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1.
Anal Chem ; 96(1): 220-228, 2024 01 09.
Artículo en Inglés | MEDLINE | ID: mdl-38109169

RESUMEN

Nucleic acid detection of pathogens in a point-of-need (PON) manner is of great significance yet remains challenging for sensitive and accurate visual discrimination. Here, we report a CRISPR-Cas12a-mediated lateral flow assay for PON detection of Salmonella typhimurium (S.ty) that is a prevailing pathogen disseminated through tainted food. The variation of the fluorescence color of the test line is exploited to interpret the results, enabling the discrimination between positive and negative samples on the basis of a hue-recognition mechanism. By leveraging the cleavage activity of Cas12a and hue-recognition readout, the assay facilitated by recombinase polymerase amplification can yield a visual detection limit of 1 copy µL-1 for S.ty genomic DNA within 1 h. The assay also displays a high specificity toward S.ty in fresh chicken samples, as well as a sensitivity 10-fold better than that of the commercial test strip. Moreover, a semiquantitative detection of S.ty ranging from 0 to 4 × 103 CFU/mL by the naked eye is made possible, thanks to the easily discernible color change of the test line. This approach provides an easy, rapid, accurate, and user-friendly solution for the PON detection of Salmonella and other pathogens.


Asunto(s)
Sistemas CRISPR-Cas , Salmonella typhimurium , Animales , Sistemas CRISPR-Cas/genética , Salmonella typhimurium/genética , Bioensayo , Pollos , Alimentos , Técnicas de Amplificación de Ácido Nucleico
2.
BMC Public Health ; 24(1): 414, 2024 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-38331772

RESUMEN

IMPORTANCE: Contact tracing is the process of identifying people who have recently been in contact with someone diagnosed with an infectious disease. During an outbreak, data collected from contact tracing can inform interventions to reduce the spread of infectious diseases. Understanding factors associated with completion rates of contact tracing surveys can help design improved interview protocols for ongoing and future programs. OBJECTIVE: To identify factors associated with completion rates of COVID-19 contact tracing surveys in New York City (NYC) and evaluate the utility of a predictive model to improve completion rates, we analyze laboratory-confirmed and probable COVID-19 cases and their self-reported contacts in NYC from October 1st 2020 to May 10th 2021. METHODS: We analyzed 742,807 case investigation calls made during the study period. Using a log-binomial regression model, we examined the impact of age, time of day of phone call, and zip code-level demographic and socioeconomic factors on interview completion rates. We further developed a random forest model to predict the best phone call time and performed a counterfactual analysis to evaluate the change of completion rates if the predicative model were used. RESULTS: The percentage of contact tracing surveys that were completed was 79.4%, with substantial variations across ZIP code areas. Using a log-binomial regression model, we found that the age of index case (an individual who has tested positive through PCR or antigen testing and is thus subjected to a case investigation) had a significant effect on the completion of case investigation - compared with young adults (the reference group,24 years old < age < = 65 years old), the completion rate for seniors (age > 65 years old) were lower by 12.1% (95%CI: 11.1% - 13.3%), and the completion rate for youth group (age < = 24 years old) were lower by 1.6% (95%CI: 0.6% -2.6%). In addition, phone calls made from 6 to 9 pm had a 4.1% (95% CI: 1.8% - 6.3%) higher completion rate compared with the reference group of phone calls attempted from 12 and 3 pm. We further used a random forest algorithm to assess its potential utility for selecting the time of day of phone call. In counterfactual simulations, the overall completion rate in NYC was marginally improved by 1.2%; however, certain ZIP code areas had improvements up to 7.8%. CONCLUSION: These findings suggest that age and time of day of phone call were associated with completion rates of case investigations. It is possible to develop predictive models to estimate better phone call time for improving completion rates in certain communities.


