5/6, P40, and P63 expression in the orthotopic tumors, consistent with the reported expression of these antigens in human nasopharyngeal tumors. This study established a reproducible, visual, and less lethal orthotopic xenograft model of nasopharyngeal carcinoma, providing a platform for preclinical research.
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Modelos Animales de Enfermedad , Ratones Desnudos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas , Animales , Humanos , Carcinoma Nasofaríngeo/patología , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/diagnóstico por imagen , Ratones , Neoplasias Nasofaríngeas/patología , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/diagnóstico por imagen , Neoplasias Nasofaríngeas/genética , Línea Celular Tumoral , Ratones Endogámicos BALB C , Carcinoma/patología , Carcinoma/genética , Carcinoma/metabolismo , Imagen por Resonancia Magnética/métodos , Ensayos Antitumor por Modelo de Xenoinjerto , Xenoinjertos , Mediciones Luminiscentes/métodosRESUMEN
BACKGROUND: Plasma p-tau217 has emerged as the most promising blood-based marker (BBM) for the detection of Alzheimer Disease (AD) pathology, yet few studies have evaluated plasma p-tau217 performance in memory clinic settings. We examined the performance of plasma p-tau217 for the detection of AD using a high-sensitivity immunoassay in individuals undergoing diagnostic lumbar puncture (LP). METHODS: Paired plasma and cerebrospinal fluid (CSF) samples were analysed from the TIMC-BRAiN cohort. Amyloid (Aß) and Tau (T) pathology were classified based on established cut-offs for CSF Aß42 and CSF p-tau181 respectively. High-sensitivity electrochemiluminescence (ECL) immunoassays were performed on paired plasma/CSF samples for p-tau217, p-tau181, Glial Fibrillary Acidic Protein (GFAP), Neurofilament Light (NfL) and total tau (t-tau). Biomarker performance was evaluated using Receiver-Operating Curve (ROC) and Area-Under-the-Curve (AUC) analysis. RESULTS: Of 108 participants (age: 69 ± 6.5 years; 54.6% female) with paired samples obtained at time of LP, 64.8% (n = 70/108) had Aß pathology detected (35 with Mild Cognitive Impairment and 35 with mild dementia). Plasma p-tau217 was over three-fold higher in Aß + (12.4 pg/mL; 7.3-19.2 pg/mL) vs. Aß- participants (3.7 pg/mL; 2.8-4.1 pg/mL; Mann-Whitney U = 230, p < 0.001). Plasma p-tau217 exhibited excellent performance for the detection of Aß pathology (AUC: 0.91; 95% Confidence Interval [95% CI]: 0.86-0.97)-greater than for T pathology (AUC: 0.83; 95% CI: 0.75-0.90; z = 1.75, p = 0.04). Plasma p-tau217 outperformed plasma p-tau181 for the detection of Aß pathology (z = 3.24, p < 0.001). Of the other BBMs, only plasma GFAP significantly differed by Aß status which significantly correlated with plasma p-tau217 in Aß + (but not in Aß-) individuals. Application of a two-point threshold at 95% and 97.5% sensitivities & specificities may have enabled avoidance of LP in 58-68% of cases. CONCLUSIONS: Plasma p-tau217 measured using a high-sensitivity ECL immunoassay demonstrated excellent performance for detection of Aß pathology in a real-world memory clinic cohort. Moving forward, clinical use of plasma p-tau217 to detect AD pathology may substantially reduce need for confirmatory diagnostic testing for AD pathology with diagnostic LP in specialist memory services.
