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1.
BMC Biotechnol ; 24(1): 30, 2024 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-38720310

RESUMEN

BACKGROUND: Venous thromboembolism (VTE), is a noteworthy complication in individuals with gastric cancer, but the current diagnosis and treatment methods lack accuracy. In this study, we developed a t-PAIC chemiluminescence kit and employed chemiluminescence to detect the tissue plasminogen activator inhibitor complex (t-PAIC), thrombin-antithrombin III complex (TAT), plasmin-α2-plasmin inhibitor complex (PIC) and thrombomodulin (TM), combined with D-dimer and fibrin degradation products (FDP), to investigate their diagnostic potential for venous thrombosis in gastric cancer patients. The study assessed variations in six indicators among gastric cancer patients at different stages. RESULTS: The t-PAIC reagent showed LOD is 1.2 ng/mL and a linear factor R greater than 0.99. The reagents demonstrated accurate results, with all accuracy deviations being within 5%. The intra-batch and inter-batch CVs for the t-PAIC reagent were both within 8%. The correlation coefficient R between this method and Sysmex was 0.979. Gastric cancer patients exhibited elevated levels of TAT, PIC, TM, D-D, FDP compared to the healthy population, while no significant difference was observed in t-PAIC. In the staging of gastric cancer, patients in III-IV stages exhibit higher levels of the six markers compared to those in I-II stages. The ROC curve indicates an enhancement in sensitivity and specificity of the combined diagnosis of four or six indicators. CONCLUSION: Our chemiluminescence assay performs comparably to Sysmex's method and at a reduced cost. The use of multiple markers, including t-PAIC, TM, TAT, PIC, D-D, and FDP, is superior to the use of single markers for diagnosing VTE in patients with malignant tumors. Gastric cancer patients should be screened for the six markers to facilitate proactive prophylaxis, determine the most appropriate treatment timing, ameliorate their prognosis, decrease the occurrence of venous thrombosis and mortality, and extend their survival.


Asunto(s)
Mediciones Luminiscentes , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/diagnóstico , Masculino , Persona de Mediana Edad , Mediciones Luminiscentes/métodos , Femenino , Anciano , Antitrombina III/metabolismo , Antitrombina III/análisis , Trombomodulina/sangre , Productos de Degradación de Fibrina-Fibrinógeno/análisis , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , alfa 2-Antiplasmina/metabolismo , alfa 2-Antiplasmina/análisis , Adulto , Fibrinolisina/metabolismo , Fibrinolisina/análisis , Tromboembolia Venosa/diagnóstico , Tromboembolia Venosa/sangre , Péptido Hidrolasas
2.
Biosens Bioelectron ; 258: 116351, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-38705074

RESUMEN

Multifunctional single-atom catalysts (SACs) have been extensively investigated as outstanding signal amplifiers in bioanalysis field. Herein, a type of Fe single-atom catalysts with Fe-nitrogen coordination sites in nitrogen-doped carbon (Fe-N/C SACs) was synthesized and demonstrated to possess both catalase and peroxidase-like activity. Utilizing Fe-N/C SACs as dual signal amplifier, an efficient bipolar electrode (BPE)-based electrochemiluminescence (ECL) immunoassay was presented for determination of prostate-specific antigen (PSA). The cathode pole of the BPE-ECL platform modified with Fe-N/C SACs is served as the sensing side and luminol at the anode as signal output side. Fe-N/C SACs could catalyze decomposition of H2O2 via their high catalase-like activity and then increase the Faraday current, which can boost the ECL of luminol due to the electroneutrality in a closed BPE system. Meanwhile, in the presence of the target, glucose oxidase (GOx)-Au NPs-Ab2 was introduced through specific immunoreaction, which catalyzes the formation of H2O2. Subsequently, Fe-N/C SACs with peroxidase-like activity catalyze the reaction of H2O2 and 4-chloro-1-naphthol (4-CN) to generate insoluble precipitates, which hinders electron transfer and then inhibits the ECL at the anode. Thus, dual signal amplification of Fe-N/C SACs was achieved by increasing the initial ECL and inhibiting the ECL in the presence of target. The assay exhibits sensitive detection of PSA linearly from 1.0 pg/mL to 100 ng/mL with a detection limit of 0.62 pg/mL. The work demonstrated a new ECL enhancement strategy of SACs via BPE system and expands the application of SACs in bioanalysis field.


Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Electrodos , Peróxido de Hidrógeno , Hierro , Límite de Detección , Mediciones Luminiscentes , Luminol , Antígeno Prostático Específico , Catálisis , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Humanos , Luminol/química , Antígeno Prostático Específico/análisis , Antígeno Prostático Específico/sangre , Hierro/química , Glucosa Oxidasa/química , Inmunoensayo/métodos , Oro/química , Peroxidasa/química , Nanopartículas del Metal/química , Nitrógeno/química , Carbono/química , Naftoles
3.
ACS Chem Biol ; 19(5): 1035-1039, 2024 May 17.
Artículo en Inglés | MEDLINE | ID: mdl-38717306

RESUMEN

Red-shifted bioluminescence is highly desirable for diagnostic and imaging applications. Herein, we report a semisynthetic NanoLuc (sNLuc) based on complementation of a split NLuc (LgBiT) with a synthetic peptide (SmBiT) functionalized with a fluorophore for BRET emission. We observed exceptional BRET ratios with diverse fluorophores, notably in the red (I674/I450 > 14), with a brightness that is sufficient for naked eye detection in blood or through tissues. To exemplify its utility, LgBiT was fused to a miniprotein that binds HER2 (affibody, ZHER2), and the selective detection of HER2+ SK-BR-3 cells over HER2- HeLa cells was demonstrated.


Asunto(s)
Mediciones Luminiscentes , Humanos , Células HeLa , Mediciones Luminiscentes/métodos , Luciferasas/genética , Luciferasas/metabolismo , Receptor ErbB-2/metabolismo , Línea Celular Tumoral , Colorantes Fluorescentes/química
4.
Biosens Bioelectron ; 258: 116356, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-38705073

RESUMEN

In this work, the dual-ligand lanthanide metal-organic framework (MOF)-based electrochemiluminescence (ECL) sensor was constructed for the detection of miRNA-128 in glioblastoma (GBM) diagnosis. The luminescent Eu-MOF (EuBBN) was synthesized with terephthalic acid (BDC) and 2-amino terephthalic acid (BDC-NH2) as dual-ligand. Due to the antenna effect, EuBBN with conjugated-π structure exhibited strong luminescent signal and high quantum efficiency, which can be employed as ECL nanoprobe. Furthermore, the novel plasmonic CuS@Au heterostructure array has been prepared. The localized surface plasmon resonance coupling effect of the CuS@Au heterostructure array can amplify the ECL signal of EuBBN significantly. The EuBBN/CuS@Au heterostructure array-based sensing system has been prepared for the detection of miRNA-128 with a wide linear range from 1 fM to 1 nM and a detection limit of 0.24 fM. Finally, miRNA-128 in the clinic GBM tissue sample has been analysis for the distinguish of tumor grade successfully. The results demonstrated that the dual-ligand MOF/CuS@Au heterostructure array-based ECL sensor can provide important support for the development of GBM diagnosis.


Asunto(s)
Técnicas Biosensibles , Europio , Glioblastoma , Oro , Estructuras Metalorgánicas , MicroARNs , MicroARNs/análisis , Glioblastoma/diagnóstico , Humanos , Estructuras Metalorgánicas/química , Técnicas Biosensibles/métodos , Oro/química , Europio/química , Límite de Detección , Mediciones Luminiscentes/métodos , Ligandos , Técnicas Electroquímicas/métodos , Neoplasias Encefálicas/diagnóstico , Ácidos Ftálicos/química , Nanopartículas del Metal/química , Cobre/química
5.
Bioelectrochemistry ; 158: 108714, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38653106

RESUMEN

G-quadruplex (G4) DNA is present in human telomere oligonucleotide sequences. Oxidative damage to telomeric DNA accelerates telomere shortening, which is strongly associated with aging and cancer. Most of the current analyses on oxidative DNA damage are based on ds-DNA. Here, we developed a electrochemiluminescence (ECL) probe for enhanced recognition of oxidative damage in G4-DNA based on DNA-mediated charge transfer (CT), which could specifically recognize damaged sites depending on the position of 8-oxoguanine (8-oxoG). First, a uniform G4-DNA monolayer interface was fabricated; the G4-DNA mediated CT properties were examined using an iridium(III) complex [Ir(ppy)2(pip)]PF6 stacked with G4-DNA as an indicator. The results showed that G4-DNA with 8-oxoG attenuated DNA CT. The topological effects of oxidative damage at different sites of G4-DNA and their effects on DNA CT were revealed. The sensing platform was also used for the sensitive and quantitative detection of 8-oxoG in G4-DNA, with a detection limit of 28.9 fmol. Overall, these findings present a sensitive platform to study G4-DNA structural and stability changes caused by oxidative damage as well as the specific and quantitative detection of oxidation sites. The different damage sites in the G-quadruplex could provide detailed clues for understanding the function of G4-associated telomere functional enzymes.