Asunto(s)
COVID-19 , Adolescente , Adulto Joven , Humanos , Adulto , Anciano , COVID-19/epidemiología , Trazado de Contacto/métodos , Ciudad de Nueva York/epidemiología , Encuestas y Cuestionarios , Brotes de Enfermedades
3.
Anal Chem ; 94(50): 17567-17576, 2022 12 20.
Artículo en Inglés | MEDLINE | ID: mdl-36458677

RESUMEN

The development of highly specific biomimetic recognition material is a challenge for rapid detection of harmful residues in foodstuff. In this study, a paper-based boronate affinity metal-organic framework/molecularly imprinted polymer microfluidic chip (FZS-BA@MIP) was constructed based on the in situ construction strategy, which was also designed as a highly specific biomimetic recognition module. Here, the homogeneous zeolitic imidazole framework-8 (ZIF-8) membrane served as a great scaffold and enrichment layer. Besides, the recognition layer of MIP was prepared based on a highly oriented boronate affinity surface imprinting strategy. With the aid of the liquid flow channel, the highly specific enrichment and visual detection for antibiotic residues like kanamycin in actual products were achieved on the paper chip module of an integrated lateral flow platform. The whole analysis process could be accomplished within 30 min. In brief, this study offered a new integrated biomimetic recognition platform for visually detecting harmful veterinary residues containing cis-diols, which demonstrated promising commercial value in point-of-care testing of foodborne hazardous compounds.


Asunto(s)
Materiales Biomiméticos , Impresión Molecular , Biomimética , Kanamicina , Materiales Biomiméticos/química
4.
Crit Rev Food Sci Nutr ; 62(28): 7849-7865, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-33955294

RESUMEN

Aflatoxins (AFs) contamination in food and agricultural products poses a significant threat to human health. Sensitive and accurate detection of AFs provides a strong guarantee for ensuring food safety. Conventional chromatographic-based or mass spectrum methods, which rely on bulky instrument and skilled personnel, are not suitable for on-site surveillance. By contrast, visual detections which possess the merits of rapidity and sophisticated instrument-free present an excellent potential for the on-site detection of AFs. This review intends to summarize the latest development of visual methods for AFs detection, including paper-based tests, chromogenic reactions, and luminescent methods. Emerging technologies, like nanotechnology, DNAzymes, and aptamers combined with these visual methods are introduced. The basic principles, features, and application advantages of each type of visual methods are discussed. The biggest challenges and perspectives on their future trends are also addressed.


Asunto(s)
Aflatoxinas , ADN Catalítico , Aflatoxinas/análisis , Contaminación de Alimentos/análisis , Inocuidad de los Alimentos , Humanos , Nanotecnología
5.
Anal Chem ; 93(42): 14300-14306, 2021 10 26.
Artículo en Inglés | MEDLINE | ID: mdl-34645259

RESUMEN

Vibrio parahaemolyticus (V. parahaemolyticus), which may cause gastrointestinal disorders in humans, is a pathogen commonly found in seafood. There are many methods for detecting V. parahaemolyticus, yet they have some shortcomings, such as high cost, labor-intensiveness, and complicated operation, which are impractical for resource-limited settings. Herein, we present a sequence-specific, label-free, and colorimetric method for visual detection of V. parahaemolyticus. This method utilizes CRISPR/Cas12a to specifically recognize the loop-mediated isothermal amplification (LAMP) products for further trans-cleaving the G-quadruplex DNAzyme and depriving its peroxidase-mimicking activity. In this way, the results can be directly observed with the naked eyes via the color development of 2,2'-azino-di-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS2-), which displays colorless for positive samples while green for target-free samples. We term such Cas12a-crRNA preventing ABTS2- from developing color by trimming the G-quadruplex DNAzyme as Cascade. The proposed method can detect 9.8 CFU (per reaction) of pure cultured V. parahaemolyticus, and the sensitivity is comparable to real-time LAMP. It has been applied for practical use and showed the capability to detect 6.1 × 102 CFU/mL V. parahaemolyticus in shrimp samples. Based on this, the newly established Cascade method can be employed as a universal biosensing strategy for pathogenic bacterial testing in the field.