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Enfermedad de Alzheimer , Péptidos beta-Amiloides , Biomarcadores , Proteínas tau , Humanos , Proteínas tau/sangre , Proteínas tau/líquido cefalorraquídeo , Femenino , Péptidos beta-Amiloides/sangre , Péptidos beta-Amiloides/líquido cefalorraquídeo , Masculino , Anciano , Biomarcadores/sangre , Biomarcadores/líquido cefalorraquídeo , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/líquido cefalorraquídeo , Inmunoensayo/métodos , Persona de Mediana Edad , Estudios de Cohortes , Mediciones Luminiscentes/métodosRESUMEN
The sensitivity and specificity of electrochemiluminescence (ECL)-based biosensor directly rely on the property of luminophor, the type of sensing carriers and the effectiveness of signal amplification used in the sensor design, which poses a major challenge to manage these elements simultaneously. In this work, an aggregation-induced electrochemiluminescence (AIECL) microfluidic sensing chip using 4',4â³,4â´,4â´'-(ethene-1,1,2,2-tetrayl)tetrabiphenyl-4-carboxylic acid (TPE)-derived hafnium-based metal-organic framework (Hf-MOF) as emitter was developed. An easily overlooked marine pollutant, okadaic acid (OA) with different concentrations ranging from 5.00 ng/mL to 1.50 × 104 ng/mL at the electrode is visualized imaging benefit from high luminescence efficiency of Hf-MOF coupled the rolling circle amplification strategy assisted by trans-cleavage activity of CRISPR/Cas12a. These highlights will solve the long-lasting task in the accurate analysis of small molecule pollutants, which can be able to provide more worthy reference solution about construction of novel ECL luminophor and signal extraction of low-abundance disease-related biomarkers.
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Técnicas Biosensibles , Técnicas Electroquímicas , Mediciones Luminiscentes , Estructuras Metalorgánicas , Ácido Ocadaico , Técnicas Biosensibles/métodos , Técnicas Biosensibles/instrumentación , Mediciones Luminiscentes/métodos , Mediciones Luminiscentes/instrumentación , Técnicas Electroquímicas/métodos , Ácido Ocadaico/análisis , Estructuras Metalorgánicas/química , Dispositivos Laboratorio en un Chip , Límite de Detección , Diseño de EquipoRESUMEN
Multimodality reporter gene imaging combines the sensitivity, resolution and translational potential of two or more signals. The approach has not been widely adopted by the animal imaging community, mainly because its utility in this area is unproven. We developed a new complementation-based reporter gene system where the large component of split NanoLuc luciferase (LgBiT) presented on the surface of cells (TM-LgBiT) interacts with a radiotracer consisting of the high-affinity complementary HiBiT peptide labeled with a radionuclide. Radiotracer uptake could be imaged in mice using SPECT/CT and bioluminescence within two hours of implanting reporter-gene-expressing cells. Imaging data were validated by ex vivo biodistribution studies. Following the demonstration of complementation between the TM-LgBiT protein and HiBiT radiotracer, we validated the use of the technology in the highly specific in vivo multimodal imaging of cells. These findings highlight the potential of this new approach to facilitate the advancement of cell and gene therapies from bench to clinic.
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Genes Reporteros , Luciferasas , Animales , Ratones , Luciferasas/metabolismo , Luciferasas/genética , Humanos , Distribución Tisular , Imagen Óptica/métodos , Mediciones Luminiscentes/métodos , Tomografía Computarizada por Tomografía Computarizada de Emisión de Fotón Único/métodos , Cintigrafía/métodos , Línea Celular TumoralRESUMEN
Primary aldosteronism (PA) is the most common cause of endocrine arterial hypertension, and the suggested screening test for case detection is the aldosterone-to-renin ratio (ARR) or aldosterone-to-direct renin ratio (ADRR) based on radio-immunoassay (RIA) and chemiluminescence assay (CLIA), respectively. The objective of our study was to evaluate the reliability of CLIA for aldosterone and renin measurement and the diagnostic performance of ADRR. A prospective cohort of 1110 patients referred to a single laboratory medicine center underwent measurement of aldosterone and direct renin concentration (DRC) by CLIA and measurement of aldosterone and plasma renin activity (PRA) by RIA. Of 1110 patients, 640 obtained a final diagnosis of hypertension, and 90 of these patients were diagnosed with PA. Overall, between-method correlation was highly significant for aldosterone concentrations (R = 0.945, p < 0.001) and less strong but significant for DRC/PRA (R = 0.422, p < 0.001). Among hypertensive patients, in PA cases, the areas under the receiver operator characteristics (ROC) curves were 0.928 (95% confidence interval 0.904-0.954) for ADRR and 0.943 (95% confidence interval 0.920-0.966) for ARR and were comparable and not significantly different. The highest accuracy was obtained with an ADRR cut-off of 25 (ng/L)/(mIU/L), displaying a sensitivity of 91% and a specificity of 85%. The chemiluminescence assay for aldosterone and DRC is a reliable method for PA diagnosis compared to the classical RIA method.
