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Here, a multiplex surface-enhanced Raman scattering (SERS)-immunochromatography (ICA) platform is presented using a graphene oxide (GO)-based film-like magnetic tag (GFe-DAu-D/M) that effectively captures and detects multiple bacteria in complex specimens. The 2D GFe-DAu-D/M tag with universal bacterial capture ability is fabricated through the layer-by-layer assembly of one layer of small Fe3O4 nanoparticles (NPs) and two layers of 30 nm AuNPs with a 0.5 nm built-in nanogap on monolayer GO nanosheets followed by co-modification with 4-mercaptophenylboronic acid (MPBA) and 5,5'-dithiobis-(2-nitrobenzoic acid).The GFe-DAu-D/M enabled the rapid enrichment of multiple bacteria by MPBA and quantitative analysis of target bacteria on test lines by specific antibodies, thus achieving multiple signal amplification of magnetic enrichment effect and multilayer dense hotspots and eliminating matrix interference in real-world applications. The developed technology can directly and simultaneously diagnose three major pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella typhimurium) with detection limits down to the level of 10 cells mL-1. The good performance of the proposed method in the detection of real urinary tract infection specimens is also demonstrated, suggesting the great potential of the GFe-DAu-D/M-ICA platform for the highly sensitive monitoring of bacterial infections or contamination.
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Bactérias , Grafite , Análise Espectral Raman , Análise Espectral Raman/métodos , Grafite/química , Bactérias/isolamento & purificação , Cromatografia de Afinidade/métodos , Ouro/química , Humanos , Nanopartículas de Magnetita/química , Staphylococcus aureus/isolamento & purificaçãoRESUMO
In this paper, the aluminium-doped carbon dots (Al-CDs) were developed for simultaneous selective detection of five tetracycline antibiotics (TCs), including minocycline (MC), tetracycline (TC), oxytetracycline (OTC), doxycycline (DOC) and chlortetracycline (CTC). With the bright blue fluorescence, Al-CDs displayed excellent stability and showed no obvious fluorescence intensity changes under different ionic strength, acidic or alkaline environment, continuous ultraviolet light illumination, and even longtime storage at room temperature. As adding different antibiotics, the fluorescence of Al-CDs was strongly quenched by five TCs and showed no distinguished changes with the addition of other kinds of antibiotics. The presence of interferential metal ions, anions and small organic molecules imposed no effect on the simultaneous selective detection of five TCs. A good linear relationship was achieved for five TCs in the range of 0-100 µM, and the limit of detection for MC, TC, OTC, DOC, and CTC were 13.91 (0-100 µM), 15.54 (0-100 µM), 14.26 (0-100 µM), 13.48 (0-100 µM) and 13.88 nM (0-100 µM), respectively. Moreover, Al-CDs was successfully used to the detection of five TCs in real samples with recovery ranging from 92.47% to 122.05%, confirming a bright future for the practical applications in the assays of foods, medicines, and environments.
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Tuning fork, consisting of two fork arms and a fork handle, has a stable and rigid structure. Inspired by this structure, a tuning fork-shaped DNA (TF-DNA) fluorescence aptasensor was constructed to detect ochratoxin A (OTA) and aflatoxin B1 (AFB1). A TF-DNA double-stranded structure capable of attaching both OTA aptamer labeled with the FAM fluorescent group (FAM-Apt) and AFB1 aptamer labeled with the ROX fluorescent group (ROX-Apt) was designed and linked to magnetic beads. This TF-DNA double-stranded structure can provide a stable platform for dual-target detection. In the presence of OTA and AFB1, FAM-Apt and ROX-Apt preferentially bound to them and detached from the TF-DNA double-stranded structure. Dual-signal fluorescent probes were collected from the supernatant by magnetic separation, and achieved fluorescence enhancement at 520 nm and 607 nm, respectively. The linear ranges are 0.05 ng/mL to 100 ng/mL for OTA and 0.1 ng/mL to 100 ng/mL for AFB1, and the detection limits are 0.015 ng/mL and 0.045 ng/mL, respectively. The developed sensor has the advantages of simple and fast preparation, good specificity and reproducibility, which is promising for the simultaneous determination of multiple hazardous substances in food.