Asunto(s)
Daño del ADN , ADN , G-Cuádruplex , Guanina , ADN/química , Guanina/análogos & derivados , Guanina/química , Humanos , Oxidación-Reducción , Estrés Oxidativo , Técnicas Biosensibles/métodos , Mediciones Luminiscentes/métodos , Límite de Detección , Técnicas Electroquímicas/métodos
6.
Talanta ; 274: 125934, 2024 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-38574533

RESUMEN

Nowadays, novel and efficient signal amplification strategy in electrochemiluminescence (ECL) platform is urgently needed to enhance the sensitivity of biosensor. In this work, the dual ECL signal enhancement strategy was constructed by the interactions of Pd nanoparticles attached covalent organic frameworks (Pd NPs@COFs) with tris (bipyridine) ruthenium (RuP) and Exonuclease III (Exo.III) cycle reaction. Within this strategy, the COFs composite was generated from the covalent reaction between 2-nitro-1,4-phenylenediamine (NPD) and trialdehyde phloroglucinol (Tp), and then animated by glutamate (Glu) to attach the Pd NPs. Next, the "signal on" ECL biosensor was constructed by the coordination assembly of thiolation capture DNA (cDNA) onto the Pd NPs@COFs modified electrode. After the aptamer recognition of progesterone (P4) with hairpin DNA 1 (HP1), the Exo. III cycle reaction was initiated with HP2 to generate free DNA, which hybridized with cDNA to form double-stranded DNA (dsDNA). For that, the RuP was embedded into the groove of dsDNA and achieved the ultrasensitive detection of P4 with a lower limit of detection (LOD) down to 0.45 pM, as well as the excellent selectivity and stability. This work expands the COFs-based materials application in ECL signal amplification and valuable DNA cyclic reaction in biochemical testing field.


Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Exodesoxirribonucleasas , Nanopartículas del Metal , Estructuras Metalorgánicas , Paladio , Progesterona , Nanopartículas del Metal/química , Estructuras Metalorgánicas/química , Paladio/química , Progesterona/análisis , Progesterona/química , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Exodesoxirribonucleasas/química , Exodesoxirribonucleasas/metabolismo , Límite de Detección , Mediciones Luminiscentes/métodos , Humanos , ADN/química
7.
Talanta ; 274: 126023, 2024 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-38583328

RESUMEN

Dual-potential ratiometric electrochemiluminescence (ECL) is in favor of resistance to environmental interference. However, two kinds of emitters or coreactants, and a wide scan potential range (>2 V) are mandatory. This work developed a new dual-potential ratiometric ECL sensor for detection of carcinoembryonic antigen (CEA) using single emitter (luminol) and single coreactant (H2O2) with a mild potential range from -0.1 to 0.6 V. Luminol could produce a strong cathodic ECL (Ec) induced by hydroxyl radicals (HO‧) from the reduction of H2O2, and a relatively weak anodic ECL (Ea). After the ferrocene modified CEA aptamer (Apt-Fc) was attached, Fc could promote Ea by catalyzing the oxidation of H2O2, and reduce Ec by consuming HO‧. With the cycling amplification of the exonuclease I, CEA could substantially reduce the amount of Apt-Fc, resulting in the decrease of Ea and the rise of Ec. So, the ratio of Ec to Ea (Ec/Ea) was used as the detection signal, realizing the sensitive determination of CEA from 0.1 pg mL-1 to 10 ng mL-1 with a LOD of 41.85 fg mL-1 (S/N = 3). The developed sensor demonstrated excellent specificity, stability and reproducibility, with satisfactory results in practical detection.


Asunto(s)
Aptámeros de Nucleótidos , Antígeno Carcinoembrionario , Técnicas Electroquímicas , Peróxido de Hidrógeno , Mediciones Luminiscentes , Luminol , Antígeno Carcinoembrionario/análisis , Antígeno Carcinoembrionario/sangre , Técnicas Electroquímicas/métodos , Humanos , Mediciones Luminiscentes/métodos , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Luminol/química , Aptámeros de Nucleótidos/química , Límite de Detección , Técnicas Biosensibles/métodos , Metalocenos/química , Compuestos Ferrosos/química
8.
Talanta ; 274: 126025, 2024 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-38574539

RESUMEN

Exposure to bioaerosol contamination has detrimental effects on human health. Recent advances in ATP bioluminescence provide more opportunities for the quantitative detection of bioaerosols. Since almost all active organisms can produce ATP, the amount of airborne microbes can be easily measured by detecting ATP-driven bioluminescence. The accurate evaluation of microorganisms mainly relies on following the four key steps: sampling and enrichment of airborne microbes, lysis for ATP extraction, enzymatic reaction, and measurement of luminescence intensity. To enhance the effectiveness of ATP bioluminescence, each step requires innovative strategies and continuous improvement. In this review, we summarized the recent advances in the quantitative detection of airborne microbes based on ATP bioluminescence, which focuses on the advanced strategies for improving sampling devices combined with ATP bioluminescence. Meanwhile, the optimized and innovative strategies for the remaining three key steps of the ATP bioluminescence assay are highlighted. The aim is to reawaken the prosperity of ATP bioluminescence and promote its wider utilization for efficient, real-time, and accurate detection of airborne microbes.


