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One‐dimensional zinc oxide nanomaterials (1DZnO) have emerged as promising, cost‐effective nanoplatforms with adjustable properties suitable for electrochemical and optical biosensing applications. In this work, modifications in the inherent photoluminescent response of 1DZnO are harnessed to develop a novel immunosensor tailored for detecting enteropathogenic Escherichia coli. This nanobiosensor demonstrates a modulation in photoluminescence signal, effectively responsive to analyte concentrations ranging from 1 × 102 to 1 × 108 CFU mL−1, with direct visualization of targeted bacterial cells over 1DZnO structures through scanning electron microscopy. The conceptualization of this nanobiosensor is focused on a real‐time contact strategy that can significantly reduce processing and response times for pathogen detection, prospected for emergency scenarios. With this aim, the detection process unfolds in real time, with a mere 5–10 s interaction time, corroborated by the standard polymerase chain reaction approach. This synergistic validation underscores the reliability and precision of the developed biosensor. Notably, the utility of 1DZnO nanoplatforms extends beyond the realm of enteropathogenic E. coli, as the biosensing performance exhibited here holds promise for analogous applications involving other medically pertinent pathogens. This study paves the way for the broader implementation of 1DZnO‐based biosensors in medical diagnostics, offering rapid, sensitive, and real‐time detection capabilities. [ABSTRACT FROM AUTHOR]

The detection of biomarkers with both high sensitivity and specificity is crucial for the diagnosis and treatment of related diseases. However, many current detections employ ex-situ detection method and non-confined condition, thus have many problems, which may eventually lead to inaccurate detection results. Compared to detection in non-confined space, detection in confined space can better reflect the real in-vivo situation. Therefore, the construction of detection for target molecules in confined space has great significance for both theoretical research and practical application. To realize the detection of target molecules in confined space, the probes should accurately enter the confined space where the target molecules reside and interact with the interface. Thus, how to explore and utilize the properties of the interface (for example, bioinspired superwettability) has always been a hot and difficult topic in this field. Herein, the recent advances and our efforts in recent 10 years on detection of bio-target molecules in confined space with superwettable interface have been introduced from the perspective of the detection methods. The suitable and most widely employed detection methods for target molecules in confined spaces are introduced firstly. Then, recent progresses for related detections based on visual, optical, and electrochemical detection methods are presented successively. Finally, the perspective for detection in confined space is discussed for the future development of biochemical detection. [ABSTRACT FROM AUTHOR]

Longitudinal giant magnetoimpedance effect of [Fe21Ni79/Cu]5/Cu/[Fe21Ni79/Cu]5 film element was investigated depends on stray magnetic field of epoxy magnetic composite with 30% weight concentration of iron oxide magnetic microparticles. Configuration of an experiment was a model of thrombus detection in a blood vessel. Stray magnetic field was varied by movement of a magnetic composite above the element perpendicular to the long side. Composite was either magnetized or not to the state of remanence. As the magnetic composite approaches the GMI element, MI ratio curves are smoothed and shifted along the field axis and maximum value of the MI ratio decreases. Magnetic properties of magnetic composite and film element were investigated as well. [ABSTRACT FROM AUTHOR]

After the first visible photoluminescence (PL) from porous silicon (pSi), continuous efforts are made to fabricate Si-based compound nanomaterials embedded in matrices such as oxide, nitride, and carbide to improve optical performance and industrial acceptability. These nanomaterials' functional and desired properties (nanoparticles and quantum dots embedded in matrices) can vary significantly when embedded in technologically relevant matrices. However, exploring the exact emission mechanisms is one of the remaining challenges from the past few decades. To cover this gap, this review discusses the morphological and optoelectronic properties of Si-based compound nanomaterials and their correlation with the quantum confinement effect and different surface states to find precise emission mechanisms. One of the biggest challenges of using silicon nanomaterials in the biological sector is the development of sensitive materials of low/acceptable toxicity for identifying target analytes either inside/outside the biological platforms. In this scenario, silicon-based compound matrices can offer different characteristics and advantages depending on their size configurations and PL emission mechanisms. On the other hand, a proper understanding of these multifaceted silicon nanomaterials' optical properties (emission mechanisms) can be exploited for pathogen detection and in situ applications in cells and tissues, embarking on a new era of bioimaging technology. [ABSTRACT FROM AUTHOR]