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The developed method for simultaneous detection of aflatoxin B1 (AFB1) and aflD genes can effectively monitor from the source and reduce the safety problems and economic losses caused by the production of aflatoxin, which can be of great significance for food safety regulations. In this paper, we constructed a sensitive and convenient fluorescent biosensor to detect AFB1 and aflD genes simultaneously based on fluorescence resonance energy transfer (FRET) between quantum dots (QDs) and a black hole quenching agent. A stable "Y" shaped aptasensor was employed as the detection platform and a double quantum dot labeled DNA fragment was utilized to be the sensing element in this work. When the targets of AFB1 and aflD genes were presented in the solution, the aptamer in the "Y" shaped probe is specifically recognized by the target. At this time, both Si-carbon quantum dots (Si-CDs) and CdTe QDs are far away from the BHQ1 and BHQ3 to recover the fluorescence. The linear range of the prepared fluorescence simultaneous detection method was as wide as 0.5-500 ng·mL-1 with detection lines of 0.64 ng·mL-1 for AFB1 and 0.5-500 nM with detection lines of 0.75 nM for aflD genes (3σ/k). This fabricated fluorescent biosensor was further validated in real rice flour and corn flour samples, which also achieved good results. The recoveries were calculated by comparing the known and found amounts of AFB1 which ranged from 88.4 to approximately 115.32% in the rice flour samples and 90.7 ~ 102.58% in the corn flour samples. The recoveries of aflD genes ranged from 84.32 to approximately 109.3% in the rice flour samples and 89.48 ~ 100.99% in the corn flour samples. Therefore, the proposed biosensor can significantly improve food safety and quality control through a simple, fast, and sensitive agricultural product monitoring and detection system.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Compostos de Cádmio , Pontos Quânticos , Aflatoxina B1/análise , Aptâmeros de Nucleotídeos/genética , Telúrio , Corantes Fluorescentes , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
The application of antimony sulfide sensors, characterized by their exceptional stability and selectivity, is of emerging interest in detection research, and the integration of graphitized carbon materials is expected to further enhance their electrochemical performance. This study represents a pioneering effort in the synthesis of carbon-doped antimony sulfide materials through the pyrolysis of the mixture of microorganisms and their synthetic antimony sulfide. The prepared materials are subsequently applied to electrochemical sensors for monitoring the highly toxic compounds catechol (CC) and hydroquinone (HQ) in the environment. Via cyclic voltammetry (CV) and impedance testing, we concluded that the pyrolytic product at 700 °C (Sb-700) demonstrated the best electrochemical properties. Differential pulse voltammetry (DPV) revealed impressive separation when utilizing Sb-700/GCE for simultaneous detection of CC and HQ, exhibiting good linearity within the concentration range of 0.1-140 µM. The achieved sensitivities of 24.62 µA µM-1 cm-2 and 22.10 µA µM-1 cm-2 surpassed those of most CC and HQ electrochemical sensors. Meanwhile, the detection limits for CC and HQ were as low as 0.18 µM and 0.16 µM (S/N = 3), respectively. Additional tests confirmed the good selectivity, reproducibility, and long-term stability of Sb-700/GCE, which was effective in detecting CC and HQ in tap water and river water, with recovery rates of 100.7%-104.5% and 96.5%-101.4%, respectively. It provides a method that combines green microbial synthesis and simple pyrolysis for the preparation of electrode materials in CC and HQ electrochemical sensors, and also offers a new perspective for the application of microbial synthesized materials.