Asunto(s)
Adenosina Trifosfato , Microbiología del Aire , Mediciones Luminiscentes , Adenosina Trifosfato/análisis , Mediciones Luminiscentes/métodos , Bacterias/aislamiento & purificación , Humanos , Monitoreo del Ambiente/métodos
9.
Anal Chem ; 96(17): 6652-6658, 2024 Apr 30.
Artículo en Inglés | MEDLINE | ID: mdl-38630909

RESUMEN

A low-triggering potential and a narrow-potential window are anticipated to decrease the electrochemical interference and cross talk of electrochemiluminescence (ECL). Herein, by exploiting the low oxidative potential (0.82 V vs Ag/AgCl) of dihydrolipoic acid-capped sliver nanoclusters (DHLA-AgNCs), a coreactant ECL system of DHLA-AgNCs/hydrazine (N2H4) is proposed to achieve efficient and oxidative-reduction ECL with a low-triggering potential of 0.82 V (vs Ag/AgCl) and a narrow-potential window of 0.22 V. The low-triggering-potential and narrow-potential-window nature of ECL can be primarily preserved upon labeling DHLA-AgNCs to probe DNA and immobilizing DHLA-AgNCs onto the Au surface via sandwiched hybridization, which eventually enables a selective ECL strategy for the gene assay at +0.82 V. This gene assay strategy can sensitively determine the gene of human papillomavirus from 10 to 1000 pM with a low limit of detection of 5 pM (S/N = 3) and would open a way to improve the applied ECL bioassay.


Asunto(s)
Técnicas Electroquímicas , Mediciones Luminiscentes , Nanopartículas del Metal , Plata , Ácido Tióctico/análogos & derivados , Plata/química , Técnicas Electroquímicas/métodos , Nanopartículas del Metal/química , Mediciones Luminiscentes/métodos , Humanos , Ácido Tióctico/química , Técnicas Biosensibles/métodos , ADN Viral/análisis , ADN Viral/genética , Límite de Detección
10.
Anal Bioanal Chem ; 416(13): 3251-3260, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38584178

RESUMEN

Multiplexed lateral flow assays (LFAs) offer efficient on-site testing by simultaneously detecting multiple biomarkers from a single sample, reducing costs. In cancer diagnostics, where biomarkers can lack specificity, multiparameter detection provides more information at the point-of-care. Our research focuses on epithelial ovarian cancer (EOC), where STn-glycosylated forms of CA125 and CA15-3 antigens can better discriminate cancer from benign conditions. We have developed a dual-label LFA that detects both CA125-STn and CA15-3-STn within a single anti-STn antibody test line. This utilizes spectral separation of green (540 nm) and blue (450 nm) emitting erbium (NaYF4:Yb3+, Er3+)- and thulium (NaYF4: Yb3+, Tm3+)-doped upconverting nanoparticle (UCNP) reporters conjugated with antibodies against the protein epitopes in CA125 or CA15-3. This technology allows the simultaneous detection of different antigen variants from a single test line. The developed proof-of-concept dual-label LFA was able to distinguish between the ascites fluid samples from diagnosed ovarian cancer patients (n = 10) and liver cirrhosis ascites fluid samples (n = 3) used as a negative control. The analytical sensitivity of CA125-STn for the dual-label LFA was 1.8 U/ml in buffer and 3.6 U/ml in ascites fluid matrix. Here we demonstrate a novel approach of spectrally separated measurement of STn-glycosylated forms of two different cancer-associated protein biomarkers by using UCNP reporter technology.


Asunto(s)
Antígeno Ca-125 , Proteínas de la Membrana , Mucina-1 , Neoplasias Ováricas , Humanos , Antígeno Ca-125/análisis , Femenino , Neoplasias Ováricas/diagnóstico , Glicosilación , Biomarcadores de Tumor/análisis , Antígenos de Carbohidratos Asociados a Tumores/análisis , Mediciones Luminiscentes/métodos , Carcinoma Epitelial de Ovario/diagnóstico , Inmunoensayo/métodos
11.
Talanta ; 275: 126114, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-38631265