Asunto(s)
ADN Catalítico , Vibrio parahaemolyticus , Sistemas CRISPR-Cas , Colorimetría , Humanos , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Vibrio parahaemolyticus/genética
6.
Crit Rev Biochem Mol Biol ; 50(6): 477-88, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26362261

RESUMEN

A dedicated network of cellular factors ensures that proteins translocated into the endoplasmic reticulum (ER) are folded correctly before they exit this compartment en route to other cellular destinations or for secretion. When proteins misfold, selective ER-resident enzymes and chaperones are recruited to rectify the protein-misfolding problem in order to maintain cellular proteostasis. However, when a protein becomes terminally misfolded, it is ejected into the cytosol and degraded by the proteasome via a pathway called ER-associated degradation (ERAD). Strikingly, toxins and viruses can hijack elements of the ERAD pathway to access the host cytosol and cause infection. This review focuses on emerging data illuminating the molecular mechanisms by which these toxic agents co-opt the ER-to-cytosol translocation process to cause disease.


Asunto(s)
Infecciones Bacterianas/metabolismo , Fenómenos Fisiológicos Bacterianos , Toxinas Bacterianas/metabolismo , Degradación Asociada con el Retículo Endoplásmico , Interacciones Huésped-Patógeno , Infecciones por Polyomavirus/metabolismo , Poliomavirus/fisiología , Animales , Citosol/metabolismo , Citosol/microbiología , Citosol/virología , Retículo Endoplásmico/metabolismo , Retículo Endoplásmico/microbiología , Retículo Endoplásmico/virología , Humanos , Transporte de Proteínas
7.
Anal Chem ; 87(19): 9998-10005, 2015 Oct 06.
Artículo en Inglés | MEDLINE | ID: mdl-26358143

RESUMEN

Near-infrared (NIR) quantum dots (QDs) have emerged as an attractive bioimaging toolkit for exploring biological events because they can provide deep imaging penetration and low fluorescence background. However, the quantitation process of such NIR QDs generally relies on single-emission intensity change, which is susceptible to a variety of environmental factors. Herein, for the first time, we proposed a protein-directed co-template strategy to synthesize a NIR-based, dual-emission fluorescent nanohybrid (DEFN) constructed from far-red gold nanoclusters and NIR PbS QDs (AuNCs-PbS-QDs). The convenient protein-directed co-template synthesis avoids the tedious chemical coupling and modification required in conventional preparation approaches of DEFNs. Additionally, the dual-emission signals of AuNCs-PbS-QDs exhibit two well-resolved emission peaks (640 and 813 nm) separated by 173 nm, which can eliminate environmental interferences by the built-in correction of ratiometric signal, resulting in a more favorable system for bioimaging and biosensing. Next, the target-responsive capability of this NIR-based DEFN to ascorbic acid (AA) was discovered, enabling the proposed DEFN to ratiometrically detect AA with a linear range of 3-40 µM and a detection limit of 1.5 µM. This DEFN sensor possesses high selectivity, rapid response, and excellent photostability. Moreover, the feasibility of this NIR nanosensor has been fully proved by the ratiometric detection of AA for fruit internal quality assessment, in vitro cellular imaging, and in vivo imaging in nude mice.


Asunto(s)
Ácido Ascórbico/análisis , Colorantes Fluorescentes/química , Oro/química , Plomo/química , Nanopartículas del Metal/química , Puntos Cuánticos/química , Sulfuros/química , Animales , Células HeLa , Humanos , Límite de Detección , Nanopartículas del Metal/ultraestructura , Ratones , Ratones Desnudos , Imagen Óptica , Puntos Cuánticos/ultraestructura
8.
Toxicol Appl Pharmacol ; 278(2): 107-15, 2014 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-24793808