Terahertz (THz) waves, which fall between microwaves and infrared bands, possess intriguing electromagnetic properties of non-ionizing radiation, low photon energy, being highly sensitive to weak resonances, and non-polar material penetrability. Therefore, THz waves are extremely suitable for sensing and detecting chemical, pharmaceutical, and biological molecules. However, the relatively long wavelength of THz waves (30~3000 μm) compared to the size of analytes (1~100 nm for biomolecules, <10 μm for microorganisms) constrains the development of THz-based sensors. To circumvent this problem, metasurface technology, by engineering subwavelength periodic resonators, has gained a great deal of attention to enhance the resonance response of THz waves. Those metasurface-based THz sensors exhibit high sensitivity for label-free sensing, making them appealing for a variety of applications in security, medical applications, and detection. The performance of metasurface-based THz sensors is controlled by geometric structure and material parameters. The operating mechanism is divided into two main categories, passive and active. To have a profound understanding of these metasurface-assisted THz sensing technologies, we review and categorize those THz sensors, based on their operating mechanisms, including resonators for frequency shift sensing, nanogaps for enhanced field confinement, chirality for handedness detection, and active elements (such as graphene and MEMS) for advanced tunable sensing. This comprehensive review can serve as a guideline for future metasurfaces design to assist THz sensing and detection. [ABSTRACT FROM AUTHOR]

The first example of inkjet‐printed, electrolyte‐gated organic field‐effect transistors, fabricated on flexible polyimide substrates is presented. The interdigitated source and drain electrodes, and the coplanar gate electrodes, are inkjet‐printed using a homemade gold nanoparticle ink. A semiconducting ink based on the p‐type, organic semiconductor poly[2,5‐(2‐octyldodecyl)‐3,6‐diketopyrrolopyrrole‐alt‐5,5‐(2,5‐di(thien‐2‐yl)thieno [3,2‐b] thiophene)] (DPP‐DTT) is formulated and inkjet‐printed onto the channel. The performances of inkjet‐printed, coplanar devices are compared to those of transistors whose gate electrode consists in a metallic wire inserted in the electrolyte. Printed transistors show excellent electrical properties with field‐effect mobility as high as 0.04 cm2 V−1 s−1. The electrical behavior of inkjet‐printed, coplanar devices is also modeled using the Nernst–Planck–Poisson (NPP) equations, where the output and transfer curves are calculated based on the charge and potential distribution inside the device. Good quantitative agreement between the simulation and experiments is achieved, outlining the attainable use of NPP simulations as predictive tools for device design and optimization. To demonstrate an example of application, printed transistors are functionalized for the detection of complementary DNA strands. This study opens an avenue for the next generation of low‐cost, flexible sensors and circuits, both through experimental studies and device modeling. [ABSTRACT FROM AUTHOR]

Advances in nanotechnology and medical science have spurred the development of engineered nanomaterials and nanoparticles with particular focus on their applications in biomedicine. In particular, gold nanoparticles (AuNPs) have been the focus of great interest, due to their exquisite intrinsic properties, such as ease of synthesis and surface functionalization, tunable size and shape, lack of acute toxicity and favorable optical, electronic, and physicochemical features, which possess great value for application in biodetection and diagnostics purposes, including molecular sensing, photoimaging, and application under the form of portable and simple biosensors (e.g., lateral flow immunoassays that have been extensively exploited during the current COVID‐19 pandemic). We shall discuss the main properties of AuNPs, their synthesis and conjugation to biorecognition moieties, and the current trends in sensing and detection in biomedicine and diagnostics. This article is categorized under:Diagnostic Tools > BiosensingDiagnostic Tools > In Vitro Nanoparticle‐Based SensingDiagnostic Tools > In Vivo Nanodiagnostics and Imaging [ABSTRACT FROM AUTHOR]