RESUMEN

Electrochemiluminescence (ECL) is one of the most sensitive techniques in the field of diagnostics. However, they typically require luminescent labeling and electrode surface biological modification, which is a time-consuming and laborious process involving multiple steps and may also lead to low reaction efficiency. Fabricating label/modification-free biosensors has become one of the most attractive parts for simplifying the ECL assays. In this work, the ECL luminophores carbon dots (CDs) were encapsulated in DNA hydrogel in situ by a simple rolling circle amplification (RCA) reaction. Upon binding of the target DNA, active Cas12a induces a collateral cleavage of the hydrogel's ssDNA backbone, resulting in a programmable degradation of the hydrogel and the release of CDs. By directly measuring the released CDs ECL, a simple and rapid label/modification-free detection of the target HPV-16 was realized. It is noted that this method allowed for 0.63 pM HPV-16 DNA detection without any amplification step, and it could take only ∼60 min for a fast test of a human serum sample. These results showed that our label/modification-free ECL biosensor has great potential for use in simple, rapid, and sensitive point-of-care (POC) detection.


Asunto(s)
Técnicas Biosensibles , ADN Viral , Técnicas Electroquímicas , Mediciones Luminiscentes , Técnicas Biosensibles/métodos , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos , Humanos , ADN Viral/análisis , ADN Viral/sangre , Proteínas Bacterianas/química , Endodesoxirribonucleasas/química , Carbono/química , Papillomavirus Humano 16/genética , Puntos Cuánticos/química , Hidrogeles/química , Límite de Detección , ADN/química , ADN/análisis , Técnicas de Amplificación de Ácido Nucleico/métodos , Proteínas Asociadas a CRISPR
12.
Talanta ; 275: 126125, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-38663066

RESUMEN

The establishment of rapid target analysis methods for cytokeratin fragment antigen 21-1 (CYFRA 21-1) is urgently needed. [Ir(pbi)2(acac)] (pbi = 2-(4-bromophenyl)-1-hydrogen -benzimidazole, acac = acetylacetonate) as traditional electrochemiluminescence (ECL) luminophores has been confined due to its non-negligible dark toxicity and poor water solubility leading to poor biocompatibility and electrical conductivity as an organic molecule. Hence, to overcome this limitation, [Ir(pbi)2(acac)] can be effectively loaded on the polyvinyl alcohol hydrogel modified Ti3C2Tx MXene surface (Ir@Ti3C2Tx-PVA) as sensing platform which can emit high ECL signals. Then, a quenching strategy was proposed to fabricate an ECL sandwich immunosensor using H2O2 as quencher molecules which can generated by Pd@Au0.85Pd0.15. Especially, the generation of O2 to H2O2 can be achieved through a two-electron (2e-) reaction pathway by Pd@Au0.85Pd0.15, to overcome the restriction that the H2O2 was virtually impossible to label or immobilize on the non-enzyme nanomaterials. The proposed ECL assay achieves a response to CYFRA 21-1 within the range of 0.1 pg/mL-100 ng/mL, with a detection limit of 8.9 fg/mL (S/N = 3). This work provided a feasible tactic to seek superior-performance ECL luminophore and quencher consequently set up a novel means to makeup ultrasensitive ECL biosensor, which extended the utilization potential of Ir(pbi)2(acac) in ECL assays.


Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Oro , Peróxido de Hidrógeno , Queratina-19 , Mediciones Luminiscentes , Paladio , Alcohol Polivinílico , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/análisis , Técnicas Electroquímicas/métodos , Paladio/química , Catálisis , Técnicas Biosensibles/métodos , Alcohol Polivinílico/química , Oro/química , Humanos , Mediciones Luminiscentes/métodos , Queratina-19/análisis , Inmunoensayo/métodos , Antígenos de Neoplasias/análisis , Antígenos de Neoplasias/inmunología , Titanio/química , Límite de Detección , Iridio/química , Nanopartículas del Metal/química
13.
Sci Rep ; 14(1): 9710, 2024 04 27.
Artículo en Inglés | MEDLINE | ID: mdl-38678103

RESUMEN

Among the several animal models of α-synucleinopathies, the well-known viral vector-mediated delivery of wild-type or mutated (A53T) α-synuclein requires new tools to increase the lesion in mice and follow up in vivo expression. To this end, we developed a bioluminescent expression reporter of the human A53T-α-synuclein gene using the NanoLuc system into an AAV2/9, embedded or not in a fibroin solution to stabilise its expression in space and time. We first verified the expression of the fused protein in vitro on transfected cells by bioluminescence and Western blotting. Next, two groups of C57Bl6Jr mice were unilaterally injected with the AAV-NanoLuc-human-A53T-α-synuclein above the substantia nigra combined (or not) with fibroin. We first show that the in vivo cerebral bioluminescence signal was more intense in the presence of fibroin. Using immunohistochemistry, we find that the human-A53T-α-synuclein protein is more restricted to the ipsilateral side with an overall greater magnitude of the lesion when fibroin was added. However, we also detected a bioluminescence signal in peripheral organs in both conditions, confirmed by the presence of viral DNA corresponding to the injected AAV in the liver using qPCR.