RESUMEN

The foodborne mycotoxin deoxynivalenol (DON) induces a ribotoxic stress response in mononuclear phagocytes that mediate aberrant multi-organ upregulation of TNF-α, interleukins and chemokines in experimental animals. While other DON congeners also exist as food contaminants or pharmacologically-active derivatives, it is not known how these compounds affect expression of these cytokine genes in vivo. To address this gap, we compared in mice the acute effects of oral DON exposure to that of seven relevant congeners on splenic expression of representative cytokine mRNAs after 2 and 6h. Congeners included the 8-ketotrichothecenes 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX), nivalenol (NIV), the plant metabolite DON-3-glucoside (D3G) and two synthetic DON derivatives with novel satiety-inducing properties (EN139528 and EN139544). DON markedly induced transient upregulation of TNF-α IL-1ß, IL-6, CXCL-2, CCL-2 and CCL-7 mRNA expressions. The two ADONs also evoked mRNA expression of these genes but to a relatively lesser extent. FX induced more persistent responses than the other DON congeners and, compared to DON, was: 1) more potent in inducing IL-1ß mRNA, 2) approximately equipotent in the induction of TNF-α and CCL-2 mRNAs, and 3) less potent at upregulating IL-6, CXCL-2, and CCL-2 mRNAs. EN139528's effects were similar to NIV, the least potent 8-ketotrichothecene, while D3G and EN139544 were largely incapable of eliciting cytokine or chemokine mRNA responses. Taken together, the results presented herein provide important new insights into the potential of naturally-occurring and synthetic DON congeners to elicit aberrant mRNA upregulation of cytokines associated with acute and chronic trichothecene toxicity.


Asunto(s)
Quimiocinas/biosíntesis , Citocinas/biosíntesis , Regulación de la Expresión Génica , Mediadores de Inflamación/metabolismo , ARN Mensajero/biosíntesis , Tricotecenos/administración & dosificación , Tricotecenos/síntesis química , Regulación hacia Arriba , Administración Oral , Animales , Quimiocinas/agonistas , Quimiocinas/genética , Citocinas/agonistas , Citocinas/genética , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Mediadores de Inflamación/agonistas , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , ARN Mensajero/agonistas , Resultado del Tratamiento , Regulación hacia Arriba/genética , Regulación hacia Arriba/inmunología
9.
ACS Sens ; 2024 Apr 23.
Artículo en Inglés | MEDLINE | ID: mdl-38651662

RESUMEN

Excavating nucleic acid quantitative capabilities by combining clustered regularly interspaced short palindromic repeats (CRISPR) and isothermal amplification in one pot is of common interest. However, the mutual interference between CRISPR cleavage and isothermal amplification is the primary obstacle to quantitative detection. Though several works have demonstrated enhanced detection sensitivity by reducing the inhibition of CRISPR on amplification in one pot, few paid attention to the amplification process and even dynamic reaction processes between the two. Herein, we find that DNA quantification can be realized by regulating either recombinase polymerase amplification (RPA) efficiency or CRISPR/Cas12a cleaving efficiency (namely, tuning the dynamic reaction balance) in one pot. The sensitive quantification is realized by utilizing dual PAM-free crRNAs for CRISPR/Cas12a recognition. The varied RPA primer concentration with stabilized CRISPR systems significantly affects the amplification efficiency and quantitative performances. Alternatively, quantitative detection can also be achieved by stabilizing the amplification process while regulating the CRISPR/Cas12a concentration. The quantitative capability is proved by detecting DNA targets from Lactobacillus acetotolerans and SARS-CoV-2. The quantitative performance toward real samples is comparable to quantitative real-time PCR for detecting L. acetotolerans spiked in fermented food samples and SARS-CoV-2 clinical samples. We expect that the presented method will be a powerful tool for quantifying other nucleic acid targets.

10.
Anal Chem ; 85(13): 6279-86, 2013 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-23742234

RESUMEN

In this paper, the efficient quenching effect of deoxyguanosine-5'-phosphate (dGMP) on anodic electrochemiluminescence (ECL) of the CdTe/ZnS quantum dots (QDs) is reported for the first time. This ECL quenching was found to be specific for free dGMP and not observed for dGMP residues in different DNA structures. The unique dGMP-based QDs ECL quenching was then utilized to develop a versatile biosensing strategy to determine various protein-DNA interactions with the assistance of exonuclease, Exo I, to hydrolyze DNA and liberate dGMP. Taking single-stranded DNA binding protein (SSB) and thrombin as examples, two novel detection modes have been developed based on dGMP-QDs ECL strategy. The first method used hairpin probes and SSB-promoted probe cleavage by Exo I for facile signal-off detection of SSB, with a wide linear range of 1-200 nM and a low detection limit of 0.1 nM. The second method exploited aptamer-thrombin binding to protect probes against Exo I degradation for sensitive signal-on detection of thrombin, giving a linear response over a range of 1-150 nM and a detection limit as low as 0.1 nM. Both methods were homogeneous and label-free without QDs or DNA modification. Therefore, this dGMP-specific QDs ECL quenching presents a promising detection mechanism suitable for probing various protein-nucleic acid interactions.


