Mehmandoust, MohammadGumus, Z. PinarSoylak, MustafaErk, Nevin...
8页
查看更多>>摘要：A label-free electrochemical biosensing approach as an appropriate analysis technique for SARS-CoV-2 spike protein (SARS-CoV-2 S-protein) was investigated to facilitate the diagnosis of coronavirus in real samples. It is crucial to construct diagnostic features that can rapidly identify infected individuals to limit the spread of the virus and assign treatment choices. Therefore, a novel and selective method using SiO2@UiO-66 and a label-free electrochemical immunoassay for rapidly detecting spike protein. The development of innovative approaches for direct viral detection employing simplified and ideally reagent-free assays is a pressing and difficult topic. The absence of speedy and effective ways to diagnose viral diseases especially SARS-CoV-2 on demand has worsened the issue of combating the COVID-19 pandemic. The developed electrode illustrated a wide dynamic range of 100.0 fg mL(-1) to 10.0 ng mL(-1) with low limit detection. Therefore, the as-fabricated electrochemical SARS-CoV2 S-protein sensor suggests an appropriate perspective in the point-of-care system, within 5.0 min, in nasal samples with satisfactory recovery.
查看更多>>摘要：Nucleic acid amplification tests have been widely applied in clinical diagnostics, food safety monitoring, and molecular biology. As a well-established isothermal amplification method, Loop-mediated isothermal amplification (LAMP) has gained recognition. However, the need for specifically designed four to six primers and nonspecific amplification pose challenges for further application of LAMP based detection methods. Here, a novel isothermal amplification method, termed closed dumbbell mediated isothermal amplification (CDA) of nucleic acids, was developed. The primers are easily designed by adding two different parts of middle sequence to the canonical PCR primers at 5 '-ends. CDA method was demonstrated in detecting MERS-CoV orf1a gene and H1N1 gene fragments with merits of short core primer, simple primer design process and high amplification efficiency. In addition, CDA showed excellent amplification efficacy over LAMP and competitive annealing mediated isothermal amplification (CAMP) by slight modification of primers targeting at same sequence. Furthermore, real-time and HNB based colorimetric CDA detection of Shigella were developed for practical application, both exhibited 100% success. In all, the developed CDA method with high specificity, simplicity, efficiency and rapidity has shown its great potential for point of care nucleic acids diagnostic.
Islam, Md. FahamidulIslam, Md. TarikulHasan, Md. MahmudulRahman, Mohammed M....
11页
查看更多>>摘要：Nickel particles alone can oxidize hydrogen peroxide but confronts extreme stability problem which imparts a barrier to act as sensor. The porous Nafion bed on glassy carbon electrode (GCE) surface provides the sureness of incorporating of Ni particles which was further exploited as an electrochemical sensor for H2O2 detection through oxidative degradation process. The simple electrochemical incorporation of Ni particles along the pores of Nafion improves the stability of the sensor significantly. The oxidative pathway of hydrogen peroxide on GCE/ Nafion/Ni was probed by analyzing mass transfer dependent linear sweep voltammograms both in static and rotating modes along with chronoamperometry. An electron transfer step determines the overall reaction rate with k degrees = 2.72 x 10-4 cm s- 1, which is supported by the values of transfer coefficient (beta) in between (0.68-0.75). Sensing performance was evaluated by recording differential pulse voltammograms (DPVs) with the linear detection limit (LOD) of 1.8 mu M and linear dynamic range (LDR) of 5-500 mu M. Real samples from industrial sources were successfully quantified with excellent reproducibility mark GCE/Nafion/Ni electrode as an applicable sensor.
查看更多>>摘要：Saxitoxin (STX), is one of the most dangerous and widespread paralytic shellfish toxins, causing a severe threat to the ecosystem and human health. So, it is important and highly essential to develop novel techniques for STX detection in a convenient, desirable, and low-cost manner. Herein, this study developed an electrolyte-insulator semiconductor (EIS) sensor covered with a layer-by-layer prepared, positively-charged weak polyelectrolyte layer of poly (allylamine hydrochloride) (PAH) for the label-free detection of STX. The specific aptamer (Apt) sensitive to STX was electrostatically adsorbed onto the PAH layer. This leads to a preferentially flat orientation of the Apt within the Debye length, thus yielding a reduced charge-screening influence and a higher sensor signal. Each step of sensor surface modification, i.e. PAH adsorption, immobilization of Apt, and attachment of STX, was monitored by capacitance-voltage (C-V) and constant-capacitance (ConCap) measurements. Furthermore, atomic force microscopy (AFM) was employed to characterize the surface morphology and roughness of the PAH layer. Fluorescence microscopy was used to confirm the effective immobilization of Apt onto the PAHmodified EIS sensor. The results showed that the detection range of this aptasensor for STX detection was 0.5-100 nM and the detection limit was as low as 0.05 nM. Furthermore, this aptasensor showed good selectivity and 9 days' stability. The mussel tissue extraction test suggested that this aptasensor can be used to detect STX in real samples. This aptasensor provides a convenient approach for moderate, rapid, and label-free detection of marine biological toxins.