Asunto(s)
Dependovirus , Fibroínas , Vectores Genéticos , Mediciones Luminiscentes , Ratones Endogámicos C57BL , alfa-Sinucleína , Animales , alfa-Sinucleína/metabolismo , alfa-Sinucleína/genética , Dependovirus/genética , Humanos , Ratones , Mediciones Luminiscentes/métodos , Vectores Genéticos/genética , Fibroínas/metabolismo , Sistema Nervioso Central/metabolismo , Masculino , Luciferasas/metabolismo , Luciferasas/genética
14.
Anal Chem ; 96(15): 5852-5859, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38556977

RESUMEN

A multicolor electrochemiluminescence (ECL) biosensor based on a closed bipolar electrode (BPE) array was proposed for the rapid and intuitive analysis of three prostate cancer staging indicators. First, [Irpic-OMe], [Ir(ppy)2(acac)], and [Ru(bpy)3]2+ were applied as blue, green, and red ECL emitters, respectively, whose mixed ECL emission colors covered the whole visible region by varying the applied voltages. Afterward, we designed a simple Mg2+-dependent DNAzyme (MNAzyme)-driven tripedal DNA walker (TD walker) to release three output DNAs. Immediately after, three output DNAs were added to the cathodic reservoirs of the BPE for incubation. After that, we found that the emission colors from the anode of the BPE changed as a driving voltage of 8.0 V was applied, mainly due to changes in the interfacial potential and faradaic currents at the two poles of the BPE. Via optimization of the experimental parameters, cutoff values of such three indicators at different clinical stages could be identified instantly with the naked eye, and standard precision swatches with multiple indicators could be prepared. Finally, in order to precisely determine the prostate cancer stage, the multicolor ECL device was used for clinical analysis, and the resulting images were then compared with standard swatches, laying the way for accurate prostate cancer therapy.


Asunto(s)
Técnicas Biosensibles , Neoplasias de la Próstata , Masculino , Humanos , Mediciones Luminiscentes/métodos , Fotometría , Neoplasias de la Próstata/diagnóstico , Antígeno Prostático Específico , ADN , Técnicas Biosensibles/métodos , Electrodos , Técnicas Electroquímicas/métodos
15.
Crit Rev Biomed Eng ; 52(3): 41-62, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38523440

RESUMEN

Microfluidic devices are capable of handling 10-9 L to 10-18 L of fluids by incorporating tiny channels with dimensions of ten to hundreds of micrometers, and they can be fabricated using a wide range of materials including glass, silicon, polymers, paper, and cloth for tailored sensing applications. Microfluidic biosensors integrated with detection methods such as electrochemiluminescence (ECL) can be used for the diagnosis and prognosis of diseases. Coupled with ECL, these tandem devices are capable of sensing biomarkers at nanomolar to picomolar concentrations, reproducibly. Measurement at this low level of concentration makes microfluidic electrochemiluminescence (MF-ECL) devices ideal for biomarker detection in the context of early warning systems for diseases such as myocardial infarction, cancer, and others. However, the technology relies on the nature and inherent characteristics of an efficient luminophore. The luminophore typically undergoes a redox process to generate excited species which emit energy in the form of light upon relaxation to lower energy states. Therefore, in biosensor design the efficiency of the luminophore is critical. This review is focused on the integration of microfluidic devices with biosensors and using electrochemiluminescence as a detection method. We highlight the dual role of carbon quantum dots as a luminophore and co-reactant in electrochemiluminescence analysis, drawing on their unique properties that include large specific surface area, easy functionalization, and unique luminescent properties.


Asunto(s)
Técnicas Biosensibles , Microfluídica , Humanos , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos , Técnicas Biosensibles/métodos , Polímeros
16.
Biosens Bioelectron ; 253: 116170, 2024 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-38442619