Asunto(s)
Técnicas Biosensibles/métodos , Nucleótidos de Desoxiguanina/análisis , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Puntos Cuánticos/química , Trombina/análisis , Compuestos de Cadmio/química , Nucleótidos de Desoxiguanina/metabolismo , Ácidos Nucleicos/análisis , Ácidos Nucleicos/metabolismo , Unión Proteica/fisiología , Telurio/química , Trombina/metabolismo , Sulfato de Zinc/química
11.
Talanta ; 251: 123793, 2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-35952501

RESUMEN

Circulating tumor cells (CTCs), as a type of tumor, have attracted wide attention because of their characteristics of shedding from the primary tumor and spreading to other tissues and organs through peripheral blood. The circulating tumor DNA (ctDNA), the DNA released by CTCs and other tumor cells into the peripheral blood, was considered as a promising detection substance for clinical application. By utilizing the biocompatibility of red blood cells to realize the attachment of tetrahedral DNA (TDN), as well as the specific target recognition ability of TDN to enable efficient recognition of targets, a biocompatible electrochemical biosensor for effective and rapid detection of ctDNA was developed using methylene blue (MB) as the signal probe. The current signal and the logarithm of ctDNA concentration were linearly correlated in the range from 1 fM to 100 pM with the detection limit of 0.66 fM. With high specificity, the TDN-based biosensor can detect ctDNA efficiently in the real biological environment such as serum, which provided a potential opportunity for the early clinical diagnosis.


Asunto(s)
Técnicas Biosensibles , ADN Tumoral Circulante , Nanoestructuras , Células Neoplásicas Circulantes , ADN/química , Técnicas Electroquímicas , Eritrocitos , Humanos , Límite de Detección , Azul de Metileno , Nanoestructuras/química
12.
Biosensors (Basel) ; 13(5)2023 Apr 26.
Artículo en Inglés | MEDLINE | ID: mdl-37232864

RESUMEN

Salmonella is one of four key global causes of diarrhea, and in humans, it is generally contracted through the consumption of contaminated food. It is necessary to develop an accurate, simple, and rapid method to monitor Salmonella in the early phase. Herein, we developed a sequence-specific visualization method based on loop-mediated isothermal amplification (LAMP) for the detection of Salmonella in milk. With restriction endonuclease and nicking endonuclease, amplicons were produced into single-stranded triggers, which further promoted the generation of a G-quadruplex by a DNA machine. The G-quadruplex DNAzyme possesses peroxidase-like activity and catalyzes the color development of 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) as the readouts. The feasibility for real samples analysis was also confirmed with Salmonella spiked milk, and the sensitivity was 800 CFU/mL when observed with the naked eye. Using this method, the detection of Salmonella in milk can be completed within 1.5 h. Without the involvement of any sophisticated instrument, this specific colorimetric method can be a useful tool in resource-limited areas.


Asunto(s)
ADN Catalítico , G-Cuádruplex , Humanos , ADN Catalítico/genética , ADN , Salmonella/genética , Técnicas de Amplificación de Ácido Nucleico/métodos
13.
RSC Adv ; 13(22): 14776-14781, 2023 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-37197180

RESUMEN

A novel covalent organic framework (COF) (Tp-BI-COF) with combined ketimine-type enol-imine and keto-enamine linkages was prepared through a cascade of ketimine condensation followed by aldimine condensation and characterized by XRD, solid state 13C NMR, IR, TGA and BET. Tp-BI-COF showed high stability toward acid, organic solvent, and boiling water. The 2D COF exhibited photochromic properties after being irradiated with a xenon lamp. The stable COF, with aligned one-dimensional nanochannels, provided nitrogen sites on pore walls, which confine and stabilize the H3PO4 in the channel via hydrogen-bonding interactions. After loading with H3PO4, the material showed excellent anhydrous proton conductivity.