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Antimônio , Catecóis , Técnicas Eletroquímicas , Hidroquinonas , Pirólise , Hidroquinonas/química , Hidroquinonas/análise , Catecóis/análise , Catecóis/química , Antimônio/química , Antimônio/análise , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Sulfetos/químicaRESUMO
BACKGROUND: Feline herpesvirus type 1 (FHV) and Feline calicivirus (FCV) are the primary co-infecting pathogens that cause upper respiratory tract disease in cats. However, there are currently no visual detection assays available for on-site testing. Here, we develop an ultrasensitive and visual detection method based on dual recombinase polymerase amplification (dRPA) reaction and the hybrid Cas12a/Cas13a trans-cleavage activities in a one-tube reaction system, referred to as one-tube dRPA-Cas12a/Cas13a assay. RESULTS: The recombinant plasmid DNAs, crRNAs, and RPA oligonucleotides targeting the FCV ORF1 gene and FHV-1 TK gene were meticulously prepared. Subsequently, dual RPA reactions were performed followed by screening of essential reaction components for hybrid CRISPR-Cas12a (targeting the FHV-1 TK gene) and CRISPR-Cas13a (targeting the FCV ORF1 gene) trans-cleavage reaction. As a result, we successfully established an ultra-sensitive and visually detectable method for simultaneous detection of FCV and FHV-1 nucleic acids using dRPA and CRISPR/Cas-powered technology in one-tube reaction system. Visual readouts were displayed using either a fluorescence detector (Fluor-based assay) or lateral flow dipsticks (LDF-based assay). As expected, this optimized assay exhibited high specificity towards only FHV-1 and FCV without cross-reactivity with other feline pathogens while achieving accurate detection for both targets with limit of detection at 2.4 × 10- 1 copies/µL for the FHV-1 TK gene and 5.5 copies/µL for the FCV ORF1 gene, respectively. Furthermore, field detection was conducted using the dRPA-Cas12a/Cas13a assay and the reference real-time PCR methods for 56 clinical samples collected from cats with URTD. Comparatively, the results of Fluor-based assay were in exceptional concordance with the reference real-time PCR methods, resulting in high sensitivity (100% for both FHV-1 and FCV), specificity (100% for both FHV-1 and FCV), as well as consistency (Kappa values were 1.00 for FHV-1 and FCV). However, several discordant results for FHV-1 detection were observed by LDF-based assay, which suggests its prudent use and interpretaion for clinical detection. In spite of this, incorporating dRPA-Cas12a/Cas13a assay and visual readouts will facilitate rapid and accurate detection of FHV-1 and FCV in resource-limited settings. CONCLUSIONS: The one-tube dRPA-Cas12a/Cas13a assay enables simultaneously ultrasensitive and visual detection of FHV-1 and FCV with user-friendly modality, providing unparalleled convenience for FHV-1 and FCV co-infection surveillance and decision-making of URTD management.
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Calicivirus Felino , Herpesviridae , Varicellovirus , Gatos , Animais , Recombinases/genética , Sistemas CRISPR-CasRESUMO
The Strep Easy Kit, a bio-enrichment dual ICG-strip test, is a diagnostic tool designed for the detection of Streptococcus agalactiae, an important pathogenic bacterium in tilapia farming. The kit can simultaneously identify two different serotypes of S. agalactiae, serotype Ia and serotype III. This capability is crucial for the collection of valuable epidemiological data and facilitates strategic planning for effective vaccine development and deployment. The Strep Easy Kit consists of two main steps: pathogen enrichment and pathogen detection. The enrichment step increases the concentration of bacteria so that the bacterial load is raised to the level reliably detectable by the subsequent ICG strip test. This is achieved by incubating the fish samples in a suitable liquid medium under specified temperature and time conditions. The second step involves the use of the dual-ICG strip test. This strip test consists of two monoclonal antibodies and one polyclonal antibody that are specific to S. agalactiae and can distinguish between S. agalactiae serotype Ia and S. agalactiae serotype III. This dual capability enables the ICG strip test to simultaneously detect both serotypes of S. agalactiae in a single test kit. The detection limit of the test kit, which consists of a dual ICG-Strip test combined with an enrichment step, is 100 CFU/mL. The kit can be used to detect S. agalactiae in both live and dead fish samples, making it versatile for various testing scenarios. The test results obtained using the Strep Easy Kit have shown a 94.4% correlation with the standard method (Thai Agricultural Standard; TAS 10453-2010), with 90.2% sensitivity and 100% specificity. Significant advantages of the Strep Easy Kit lie in its simplicity and portability, allowing farmers to perform the test by themselves and on-site. This makes it a practical and accessible tool for the tilapia farming industry.