RESUMEN

In this paper, a proposal of closed bipolar electrode (BPE) and nanozyme based multi-mode biosensing platform is first presented. As a novel integrated chip, multi-mode-BPE (MMBPE) combines enzyme-linked immunoassay (ELISA), electrochemiluminescence (ECL), ECL imaging and light emitting diode (LED) imaging, enabling highly sensitive triple read-out visible detection of cancer embryonic antigen (CEA). The ECL probe Ab2@Au@Co3O4/CoFe2O4 hollow nanocubes (HNCs) with excellent peroxidase (POD) activity is introduced into the BPE cathode through immune adsorption. The Au@Co3O4/CoFe2O4 HNCs can increase the rate of hydrogen peroxide oxidation of TMB, thus promoting the reaction, and can be used for ELISA detection of CEA at different concentrations. The modification of the BPE sensing interface and reporting interface involved the introduction of the luminescent reagent Ru(bpy)32+ to the BPE anode. The decomposition rate of H2O2 increased under the catalytic action of Au@Co3O4/CoFe2O4 HNCs nanozyme, leading to an accelerated electron transfer rate in the MMBPE system and an enhanced ECL signal from Ru(bpy)32+. The LED imaging technology further provides a convenient and visible approach for CEA imaging in which no additional chemicals are needed. The integration of nanoenzymes as the catalytic core in MMBPE system provides impetus, while the combination of nanozymes with BPE expands the application of nanoenzymes in the field of biological analysis. The integration of intelligent chips with multiple modes of detection shows portable, miniaturized, and integrated excellent properties which meets the requirements of modern detection devices and thus offers a flexible approach for determination of nucleic acids, proteins, and cells.


Asunto(s)
Técnicas Biosensibles , Cobalto , Neoplasias , Óxidos , Humanos , Mediciones Luminiscentes/métodos , Peróxido de Hidrógeno/química , Técnicas Biosensibles/métodos , Electrodos
17.
Anal Chem ; 96(11): 4589-4596, 2024 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-38442212

RESUMEN

Herein, novel europium metal-organic gels (Eu-MOGs) with excellent cathode electrochemiluminescence (ECL) emission are first used to construct biosensors for the ultrasensitive detection of miRNA-222. Impressively, N and O elements of organic ligand 2,2':6,2″-terpyridine 4,4',4″-tricarboxylic acid (H3-tctpy) can perfectly coordinate with Eu3+ to form Eu-MOGs, which not only reduce nonradiative transition caused by the intramolecular free rotation of phenyl rings in other MOGs to enhance the ECL signal with extraordinary ECL efficiency as high as 37.2% (vs the [Ru(bpy)3]2+/S2O82- ECL system) but also reinforce ligand-to-metal charge transfer (LMCT) by the strong affinity between Eu3+ and N and O elements to greatly improve the stability of ECL signals. Besides, an improved nucleic acid cascade amplification reaction is developed to greatly raise the conversion efficiency from target miRNA-222 to a DNAzyme-mediated dual-drive DNA walker as output DNA, which can simultaneously shear the specific recognition sites from two directions. In that way, the proposed biosensor can further enhance the detection sensitivity of miRNA-222 with a linear range of 10 aM-1 nM and a detection limit (LOD) of 8.5 aM, which can also achieve an accurate response in cancer cell lysates of MHCC-97L and HeLa. Additionally, the biosensor can be self-regenerated by the folding/unfolding of related triplets with pH changes to simplify experimental operations and reduce the cost. Hence, this work proposed novel MOGs with stable and intense ECL signals for the construction of a renewable ECL biosensor, supplying a reliable detection method in biomarker analysis and disease diagnosis.


Asunto(s)
Técnicas Biosensibles , ADN Catalítico , MicroARNs , Humanos , Europio , Ligandos , ADN/química , Mediciones Luminiscentes/métodos , MicroARNs/análisis , Técnicas Biosensibles/métodos , Geles , Técnicas Electroquímicas/métodos , Límite de Detección
18.
Diagn Microbiol Infect Dis ; 109(2): 116275, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38537505

RESUMEN

BACKGROUND: Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains a global health threat, necessitating faster and more accessible diagnostic methods. This study investigates critical parameters in the application of a commercial ATP bioluminescence assay for the detection of MTB. METHOD: Our objective was to optimize the ATP bioluminescence protocol using BacTiter-Glo™ for MTB, investigating the impact of varying volumes of MTB suspension and reagent on assay sensitivity, evaluating ATP extraction methods, establishing calibration curves, and elucidating strain-specific responses to antimicrobial agents. RESULTS: ATP extraction methods showed no significant improvement over controls. Calibration curves revealed a linear correlation between relative light units (RLU) and colony-forming units (CFU/mL), establishing low detection limits. Antimicrobial testing demonstrated strain-specific responses aligning with susceptibility and resistance patterns. CONCLUSION: Our findings contribute to refining ATP bioluminescence protocols for enhanced MTB detection and susceptibility testing. Further refinements and validation efforts are warranted, holding promise for more efficient diagnostic platforms in the future.