14.
Toxicol Appl Pharmacol ; 265(1): 10-8, 2012 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-23022514

RESUMEN

The Type B trichothecene deoxynivalenol (DON), a ribotoxic mycotoxin known to contaminate cereal-based foods, induces ribosomal RNA (rRNA) cleavage in the macrophage via p38-directed activation of caspases. Here we employed the RAW 264.7 murine macrophage model to test the hypothesis that this rRNA cleavage pathway is similarly induced by other ribotoxins. Capillary electrophoresis confirmed that the antibiotic anisomycin (≥25ng/ml), the macrocylic trichothecene satratoxin G (SG) (≥10ng/ml) and ribosome-inactivating protein ricin (≥300ng/ml) induced 18s and 28s rRNA fragmentation patterns identical to that observed for DON. Also, as found for DON, inhibition of p38, double-stranded RNA-activated kinase (PKR) and hematopoietic cell kinase (Hck) suppressed MAPK anisomycin-induced rRNA cleavage, while, in contrast, their inhibition did not affect SG- and ricin-induced rRNA fragmentation. The p53 inhibitor pifithrin-µ and pan caspase inhibitor Z-VAD-FMK suppressed rRNA cleavage induced by anisomycin, SG and ricin, indicating that these ribotoxins shared with DON a conserved downstream pathway. Activation of caspases 8, 9 and 3 concurrently with apoptosis further suggested that rRNA cleavage occurred in parallel with both extrinsic and intrinsic pathways of programmed cell death. When specific inhibitors of cathepsins L and B (lysosomal cysteine cathepsins active at cytosolic neutral pH) were tested, only the former impaired anisomycin-, SG-, ricin- and DON-induced rRNA cleavage. Taken together, the data suggest that (1) all four ribotoxins induced p53-dependent rRNA cleavage via activation of cathepsin L and caspase 3, and (2) activation of p53 by DON and anisomycin involved p38 whereas SG and ricin activated p53 by an alternative mechanism.


Asunto(s)
División del ARN/efectos de los fármacos , ARN Ribosómico/efectos de los fármacos , Tricotecenos/toxicidad , Animales , Anisomicina/toxicidad , Apoptosis/efectos de los fármacos , Western Blotting , Caspasa 8/efectos de los fármacos , Catepsina L/farmacología , Células Cultivadas , Activación Enzimática/efectos de los fármacos , Ratones , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Inhibidores de la Síntesis del Ácido Nucleico/toxicidad , Proteínas Proto-Oncogénicas c-hck/metabolismo , ARN Ribosómico/aislamiento & purificación , Ricina/toxicidad , Tricotecenos/farmacología , Proteína p53 Supresora de Tumor/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo
15.
Chemistry ; 18(13): 3992-9, 2012 Mar 26.
Artículo en Inglés | MEDLINE | ID: mdl-22337073

RESUMEN

The DNA nick repair catalyzed by DNA ligase is significant for fundamental life processes, such as the replication, repair, and recombination of nucleic acids. Here, we have employed ligase to regulate DNAzyme activity and developed a homogeneous, colorimetric, label-free and DNAzyme-based strategy to detect DNA ligase activity. This novel strategy relies on the ligation-trigged activation or production of horseradish peroxidase mimicking DNAzyme that catalyzes the generation of a color change signal; this results in a colorimetric assay of DNA ligase activity. Using T4 DNA ligase as a model, we have proposed two approaches to demonstrate the validity of the DNAzyme strategy. The first approach utilizes an allosteric hairpin-DNAzyme probe specifically responsive to DNA ligation; this approach has a wide detection range from 0.2 to 40 U mL(-1) and a detection limit of 0.2 U mL(-1). Furthermore, the approach was adapted to probe nucleic acid phosphorylation and single nucleotide mismatch. The second approach employs a "split DNA machine" to produce numerous DNAzymes after being reassembled by DNA ligase; this greatly enhances the detection sensitivity by a signal amplification cascade to achieve a detection limit of 0.01 U mL(-1).