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A dual-template molecularly imprinted electrochemical sensor was developed for the simultaneous detection of serotonin (5-HT) and glutamate (Glu). First, amino-functionalized reduced graphene oxide (NRGO) was used as the modification material of a GCE to increase its electrical conductivity and specific surface area, using Glu and 5-HT as dual-template molecules and o-phenylenediamine (OPD) with self-polymerization ability as functional monomers. Through self-assembly and electropolymerization, dual-template molecularly imprinted polymers were formed on the electrode. After removing the templates, the specific recognition binding sites were exposed. The amount of NRGO, polymerization parameters, and elution parameters were further optimized to construct a dual-template molecularly imprinted electrochemical sensor, which can specifically recognize double-target molecules Glu and 5-HT. The differential pulse voltammetry (DPV) technique was used to achieve simultaneous detection of Glu and 5-HT based on their distinct electrochemical activities under specific conditions. The sensor showed a good linear relationship for Glu and 5-HT in the range 1 ~ 100 µM, and the detection limits were 0.067 µM and 0.047 µM (S/N = 3), respectively. The sensor has good reproducibility, repeatability, and selectivity. It was successfully utilized to simultaneously detect Glu and 5-HT in mouse serum, offering a more dependable foundation for objectively diagnosing and early warning of depression. Additionally, the double signal sensing strategy also provides a new approach for the simultaneous detection of both electroactive and non-electroactive substances.
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Técnicas Eletroquímicas , Ácido Glutâmico , Grafite , Limite de Detecção , Impressão Molecular , Fenilenodiaminas , Serotonina , Serotonina/sangue , Serotonina/análise , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Animais , Ácido Glutâmico/análise , Ácido Glutâmico/sangue , Ácido Glutâmico/química , Grafite/química , Camundongos , Fenilenodiaminas/química , Depressão/diagnóstico , Depressão/sangue , Eletrodos , Biomarcadores/sangue , Biomarcadores/análise , Reprodutibilidade dos TestesRESUMO
Nitrofuran (NF) contamination in food products is a global problem resulting in the banned utilization and importation of nitrofuran contaminated products. A novel chromogenic detection method using a specific DNA aptamer with high affinity and specificity to nitrofurans was developed. Single-stranded DNA aptamers specific to nitrofuran metabolites, including 3-amino-2-oxazolidinone (AOZ), 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), and 1-aminohydantoin (AHD), were isolated using magnetic bead-SELEX. The colorimetric detection of nitrofurans using gold nanoparticles (AuNPs) exhibited an AOZ detection range of 0.01-0.06 ppb with a limit of detection (LOD) of 0.03 ppb. At the same time, this system could detect AMOZ and AHD at a range of 0.06 ppb and 10 ppb, respectively. The fast nitrofuran extraction method was optimized for food, such as fish tissues and honey, adjusted to be completed within 3-6 h. This novel apta-chromogenic detection method could detect NF metabolites with a sensitivity below the minimum required performance limit (MPRL). This analysis will be valuable for screening, with a shortened time of detection for aquaculture products such as shrimp and fish muscle tissues.