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Aldosterona , Hiperaldosteronismo , Mediciones Luminiscentes , Renina , Humanos , Hiperaldosteronismo/diagnóstico , Hiperaldosteronismo/sangre , Aldosterona/sangre , Renina/sangre , Femenino , Persona de Mediana Edad , Masculino , Mediciones Luminiscentes/métodos , Adulto , Curva ROC , Estudios Prospectivos , Hipertensión/sangre , Hipertensión/diagnóstico , Anciano , Reproducibilidad de los ResultadosRESUMEN
A new chemiluminescence immunoassay method (CLIA) for detecting IgA anti-transglutaminase (atTG IgA) in celiac disease (CD) has prompted inquiries into its diagnostic performance. We conducted a systematic review and meta-analysis comparing CLIA with traditional enzyme-linked immunosorbent assay (ELISA) and fluorescence enzyme immunoassay (FEIA). We searched PubMed, Medline, and Embase databases up to March 2024. The diagnostic references were intestinal biopsy and ESPGHAN guidelines. We calculated the sensitivity and specificity of atTG IgA assessed by CLIA and the odds ratio (OR) between the assays. Eleven articles were eligible for the systematic review and seven for the meta-analysis. Sensitivity and specificity of atTG IgA CLIA-assay were 0.98 (95% CI, 0.95-0.99) and 0.97 (95% CI, 0.94-0.99), respectively. The sensitivity of atTG IgA antibody detection did not significantly vary across the three assay modalities examined (CLIA vs. ELISA OR: 1.08 (95% CI, 0.56-2.11; p = 0.8); CLIA vs. FEIA OR: 6.97 (95% CI, 0.60-81.03; p = 0.1). The specificity of atTG IgA assessed by FEIA was higher than for CLIA (OR 0.17 (95% CI, 0.05-0.62); p < 0.007). According to the systematic review, normalization of atTG IgA levels in CD patients following a gluten-free diet was delayed when using CLIA compared to ELISA and FEIA methods. Conflicting findings were reported on the antibody threshold to use in order to avoid biopsy confirmation.
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Enfermedad Celíaca , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina A , Mediciones Luminiscentes , Transglutaminasas , Humanos , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/inmunología , Transglutaminasas/inmunología , Inmunoglobulina A/sangre , Mediciones Luminiscentes/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Sensibilidad y Especificidad , Autoanticuerpos/sangre , LuminiscenciaRESUMEN
High excitation potential is recognized as a harmful factor for the biological activity of biomacromolecules, such as proteins and nucleic acids, in electrochemiluminescence (ECL) biosensing. Developing low-potential ECL luminophores is vital for improving ECL accuracy in actual sample sensing. In this work, based on porous metal-organic framework (MOF) structure with multiple active sites and energy transfer between the excited ligands and Ln nodes, we designed a series of Ln-MOFs and observed ECL emission at low potential, providing a novel method to realize low-potential ECL. The MOF nanoemitters were prepared using 1,3,5-tri (4-carboxyphenyl)benzene ligand and several lanthanide ions as nodes through mild hydrothermal reaction. Interestingly, strong ECL emission at +0.75 V of peak potential was observed in the ECL-potential curve of Tb-based MOF using 2,2',2â³-nitrilotriethanol as coreactant, which was beneficial for reducing background interference in biosensing, and this ECL emission was attributed to the energy transfer between Tb and excited ligand. This low-potential ECL was then applied to construct an ECL biosensor with newly developed Cas12a-based method for selective detection of microRNA-155 without the help of strand displacement or reverse transcription. For this ECL system, the limit of detection was 0.78 nM, and the overall detection time was 2.5 h. The Ln-MOF nanoemitter provides a robust ECL platform to selectively detect various targets by integrating new bio-related techniques.