An optimized, well-tested and validated targeted genomic sequencing-based high-throughput assay is currently not available ready for routine biodefense and biosurveillance applications. Earlier, we addressed this gap by developing and establishing baseline comparisons of a multiplex end-point Polymerase Chain Reaction (PCR) assay followed by Oxford Nanopore Technology (ONT) based amplicon sequencing to real time PCR and customized data processing. Here, we expand upon this effort by identifying the optimal ONT library preparation method for integration into a novel software platform ONT-DART (ONT-Detection of Amplicons in Real-Time). ONT-DART is a dockerized, real-time, amplicon-sequence analysis workflow that is used to reproducibly process and filter read data to support actionable amplicon detection calls based on alignment metrics, within sample statistics, and no-template control data. This analysis pipeline was used to compare four ONT library preparation protocols using R9 and Flongle (FL) flow cells. The two 4-Primer methods tested required the shortest preparation times (5.5 and 6.5 h) for 48 libraries but provided lower fidelity data. The Native Barcoding and Ligation methods required longer preparation times of 8 and 12 h, respectively, and resulted in higher overall data quality. On average, data derived from R9 flow cells produced true positive calls for target organisms more than twice as fast as the lower throughput FL flow cells. These results suggest that utilizing the R9 flowcell with an ONT Native Barcoding amplicon library method in combination with ONT-DART platform analytics provides the best sequencing-based alternative to current PCR-based biodetection methods. [ABSTRACT FROM AUTHOR]

Microorganisms are omnipresent and inseparable from our life. Many of them are beneficial to humans, while some are not. Importantly, foods and beverages are susceptible to microbial contamination, with their toxins causing illnesses and even death in some cases. Therefore, monitoring and detecting harmful microorganisms are critical to ensuring human health and safety. For several decades, many methods have been developed to detect and monitor microorganisms and their toxicants. Conventionally, nucleic acid analysis and antibody-based analysis were used to detect pathogens. Additionally, diverse chromatographic methods were employed to detect toxins based on their chemical and structural properties. However, conventional techniques have several disadvantages concerning analysis time, sensitivity, and expense. With the advances in biotechnology, new approaches to detect pathogens and toxins have been reported to compensate for the disadvantages of conventional analysis from different research fields, including electrochemistry, nanotechnology, and molecular biology. Among them, we focused on the recent studies of transcription factor (TF)-based biosensors to detect microorganisms and discuss their perspectives and applications. Additionally, the other biosensors for detecting microorganisms reported in recent studies were also introduced in this review. [ABSTRACT FROM AUTHOR]

The dependence of a longitudinal giant magnetoimpedance (GMI) effect of a [Fe21Ni79/Cu]5/Cu/[Fe21Ni79/Cu]5 film element on the position of a magnetic insert is investigated in a model experiment on detecting a clot in a blood vessel. The magnetic insert is an epoxy composite containing 30 wt.% of the iron oxide microparticles. An investigation of the magnetic properties of the film element and the magnetic composite cylinder-shaped sample is performed. The magnetic insert is placed at a distance of 1.1 mm above the film surface and can be displaced perpendicular to its longer side with a step of 1 mm. As soon as the magnetic insert approaches the element, the maximum value of magnetoimpedance (MI) ratio for resistance is observed to decrease, and there is a shift of the curves of the MI ratio for resistance along the external magnetic field lines. The magnetic stray fields influencing the film in each position of the magnetic insert are simulated with the Comsol software and compared with the GMI response. [ABSTRACT FROM AUTHOR]