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Aptâmeros de Nucleotídeos , Contaminação de Alimentos , Nanopartículas Metálicas , Nitrofuranos , Nitrofuranos/análise , Nitrofuranos/metabolismo , Nanopartículas Metálicas/química , Contaminação de Alimentos/análise , Aptâmeros de Nucleotídeos/química , Oxazolidinonas/análise , Oxazolidinonas/metabolismo , Ouro/química , Limite de Detecção , Hidantoínas/análise , Animais , Mel/análise , Colorimetria/métodos , Análise de Alimentos/métodosRESUMO
Dopamine (DA) and uric acid (UA) are essential for many physiological processes in the human body. Abnormal levels of DA and UA can lead to multiple diseases, such as Parkinson's disease and gout. In this work, a three-dimensional reduced graphene oxide-MXene (3D rGO-Ti3C2) composite electrode was prepared using a simple one-step hydrothermal reduction process, which could separate the oxidation potentials of DA and UA, enabling the simultaneous detection of DA and UA. The 3D rGO-Ti3C2 electrode exhibited excellent electrocatalytic activity towards both DA and UA. In 0.01 M PBS solution, the linear range of DA was 0.5-500 µM with a sensitivity of 0.74 µA·µM-1·cm-2 and a detection limit of 0.056 µM (S/N = 3), while the linear range of UA was 0.5-60 µM and 80-450 µM, with sensitivity of 2.96 and 0.81 µA·µM-1·cm-2, respectively, and a detection limit of 0.086 µM (S/N = 3). In 10% fetal bovine serum (FBS) solution, the linear range of DA was 0.5-500 µM with a sensitivity of 0.41 µA·µM-1·cm-2 and a detection limit of 0.091 µM (S/N = 3). The linear range of UA was 2-500 µM with a sensitivity of 0.11 µA·µM-1·cm-2 and a detection limit of 0.6 µM (S/N = 3). The modified electrode exhibited advantages such as high sensitivity, a strong anti-interference capability, and good repeatability. Furthermore, the modified electrode was successfully used for DA measurement in vivo. This could present a simple reliable route for neurotransmitter detection in neuroscience.
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Dopamina , Técnicas Eletroquímicas , Eletrodos , Grafite , Ácido Úrico , Grafite/química , Ácido Úrico/análise , Ácido Úrico/sangue , Dopamina/análise , Dopamina/sangue , Técnicas Eletroquímicas/métodos , Limite de Detecção , Oxirredução , Humanos , Titânio/química , AnimaisRESUMO
Atherosclerosis is a primary global health concern due to its high morbidity and mortality. This disease is characterized by a complex interplay of chronic inflammation, oxidative stress, and proteolytic enzymes. Traditional imaging techniques struggle to capture the dynamic biochemical processes within atherosclerotic plaques. Herein, we have developed a novel unimolecular photoacoustic probe (UMAPP) that combines specific recognition sites for neutrophil elastase (NE) and the redox pair O2â¢â/GSH into a cohesive molecular platform, allowing in vivo monitoring of oxidative stress and activated neutrophils within plaques. UMAPP features a boron-dipyrromethene (BODIPY) core linked to a hydrophilic NE-cleavable tetrapeptide, and dual oxidative stress-responsive catechol moieties, enabling NE-mediated modulation of photoinduced electron transfer, affecting the photoacoustic intensity at 685 nm (PA685), while oxidation and reduction of the catechol groups by O2â¢â and GSH lead to reversible, ratiometric changes in the photoacoustic spectrum. Preliminary applications of UMAPP have successfully differentiated between atherosclerotic and healthy mice, assessed the impact of pneumonia on plaque composition, and validated the probe's efficacy in drug-treatment studies, detecting molecular changes prior to observable histopathological alterations. UMAPP's integrated molecular imaging approach holds significant promise for advancing the diagnosis and management of atherosclerosis by enabling earlier and more precise detection of vulnerable plaques.
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Rapid and accurate imaging of the BCR/ABL fusion gene isoforms (e.g., e13a2, e14a2 and co-expression type) of chronic myeloid leukemia (CML) is of vital importance to first-line drug selection, but there is no assay that meets clinical needs (e.g., clinical kits > 18 h without isoforms information). Herein, an in situ imaging platform is developed for the rapid and accurate detection of CML fusion gene isoforms using asymmetric sequence-enhanced hairpins DNA encapsulated silver nanoclusters (ADHA) and catalyzed hairpin assembly (CHA). The specific detection of e13a2 and e14a2 fusion gene isoforms with detection limits of 19.2 am (11.558 copies µL-1 ) and 32.56 am (19.601 copies µL-1 ) in one-pot is achieved. The feasibility of the developed assay for real-world applications are demonstrated by one-step fluorescence imaging (40 min) of e13a2, e14a2 and co-expression type in bone marrow quantitatively (International Standard: 15.66%-168.878%) and further validated by cDNA-sequencing. This work suggests that the developed imaging platform holds great potential for rapid identification of the fusion gene isoforms and isoform related treatment monitoring.