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Técnicas Electroquímicas , Mediciones Luminiscentes , Estructuras Metalorgánicas , MicroARNs , Terbio , Humanos , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Electrodos , Límite de Detección , Mediciones Luminiscentes/métodos , Estructuras Metalorgánicas/química , MicroARNs/análisis , Terbio/químicaRESUMEN
In this work, the copper nanoclusters (Cu NCs) were confined on 3D layered double hydroxide (3D-LDH) to form Cu NCs@3D-LDH with outstanding electrochemiluminescence (ECL) for constructing ultrasensitive biosensor to detect of glial fibrillary acidic protein (GFAP) implicated in Alzheimer's Disease (AD). More importantly, compared to the individual Cu NCs, Cu NCs@3D-LDH presented strong and stable ECL response, since 3D-LDH could not only gather more Cu NCs but also limit the intramolecular free motion to reduce nonradiative transition for obtaining high ECL intensity. In addition, the improved cascade amplification method combining proximity ligation assay (PLA) with DNAzyme could transform tiny amount of target protein into a large amount of output DNA to improve sensitivity of biosensor. The ECL biosensor realized ultrasensitive detection of GFAP with the detection limit of 2 ag/mL and it had been successfully applied to the evaluation of GFAP in the serum of patients with neurological diseases. This research offered a general and facile method to improve ECL performance of Cu NCs for sensitive detection of biomarkers for disease diagnosis.
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Técnicas Biosensibles , Cobre , Técnicas Electroquímicas , Proteína Ácida Fibrilar de la Glía , Hidróxidos , Límite de Detección , Mediciones Luminiscentes , Nanopartículas del Metal , Cobre/química , Técnicas Biosensibles/métodos , Humanos , Proteína Ácida Fibrilar de la Glía/sangre , Proteína Ácida Fibrilar de la Glía/análisis , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Hidróxidos/química , Nanopartículas del Metal/química , Enfermedad de Alzheimer/sangre , Enfermedad de Alzheimer/diagnósticoRESUMEN
Electrochemiluminescence (ECL) sensing systems have surged in popularity in recent years, making significant strides in sensing and biosensing applications. The realization of high-throughput ECL sensors hinges on the implementation of novel signal amplification strategies, propelling the field toward a new era of ultrasensitive analysis. A key strategy for developing advanced ECL sensors and biosensors involves utilizing novel structures with remarkable properties. The past few years have witnessed the emergence of MXenes as a captivating class of 2D materials, with their unique properties leading to exploitation in diverse applications. This review provides a comprehensive summary of the latest advancements in MXene-modified materials specifically engineered for ECL sensing and biosensing applications. We thoroughly analyze the structure, surface functionalization, and intrinsic properties of MXenes that render them exceptionally suitable candidates for the development of highly sensitive ECL sensors and biosensors. Furthermore, this study explores the broad spectrum of applications of MXenes in ECL sensing, detailing their multifaceted roles in enhancing the performance and sensitivity of ECL (bio)sensors. By providing a comprehensive overview, this review is expected to promote progress in related areas.
Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Mediciones Luminiscentes , Nanocompuestos , Técnicas Biosensibles/métodos , Técnicas Biosensibles/instrumentación , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Nanocompuestos/química , HumanosRESUMEN
Serious adverse drug reactions of gentamicin (GM) significantly limit its clinical use, thus there is an urgent demand to develop reliable strategies to detect its concentration. In this study, we have developed a novel highly sensitive and portable lateral flow immunoassay (LFIA) based on CoFe PBAs/WS2 nanozyme mediated chemiluminescence (CL) and photothermal (PT) dual-mode POCT biosensor for the detection of GM, which successfully combines sensitive laboratory analyses with portable in situ analyses in the field. In this proof-of-principle work, the dynamic detection ranges of CL-LFIA and PT-LFIA mode were 1 pg mL-1 to 100 ng mL-1 and 50 pg mL-1 to 100 ng mL-1 with the limits of detection of 0.33 and 16.67 pg mL-1, respectively. The whole detection of CL-LFIA and PT-LFIA could be completed within 15 min and 30 min, respectively. The recoveries of GM spiked into complex matrices including milk, urine, and serum for CL-LFIA and PT-LFIA were 90.94%-109.74% and 94.49%-109.31%, respectively, indicating the reliability and applicability of the dual-mode LFIA in real samples. The dual-mode POCT biosensor could effectively overcome the false problems with improving accuracy and sensitivity, enabling user to precisely detect GM by laboratory analysis or on-site analysis depending on the source condition. Due to the complementary properties of CL-LFIA and PT-LFIA, the developed POCT biosensor can effectively ensure high-performance detection, showing the potential application of accurately detecting drug concentration in clinical practice.