查看更多>>摘要：Covid-19 variants transmissibility was quantitatively analyzed in silico to understand the reaction mechanisms and to find the reaction inhibitors. Especially, SARS-CoV-2 omicron mutant (omicron S-RBD) binding affinity with human angiotensin-converting enzyme-2 (ACE-2) was quantitatively analyzed using molecular interaction (MI) energy values (kcal.mol- 1) between the S-RBD and ACE-2. The MI of their optimized complex structures demonstrated that omicron's MI value (749.8) was 1.4 times delta MI (538.1) and 2.7 times alfa MI (276.9). The omicron S-RBD demonstrated the most vital transmissible strength. The 14 currently proposed medical treatment compounds did not show as the inhibitors to block the omicron S-RBD and ACE-2 binding; instead, they adsorbed at the ACE-2 active site and may inhibit the ACE-2 activity. A modified candidate (Gallo catechin gallate) whose two phenolic hydroxy groups were replaced with two carboxy groups was repulsed from ACE-2, indicating that further modification of medical treatment candidates may produce an effective docking inhibitor.
查看更多>>摘要：As an important kind of environmental endocrine disruptors, 17 beta-Estradiol (E2) plays a major role in affecting the growth of human including sexual characters, pregnancy system, etc. In the modern society, with the threat of abuse in breeding, it is imperative to design sensitive methods for detecting low concentration of E2 in environment. In this work, we constructed a highly sensitive and simple fluorescent aptasenor for detecting E2 via amplification of hybridization chain reaction (HCR) and horseradish peroxidase (HRP). Through the competitions between complementary strand (cmDNA) and E2 to E2 aptamer modified on magnetic beads, the unbound cmDNA would be collected and captured by polystyrene microspheres to induce HCR which brought abundant biotin sites. Subsequently, benefit from the excellent catalytic performance of streptavidin-horseradish peroxidase (SA-HRP), the highly sensitive fluorescence signals could be obtained in low concentration of E2. Under the optimal conditions, the prospered method for E2 detection was shown a good liner range from 1 to 100 pg/mL, with the lower detecting limit of 0.2 pg/mL compared with previous work. In addition, the recovery rates tested in the real samples of milk and water were 99.20%-108.06% and 91.07%-106.13%. In all, the assay may provide a perspective way for highly sensitive detection for various contaminants in the real samples.
查看更多>>摘要：Since novel nutrient sources with high protein content, such as yeast, fungi, bacteria, algae, and insects, are increasingly introduced in the consumer market, safety evaluation studies on their potentially allergenic proteins are required. A pipeline for in silico establishing the sequence-based homology between proteins of spirulina (Arthrospira platensis) and chlorella (Chlorella vulgaris) micro-algae and those included in the AllergenOnline (AO) database (AllergenOnline.org) is described. The extracted proteins were first identified through tryptic peptides analysis by reversed-phase liquid chromatography and high resolution/accuracy Fourier-transform tandem mass spectrometry (RPLC-ESI-FTMS/MS), followed by a quest on the UniProt database. The AO database was subsequently interrogated to assess sequence similarity between identified microalgal proteins and known allergens, based on criteria established by the World Health Organization (WHO) and Food and Agriculture Organization (FAO). A direct search for microalgal proteins already included in allergen databases was also performed using the Allergome database. Six proteins exhibiting a significant homology with food allergens were identified in spirulina extracts. Five of them, i.e., two thioredoxins (D4ZSU6, K1VP15), a superoxide dismutase (C3V3P3), a glyceraldehyde-3-phosphate dehydrogenase (K1W168), and a triosephosphate isomerase (D5A635), resulted from the search on AO. The sixth protein, C-phycocyanin beta subunit (P72508), was directly obtained after examining the Allergome database. Two proteins exhibiting significant sequence homology with food allergens were retrieved in chlorella extracts, viz. calmodulin (A0A2P6TFR8), which is related to troponin c (D7F1Q2), and fructose-bisphosphate aldolase (A0A2P6TDD0). Specific serum screenings based on immunochemical tests should be undertaken to confirm or rule out the allergenicity of the identified proteins.