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Proteínas de Fusão bcr-abl , Leucemia Mielogênica Crônica BCR-ABL Positiva , Humanos , Proteínas de Fusão bcr-abl/genética , Proteínas de Fusão bcr-abl/uso terapêutico , Medula Óssea , Prata/uso terapêutico , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Isoformas de Proteínas/genética , DNA Complementar , Imagem ÓpticaRESUMO
Salmonella typhimurium (S. typhimurium) and Staphylococcus aureus (S. aureus) are common food-borne pahogens that cause food poisoning in humans. In this study, we developed a method for the simultaneous determination of S. typhimurium and S. aureus based on multiplex polymerase spiral reaction (m-PSR) and melting curve analysis. Two pairs of primers were designed specifically to target the conserved invA gene of S. typhimurium and nuc gene of S. aureus, and the nucleic acid amplification reaction was achieved under isothermal conditions in the same reaction tube for 40 min at 61 °C, melting curve analysis of the amplification product was carried out. The distinct mean melting temperature allowed simultaneous differentiation of the two target bacteria in the m-PSR assay. The limit of detection of S. typhimurium and S. aureus that could be detected simultaneously was 4.1 × 10-4 ng genomic DNA and 2 × 101 CFU/mL pure bacterial culture. Based on this method, analysis of artificially contaminated samples showed excellent sensitivity and specificity consistent with those of pure bacterial cultures. This method is rapid, simultaneous and promises to be a useful tool for the detection of food-borne pathogens in the food industry.
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Salmonella typhimurium , Staphylococcus aureus , Humanos , Salmonella typhimurium/genética , Staphylococcus aureus/genética , Microbiologia de Alimentos , Técnicas de Amplificação de Ácido Nucleico , Sensibilidade e EspecificidadeRESUMO
Based on DNA strand replacement reaction and aptamer-specific recognition, a simple dual-labeled DNA nanostructure is designed for the simultaneous detection of Ochratoxin A (OTA) and aflatoxin B1 (AFB1). C1 is labeled with Cy3 and Cy5, while C2 and C3 are labeled with BHQ2. The fluorescence intensity of DNA nanostructure composed of C1, C2 and C3 is weak because of fluorescence resonance energy transfer. When OTA Aptamer (OTA-Apt) and AFB1 Aptamer (AFB1-Apt) are added to the homogeneous system at the same time, C1 can be replaced with the help of toehold strand displacement, resulting in fluorescence enhancement. In the presence of both OTA and AFB1, the toehold strand displacement reaction is inhibited due to preferential binding between the target and their corresponding aptamers. The limit of detection of OTA was 0.007 ng/mL and that of AFB1 was 0.03 ng/mL. The recoveries of OTA and AFB1 were 96%-101% and 97%-101% in the corn sample, and 99%-101% and 92%-106% in the wine sample. Compared with other sensors, the preparation of this aptasensor needs simpler experimental steps and a shorter total-preparing time, confirming the convenient, rapid, and time-saving operation process.
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Nanoestruturas , Aflatoxina B1/química , DNA/química , Corantes Fluorescentes/química , Limite de DetecçãoRESUMO
MicroRNAs (miRNAs) are a class of small, single-stranded non-coding RNAs which have critical functions in various biological processes. Increasing evidence suggested that abnormal miRNA expression was closely related to many human diseases, and they are projected to be very promising biomarkers for non-invasive diagnosis. Multiplex detection of aberrant miRNAs has great advantages including improved detection efficiency and enhanced diagnostic precision. Traditional miRNA detection methods do not meet the requirements of high sensitivity or multiplexing. Some new techniques have opened novel paths to solve analytical challenges of multiple miRNA detection. Herein, we give a critical overview of the current multiplex strategies for the simultaneous detection of miRNAs from the perspective of two different signal differentiation models, including label differentiation and space differentiation. Meanwhile, recent advances of signal amplification strategies integrated into multiplex miRNA methods are also discussed. We hope this review provides the reader with future perspectives on multiplex miRNA strategies in biochemical research and clinical diagnostics.