Asunto(s)
Técnicas Biosensibles , Gentamicinas , Límite de Detección , Mediciones Luminiscentes , Pruebas en el Punto de Atención , Gentamicinas/análisis , Gentamicinas/sangre , Técnicas Biosensibles/métodos , Inmunoensayo/métodos , Mediciones Luminiscentes/métodos , Humanos , Animales , Antibacterianos/análisis , Antibacterianos/sangre , Antibacterianos/orina , Leche/química , Cobalto/químicaRESUMEN
Patulin (PAT) is a widespread fruit toxin. Trace-level PAT exposure can cause serious harm to human health. Herein, a multimodal PAT aptasensor was designed based on Ru(bpy)32+-based metal organic framework composited hydrogel (RuMOF@hydrogel) and versatile banana peel-derived carbonized polymer dots (BPPDs). RuMOF@hydrogel modified magnetic-electrode exhibited excellent anodic and cathodic electrochemiluminescence (ECL) emission and stability. Meanwhile, the BPPDs could enhance anodic ECL of RuMOF@hydrogel, and also show excellent fluorescence (FL) and photothermal (PT) properties. With the aid of PAT-triggered hybridization chain reaction and magnetic separation, ECL, FL, and PT responses could be recorded concurrently. The detection limit can reach as low as 0.25 fg mL-1. The ratiometric ECL quantitation ensured the sensitivity and accuracy of this assay. And visual FL and portable PT modes contributed to the utility. Furthermore, this aptasensor demonstrated better performances than HPLC in fruit products and the protocol can be extended to determine various contaminants in foods.
Asunto(s)
Técnicas Biosensibles , Contaminación de Alimentos , Frutas , Patulina , Polímeros , Frutas/química , Patulina/análisis , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Contaminación de Alimentos/análisis , Polímeros/química , Hidrogeles/química , Aptámeros de Nucleótidos/química , Límite de Detección , Estructuras Metalorgánicas/química , Técnicas Electroquímicas/instrumentación , Técnicas Electroquímicas/métodos , Puntos Cuánticos/química , Musa/química , Mediciones Luminiscentes/instrumentación , Mediciones Luminiscentes/métodosRESUMEN
There is a pressing need for new antibiotics to combat rising resistance to those already in use. The bacterial general secretion (Sec) system has long been considered a good target for novel antimicrobials thanks to its irreplacable role in maintaining cell envelope integrity, yet the lack of a robust, high-throughput method to screen for Sec inhibition has so far hampered efforts to realize this potential. Here, we have adapted our recently developed in vitro assay for Sec activityâbased on the split NanoLuc luciferaseâto work at scale and in living cells. A simple counterscreen allows compounds that specifically target Sec to be distinguished from those with other effects on cellular function. As proof of principle, we have applied this assay to a library of 5000 compounds and identified a handful of moderately effective in vivo inhibitors of Sec. Although these hits are unlikely to be potent enough to use as a basis for drug development, they demonstrate the efficacy of the screen. We therefore anticipate that the methods presented here will be scalable to larger compound libraries, in the ultimate quest for Sec inhibitors with clinically relevant properties.
Asunto(s)
Antibacterianos , Ensayos Analíticos de Alto Rendimiento , Antibacterianos/farmacología , Antibacterianos/química , Ensayos Analíticos de Alto Rendimiento/métodos , Escherichia coli/efectos de los fármacos , Escherichia coli/metabolismo , Luminiscencia , Mediciones Luminiscentes/métodos , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/metabolismo , Canales de Translocación SEC/antagonistas & inhibidores , Canales de Translocación SEC/metabolismoRESUMEN
Ganoderma lucidum (G. lucidum) is a traditional edible fungus with strong medicinal value. G. lucidum polysaccharides (GLP) encapsulate many of the key beneficial properties of this species, providing a valuable tool for the treatment of a range of diseases. The present study was developed to explore the protective benefits of GLP treatment in the context of arsenic poisoning. Through microscopy and flow cytometry experiments, NaAsO2 was found to induce rat tracheal epithelial (RTE) cell apoptosis, together with reductions in cell surface epidermal growth factor receptor (EGFR) expression. GLP treatment, however, was able to reduce apoptosis rates and elevate the expression of EGFR relative to NaAsO2-treated cells. GLP extracts (50, 100, 200 mg·mL-1) prepared from four types of G. lucidum were administered to RTE cells damaged with arsenic, revealing limited differences in position resistance among these varieties. This work provides reference for the pharmaceutical and medical research of G. lucidum.