Asunto(s)
Colorimetría/métodos , ADN Ligasas/metabolismo , ADN Catalítico/metabolismo , ADN/química , ADN Ligasa (ATP) , ADN Ligasas/análisis , Peroxidasa de Rábano Silvestre/metabolismo , Modelos Moleculares , Conformación de Ácido Nucleico , Hibridación de Ácido Nucleico
16.
Talanta ; 243: 123292, 2022 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-35202837

RESUMEN

Antibiotics are widely used for improving the living conditions of livestock. However, residual antibiotics present in animal products induce several human diseases. Therefore, a simple, rapid, and cost-effective system for detecting and monitoring the presence of antibiotics in foods is in great demand to alleviate safety concerns. In this study, a highly sensitive and selective aptameric electrochemical sensing platform was designed based on nanomaterial modification and DNA nanotechnology. Electrochemically reduced graphene oxide and gold nanoparticles were used to modify the working surface of a screen-printed electrode to enhance electrical conductivity and biocompatibility. The electrode surface was further modified with self-assembled tetrahedral DNA nanostructures (TDN) to improve the detection sensitivity. The TDN allowed controlling the nano-spacing of aptamers immobilized on the electrode surface and placing aptamers in a solution-phase-like detecting environment to improve the target-binding efficiency without signal amplification modules. Differential pulse voltammetry was employed to measure electrical signals in proportion to the amount of ampicillin, the target antibiotic, present in buffer and spiked milk samples. The designed aptasensor was able to detect and measure the target ampicillin in less than 30 min over a wide concentration range of 10 pM to 1 mM, with a limit of detection of 1 pM, which is 100 times better than when using the same sensing probe without TDN modification. The aptasensor was reusable by simply rinsing with deionized water, remained stable during 15-day storage, and yielded reproducible results.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Nanopartículas del Metal , Nanoestructuras , Ampicilina , Animales , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , ADN/química , Técnicas Electroquímicas/métodos , Oro/química , Límite de Detección , Nanopartículas del Metal/química
17.
J Hazard Mater ; 423(Pt A): 126962, 2022 02 05.
Artículo en Inglés | MEDLINE | ID: mdl-34464866

RESUMEN

Colorimetric and fluorescent methods for Ochratoxin A (OTA) detection are convenient and well received. However, the pigments and autofluorescence originated from food matrices often interfere with detection signals. We have developed a strategy with colorimetric and fluorescent dual modes to solve this challenge. In the colorimetric mode, OTA aptamer (AP9) was assembled into a DNA triple-helix switch with a specially designed signal-amplifying sequence. The OTA-induced G-quadruplex (G4) of AP9 would open the switch and release the signal-amplifying sequence for colorimetric signal amplification. The G4 structures of AP9 were further utilized to combine with the fluorogenic dye ThT for fluorescent mode. By skillfully engineering DNA G4 assembly for signal amplification, there was no need for any DNA amplification or nanomaterials labeling. Detections could be carried out in a wide temperature range (22-37 â„ƒ) and finished rapidly (colorimetric mode, 60 min; fluorescent mode, 15 min). Broad linear ranges (colorimetric mode, 10-1.5 ×103 µg/kg; fluorescent mode, 0.05-1.0 ×103 µg/kg) were achieved. The limit of detection for colorimetric and fluorescent modes were 4 µg/kg and 0.01 µg/kg, respectively. The two modes have been successfully applied to detect OTA in samples with intrinsic pigments and autofluorescence, showing their applicability and reliability.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Colorimetría , ADN , Límite de Detección , Ocratoxinas , Reproducibilidad de los Resultados
18.
J Hazard Mater ; 411: 125090, 2021 06 05.
Artículo en Inglés | MEDLINE | ID: mdl-33453667