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MicroRNAs , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , BiomarcadoresRESUMO
Herein, a dual-signal output fluorescent aptamer sensor was constructed for the simultaneous detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA) using the specific recognition ability of aptamers and the programmability of DNA. A functional capture probe (cDNA) was designed with the black hole quenching motif BHQ1 labeled at the 5' end and biotin (bio) labeled at the 3' end. The fluorescent dye Cy3-labeled aflatoxin B1 aptamer (AFB1-Apt) and the carboxyfluorescein FAM-labeled ochratoxin A aptamer (OTA-Apt) were used as two fluorescent probes. The cDNA is anchored to the quenching material gold nanoflowers (AuNFs) by the action of streptavidin (SA) and biotin. Its ends can be complementarily paired with two fluorescent probe bases to form a double-stranded structure. The fluorescence of Cy3 was quenched by AuNFs, and the fluorescence of FAM was quenched by BHQ1 through the fluorescence energy resonance transfer (FRET) effect, forming a fluorescence quenching system. Due to the high affinity of the target and the aptamer, the structure of the aptamer probe changes and detaches from the sensor when AFB1 and OTA are present, resulting in enhanced fluorescence. Under optimal conditions, the linear range of AFB1 was 0.1-100 ng/mL (R2 = 0.996), the limit of detection (LOD) was as low as 0.014 ng/mL, and the limit of quantification (LOQ) was 0.046 ng/mL. The linear range of OTA was 0.1-100 ng/mL (R2 = 0.995), the limit of detection (LOD) was as low as 0.027 ng/mL, and the limit of quantification (LOQ) was 0.089 ng/mL. The sensor had high accuracy in detecting both AFB1 and OTA in real sample analysis. The results of the t test show that there is no significant difference between the results of this study and the high-performance liquid phase (HPLC) method, indicating that the prepared sensor can be used as a potential platform for multiple mycotoxins detection.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Micotoxinas , Micotoxinas/análise , DNA Complementar/química , Biotina , Ouro/química , Aptâmeros de Nucleotídeos/química , Corantes Fluorescentes/química , Aflatoxina B1/análise , Limite de DetecçãoRESUMO
Heavy metal ion (HMI) sensors are the most sought commercial devices for environmental monitoring and food analysis research due to serious health concerns associated with HMI overdosage. Herein, we developed an effective electrochemical sensor for simultaneous detection of four HMI (Cd2+, Pb2+, Fe2+, and Cu2+) using a ternary nanocomposite of reduced graphene oxide functionalized with polydopamine and alanine (ALA/pDA/rGO). Comprehensive spectroscopic and microscopic characterizations were performed to ensure the formation of the ternary nanocomposite. The developed nanocomposite on glassy carbon electrode (GCE) yields >2-fold higher current than GO/GCE electrode with excellent electrochemical stability and charge transfer rate. Using DPV, various chemical and electrochemical parameters, such as supporting electrolyte, buffer pH, metal deposition time, and potential, were optimized to achieve highly sensitive detection of targeted HMI. For Cd2+, Pb2+, Fe2+, and Cu2+ sensing devised sensor exhibited detection limits of 1.46, 2.86, 50.23, and 17.95 ppb and sensitivity of 0.0929, 0.0744, 0.0051, and 0.0394 µA/ppb, respectively, with <6% interference. The sensor worked similarly well for real water samples with HMI. This study demonstrates a novel strategy for concurrently detecting and quantifying multiple HMI in water and soil using a smart ternary nanocomposite-based electrochemical sensor, which can also detect HMI in food samples.