查看更多>>摘要：The foot-and-mouth disease (FMD) is the most important transboundary viral disease of livestock in the international context, because of its extreme contagiousness, widespread diffusion, and severe impact on animal trade and animal productions. The rapid and on-field detection of the virus responsible for the FMD represents an urgent demand to efficiently control the diffusion of the infection, especially in low resource setting where the FMD is endemic. Colorimetric lateral flow immunoassay (LFIA) is largely used for the development of rapid tests, due to the extreme simplicity, cost-effectiveness, and on-field operation. In this work, two multiplex LFIA devices were designed for the diagnosis of FMD and the simultaneous identification of major circulating serotypes of the FMD virus. The LFIAs relied on the sandwich-type immunoassay and combined a set of well-characterised monoclonal antibodies (mAb) pairs. One LFIA aimed at detecting and identifying O, A and Asia-1 serotypes, the second device enabled the detection and differentiation of the SAT 1 and SAT 2 serotypes. Both devices also incorporated a broad-specific test line reporting on infection from FMDV, regardless the strain and the serotype involved. Accordingly, five and four reactive zones were arranged in the two devices to achieve a total of six simultaneous analyses. The development of the two multiplex systems highlighted for the first time the relevance of the mAb positioning along the LFIA strip in connection with the use of the same or different mAb as capture and detector ligands. In fact, the excess of detector mAb typically employed for increasing the sensitivity of sandwich immunoassay induced a new type of hook effect when combined with the same ligand used as the capture. This effect strongly impacted assay sensitivity, which could be improved by an intelligent alignment of the mAb pairs along the LFIA strip. The analytical and diagnostic performances of the two LFIAs were studied by testing reference FMDV strains grown in cell cultures and some representative field samples (epithelium homogenates). Almost equivalent sensitivity and specificity to those of a reference Ag-ELISA kit were shown, except for the serotype SAT 2. These simple devices are suitable in endemic regions for in-field diagnosis of FMD accompanied by virus serotyping and, moreover, could be deployed and used for rapid confirmation of secondary outbreaks after FMD incursions in free-areas, thus contributing to promptly implement control measures.
查看更多>>摘要：In this work, the effect of sample matrix on electromembrane extraction (EME) was investigated for the first time using cathinones (log P < 1.0) as polar basic model analytes. Ten supported liquid membranes (SLMs) were tested for EME from spiked buffer solutions, urine, and whole blood samples, respectively. For buffer solutions, SLMs containing aromatic solvents provided higher EME recovery than non-aromatic solvents, which confirmed the significance of cation-pi interactions for EME of basic substances. Interestingly, when applied to urine and whole blood samples, aromatic SLMs were less efficient, while non-aromatic SLMs containing abundant hydrogen-bond acidity/basicity were efficient. These observations were explained by SLM fouling, and the antifouling property of the SLM was clearly dependent on the nature of the SLM solvent. Accordingly, a binary SLM containing aromatic 1-ethyl-2-nitrobenzene (ENB) and non-aromatic 1-undecanol (1:1 v/v) was developed. This binary SLM was not prone to fouling, and provided high recoveries of cathinones from urine and whole blood. EME based on this SLM was optimized and evaluated in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS), and the linear ranges with R-2 >= 0.9903 for cathinones in whole blood and urine were 5-200 ng/mL and 1-200 ng/mL, respectively. The LOD and LOQ of cathinones were ranged from 0.12 to 0.54 ng/mL and 0.38-1.78 ng/mL, respectively. The repeatability and accuracy bias at three levels were <= 11% and within 10%, respectively. In addition, the matrix effect ranged from 88% to 118% was also in compliance with guidelines for bioanalytical method validation provided by the European Medicines Agency.
查看更多>>摘要：A self-powered photoelectrochemical (PEC) aptasensor was constructed based on MIL-68(In) derived indium oxide hollow nanotubes (In2O3 HNs) and Ag-doped ZnIn2S4 quantum dots (QDs) as sensing matrix for the ultrasensitive detection of oxytetracycline (OTC). The hollow tube structure of the designed photoelectric active platform provided abundant active sites and a larger specific surface area for the immobilization of target recognition unit. The coupling of Ag:ZnIn2S4 QDs and In2O3 HNs can accelerate the transmit and separation of photoinduced charge, and thus greatly increasing the intensity of photocurrent signal. Then, the well-constructed OTC-aptamer was anchored on the modified photoelectrode as an accurate capturing element, achieving the specific detection of analyte. Under optimal conditions, the photocurrent intensity of the PEC aptasensor decreases linearly, with a linear response range of 10-4 -10 nmol/L, and a limit of detection (LOD) of 3.3 x 10-5 nmol/L (S/N = 3). The developed self-powered aptasensor with excellent reproducibility, stability, and selectivity, provides a potential way to detect antibiotic residues in environmental media.
NSTL
Elsevier