RESUMEN

The convenient and effective detection of toxins is urgently demanded for food security and human health. Herein, based on the catalytic activity of mimetic peroxidase from the Cu2O@Fe(OH)3 yolk-shell nanocages, a dual-modal multi-colorimetric and ratiometric fluorescence immunosensor for the sensitive detection of ochratoxin A (OTA) was successfully developed. For the multi-colorimetric detection, H2O2 can be effectively decomposed by Cu2O@Fe(OH)3 to form ·OH groups, thus Au nanorods (Au NRs) can be etched to exhibit vivid color variations and localized surface plasmon resonance (LSPR) shifts. For the ratiometric fluorescence detection, o-phenylenediamine was oxidized by Cu2O@Fe(OH)3 to form 2,3-diaminophenazine (DAP) in the presence of H2O2. Interestingly, the exogenous fluorescence signal source of carbon dots can be quenched by DAP via inner filter effect, while a new emission peak at 563 nm can be discovered, forming a ratiometric fluorescence signal. Due to the independent signals and mutual confirmation, the performance of the dual-modal immunosensor for the detection of OTA was significantly improved, where a broad linear range from 1 ng/L to 10 µg/L with a detection limit of 0.56 ng/L (S/N = 3) was achieved. The sensing strategy was also used to monitor OTA in millet and lake water samples with a satisfied performance.


Asunto(s)
Técnicas Biosensibles , Colorimetría , Ocratoxinas/análisis , Biomimética , Peróxido de Hidrógeno , Inmunoensayo , Límite de Detección , Peroxidasa
19.
Biosens Bioelectron ; 169: 112602, 2020 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-32947078

RESUMEN

The rapid growth of in vitro nucleic acid replication has offered a powerful tool for clinical diagnosis, food safety detection and environmental monitorning. Successful implementation of various isothermal nucleic acid amplification methods enables rapid replication of target sequences without the participant of a thermal cycler. Point-of-need analysis possesses great superiorities in user-friendly, instant results analysis, low manufacturing, and consumable costs. To meet the great challenge of point-of-need analysis, developing simple and rapid visual methods becomes crucial. Submicro- and nanomaterials possess unique surface properties, which enables their rapid response to DNA amplicons. Their unique optical, magnetic, catalytic, and other physical/chemical properties have been frequently employed for the visual detection of in vitro nucleic acid replications. Herein, we aim to review the submicro- and nanomaterials-based visual methods for detection of nucleic acid amplification. The visual methods are classified according to the designing strategies (e.g. LSPR, bridging flocculation, luminescence, catalytic reaction, separation, etc.). The basic principles, merits and drawbacks of each strategy are described. The application in analysis of nucleic acid targets and non-nucleic acid targets are discussed. The main challenges and future research directions are also highlighted in this rapidly emerging field.


Asunto(s)
Técnicas Biosensibles , Ácidos Nucleicos , ADN/genética , Humanos , Magnetismo , Técnicas de Amplificación de Ácido Nucleico , Ácidos Nucleicos/genética
20.
Spectrochim Acta A Mol Biomol Spectrosc ; 235: 118266, 2020 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-32217441

RESUMEN

In this work, a smart analytical strategy that combines excitation-emission matrix (EEM) fluorescence detection with alternating trilinear decomposition (ATLD) algorithm was developed for fast, on-line and interference-free study on the photodegradation kinetics of aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1) in rice and wheat under UV-Vis light (λ = 250-500 nm) treatment. With the aid of prominent "second-order advantage" of ATLD method, pure fluorescence signals of two targeted analytes can be directly resolved out from heavily overlapping spectral environment and accurately quantified even in the presence of unknown matrix interferences. Cereal samples in kinetic processing of photodegradation were detected without complex pretreatment steps except for a simple extraction using methanol/water solution (4:1, v/v), which solves the problem facing varied matrix interferences in the case of on-line monitoring of aflatoxins. The kinetic signals of analytes of interest were directly extracted regardless of varied matrix backgrounds of various cereals. The kinetic curves and degradation speeds of AFB1 and AFG1 can be estimated by resolved quantitative data, optimal radiation conditions including 365 nm wavelength and 35 J m-2 density were discussed for high-efficiency detoxification control of aflatoxins in rice and wheat. This strategy was promising to be as an alternative tool for eco-friendly photodegradation kinetic study of mycotoxins or other hazards in complex foodstuff matrixes.


Asunto(s)
Aflatoxinas/análisis , Contaminación de Alimentos/análisis , Espectrometría de Fluorescencia , Algoritmos , Calibración , Cromatografía Líquida de Alta Presión , Grano Comestible , Fluorescencia , Cinética , Metanol/química , Modelos Teóricos , Oryza/química , Fotólisis , Programas Informáticos , Triticum/química , Agua/química
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