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Cádmio , Nanocompostos , Cádmio/análise , Chumbo , Óxidos/química , Técnicas Eletroquímicas/métodos , Carbono/química , Água , Nanocompostos/químicaRESUMO
Recently, it has been suggested that the mnemonic information that underlies recognition decisions changes when participants are asked to indicate whether a test stimulus is new rather than old (Brainerd et al., 2021, Journal of Experimental Psychology: Learning Memory, and Cognition, advance online publication). However, some observations that have been interpreted as evidence for this assertion need not be due to mnemonic changes, but may instead be the result of conservative response strategies if the possibility of asymmetric receiver operating characteristics (ROCs) is taken into account. Conversely, recent findings in support of asymmetric ROCs rely on the assumption that the mnemonic information accessed by the decision-maker does not depend on whether an old or a new item is considered to be the target Kellen et al. (2021, Psychological Review 128[6], 1022-1050). Here, we aim to clarify whether there is such a difference in accessibility of mnemonic information by applying signal detection theory. To this end, we used two versions of a simultaneous detection and identification task in which we presented participants with two test stimuli at a time. In one version, the old item was the target; in the other, the new item was the target. This allowed us to assess differences in mnemonic information retrieved in the two tasks while taking possible ROC asymmetry into account. Results clearly indicate that there is indeed a difference in the accessibility of mnemonic information as postulated by (Brainerd et al., 2021, Journal of Experimental Psychology: Learning Memory, and Cognition, advance online publication).
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Memória , Detecção de Sinal Psicológico , Humanos , Reconhecimento Psicológico/fisiologia , Cognição , Curva ROCRESUMO
Respiratory viruses usually induced similar clinical symptoms at early infection. Herein, we presented a multichannel surface-enhanced Raman scattering-based lateral flow immunoassay (SERS-based LFA) using high-performance magnetic SERS tags for the simultaneous ultrasensitive detection of respiratory viruses, namely influenza A virus (H1N1), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and respiratory syncytial virus (RSV) in biological samples. As-prepared magnetic SERS tags can directly enrich and capture target viruses without pretreatment of samples, avoiding the interference of impurities in the samples as well as improving the sensitivity. With the capture-detection method, the detection limits of the proposed assay reached 85 copies mL-1, 8 pg mL-1, and 8 pg mL-1 for H1N1, SARS-CoV-2 and RSV, respectively. Moreover, the detection properties of the proposed method for target viruses in throat swab samples were verified, suggesting its remarkable potential for the early and rapid differential diagnosis of respiratory viruses.
Assuntos
COVID-19 , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A , Humanos , Vírus Sinciciais Respiratórios , SARS-CoV-2 , COVID-19/diagnóstico , Fenômenos MagnéticosRESUMO
Seniors face many challenges as they age, such as dementia, cognitive and memory disorders, vision and hearing impairment, among others. Although most of them would like to stay in their own homes, as they feel comfortable and safe, in some cases, older people are taken to special institutions, such as nursing homes. In order to provide serious and quality care to elderly people at home, continuous remote monitoring is perceived as a solution to keep them connected to healthcare service providers. The new trend in medical health services, in general, is to move from 'hospital-centric' services to 'home-centric' services with the aim of reducing the costs of medical treatments and improving the recovery experience of patients, among other benefits for both patients and medical centers. Smart energy data captured from electrical home appliance sensors open a new opportunity for remote healthcare monitoring, linking the patient's health-state/health-condition with routine behaviors and activities over time. It is known that deviation from the normal routine can indicate abnormal conditions such as sleep disturbance, confusion, or memory problems. This work proposes the development and deployment of a smart energy data with activity recognition (SEDAR) system that uses machine learning (ML) techniques to identify appliance usage and behavior patterns oriented to older people living alone. The proposed system opens the door to a range of applications that go beyond healthcare, such as energy management strategies, load balancing techniques, and appliance-specific optimizations. This solution impacts on the massive adoption of telehealth in third-world economies where access to smart meters is still limited.