Asunto(s)
Adenosina Trifosfato , Mediciones Luminiscentes , Mycobacterium tuberculosis , Tuberculosis , Mycobacterium tuberculosis/efectos de los fármacos , Adenosina Trifosfato/análisis , Adenosina Trifosfato/metabolismo , Mediciones Luminiscentes/métodos , Humanos , Tuberculosis/diagnóstico , Tuberculosis/microbiología , Sensibilidad y Especificidad , Pruebas de Sensibilidad Microbiana/métodos , Técnicas Bacteriológicas/métodos
19.
ACS Sens ; 9(4): 1992-1999, 2024 Apr 26.
Artículo en Inglés | MEDLINE | ID: mdl-38536770

RESUMEN

The construction of assays is capable of accurately detecting cytokeratin-19 (CYFRA 21-1), which is critical for the rapid diagnosis of nonsmall cell lung cancer. In this work, a novel electrochemiluminescence (ECL) immunosensor based on the co-reaction promotion of luminol@Au@Ni-Co nanocages (NCs) as ECL probe by Ti3C2Tx MXene@TiO2-MoS2 hybrids as co-reaction accelerator was proposed to detect CYFRA 21-1. Ni-Co NCs, as a derivative of Prussian blue analogs, can be loaded with large quantities of Au NPs, luminol, and CYFRA 21-1 secondary antibodies due to their high specific surface area. To further improve the sensitivity of the developed ECL immunosensor, Ti3C2Tx MXene@TiO2-MoS2 hybrids were prepared by in situ growth of TiO2 nanosheets on highly conductive Ti3C2Tx MXene, and MoS2 was homogeneously grown on Ti3C2Tx MXene@TiO2 surfaces by the hydrothermal method. Ti3C2Tx MXene@TiO2-MoS2 hybrids possess excellent catalytic performance on the electro-redox of H2O2 generating more O2·- and obtaining optimal ECL intensity of the luminol/H2O2 system. Under the appropriate experimental conditions, the quantitative detection range of CYFRA 21-1 was from 0.1 pg mL-1 to 100 ng mL-1, and the limit of detection (LOD) was 0.046 pg mL-1. The present sensor has a lower LOD with a wider linear range, which provides a new analytical assay for the early diagnosis of small-cell-type lung cancer labels.


Asunto(s)
Antígenos de Neoplasias , Técnicas Biosensibles , Disulfuros , Técnicas Electroquímicas , Oro , Queratina-19 , Mediciones Luminiscentes , Luminol , Molibdeno , Titanio , Queratina-19/sangre , Queratina-19/inmunología , Titanio/química , Luminol/química , Molibdeno/química , Oro/química , Antígenos de Neoplasias/inmunología , Técnicas Electroquímicas/métodos , Humanos , Técnicas Biosensibles/métodos , Mediciones Luminiscentes/métodos , Inmunoensayo/métodos , Disulfuros/química , Límite de Detección , Níquel/química , Cobalto/química , Nanopartículas del Metal/química , Anticuerpos Inmovilizados/inmunología , Anticuerpos Inmovilizados/química
20.
ACS Sens ; 9(4): 2176-2182, 2024 Apr 26.
Artículo en Inglés | MEDLINE | ID: mdl-38537645

RESUMEN

Efficient and robust electrochemiluminescence (ECL) emitters are crucial for enhancing the ECL immunosensor sensitivity. This study introduces a novel ECL emitter, CoBIM/Cys, featuring a hierarchical core-shell structure. The core of the structure is created through the swift coordination between the sulfhydryl and carboxyl groups of l-cysteine (l-Cys) and cobalt ions (Co2+), while the shell is constructed by sequentially coordinating benzimidazole (BIM) with Co2+. This design yields a greater specific surface area and a more intricate porous structure compared to CoBIM, markedly enhancing mass transfer and luminophore accessibility. Moreover, the l-Cys and Co2+ core introduces Co-S and Co-O catalytic sites, which improve the catalytic decomposition of H2O2, leading to an increased production of hydroperoxyl radicals (OOH•). This mechanism substantially amplifies the ECL performance. Leveraging the competitive interaction between isoluminol and BIM for OOH• during ECL emission, we developed a ratiometric immunosensor for cardiac troponin I (cTnI) detection. This immunosensor demonstrates a remarkably broad detection range (1 pg mL-1 to 10 ng mL-1), a low detection limit (0.4 pg mL-1), and exceptional reproducibility and specificity.


Asunto(s)
Bencimidazoles , Cisteína , Técnicas Electroquímicas , Mediciones Luminiscentes , Troponina I , Bencimidazoles/química , Cisteína/análisis , Cisteína/química , Mediciones Luminiscentes/métodos , Técnicas Electroquímicas/métodos , Inmunoensayo/métodos , Troponina I/análisis , Troponina I/sangre , Humanos , Límite de Detección , Técnicas Biosensibles/métodos , Cobalto/química , Peróxido de Hidrógeno/química
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