This study reports the synthesis and characterization of the red nanophosphors Zn2SiO4:Eu3+ (ZSO:Eu3+) and Mg2TiO4:Mn4+ (MTO:Mn4+). The use of phosphors as a fluorescence label for lateral flow immunochromatographic assay (LFIA) has also been described. The optimal photoluminescence (PL) for ZSO:Eu3+ was obtained when it was synthesized with 7 mol% of Eu3+ and annealed at 1100 °C for 1 h. Long fluorescence lifetime (1.01 ms), high activation energy E a (0.28 eV), and low PL degeneration (10% at 110 °C) are the characteristics of ZSO:Eu3+. MTO:Mn4+ also exhibited high PL intensity along with a high Ea of 0.32 eV. The emission wavelengths of phosphors are biocompatible with the optical bio-window of tissues. When human immunoglobulin G (human IgG) at a constant concentration of 100 μg/mL was used for detection, the PL ratios of the test line to the control line were 2.15 and 2.28 for the ZSO:Eu3+- and MTO:Mn4+-labeled LFIA, respectively. Thus, the ZSO:Eu3+ and MTO:Mn4+ nanophosphors are capable of human IgG recognition and are the promising candidates as fluorescent labels for on-site rapid optical biodetection. [ABSTRACT FROM AUTHOR]

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Dioxins are the most toxic known environmental pollutants and are mainly formed by human activities. Due to their structural stability, dioxins persist for extended periods and can be transported over long distances from their emission sources. Thus, dioxins can be accumulated to considerable levels in both human and animal food chains. Along with sediments, soils are considered the most important reservoirs of dioxins. Soil microorganisms are therefore highly exposed to dioxins, leading to a range of biological responses that can impact the diversity, genetics and functional of such microbial communities. Dioxins are very hydrophobic with a high affinity to lipidic macromolecules in exposed organisms, including microbes. This review summarizes the genetic, molecular and biochemical impacts of dioxins on the lipid metabolism of soil microbial communities and especially examines modifications in the composition and architecture of cell membranes. This will provide a useful scientific benchmark for future attempts at soil ecological risk assessment, as well as in identifying potential dioxin-specific-responsive lipid biomarkers. Finally, potential uses of lipid-sequestering microorganisms as a part of biotechnological approaches to the bio-management of environmental contamination with dioxins are discussed. [ABSTRACT FROM AUTHOR]

Detection of a single base mutation in Mycobacterium tuberculosis DNA can provide fast and highly specific diagnosis of antibiotic-resistant tuberculosis. Mutation-specific ligation of padlock probes (PLPs) on the target followed by rolling circle amplification (RCA) is highly specific, but challenging to integrate in a simple microfluidic device due to the low temperature stability of the phi29 polymerase and the interference of phi29 with the PLP annealing and ligation. Here, we utilized the higher operation temperature and temperature stability of Equiphi29 polymerase to simplify the integration of the PLP ligation and RCA steps of an RCA assay in two different strategies performed at uniform temperature. In strategy I, PLP annealing took place off-chip and the PLP ligation and RCA were performed in one pot and the two reactions were clocked by a change of the temperature. For a total assay time of about 1.5 h, we obtained a limit of detection of 2 pM. In strategy II, the DNA ligation mixture and the RCA mixture were separated into two chambers on a microfluidic disc. After on-disc PLP annealing and ligation, the disc was spun to mix reagents and initiate RCA. For a total assay time of about 2 h, we obtained a limit of detection of 5 pM. [ABSTRACT FROM AUTHOR]

Nanotechnology offers the possibility of operating on the same scale length at which biological processes occur, allowing to interfere, manipulate or study cellular events in disease or healthy conditions. The development of hybrid nanostructured materials with a high degree of chemical control and complex engineered surface including biological targeting moieties, allows to specifically bind to a single type of molecule for specific detection, signaling or inactivation processes. Magnetite nanostructures with designed composition and properties are the ones that gather most of the designs as theragnostic agents for their versatility, biocompatibility, facile production and good magnetic performance for remote in vitro and in vivo for biomedical applications. Their superparamagnetic behavior below a critical size of 30 nm has allowed the development of magnetic resonance imaging contrast agents or magnetic hyperthermia nanoprobes approved for clinical uses, establishing an inflection point in the field of magnetite based theragnostic agents. [ABSTRACT FROM AUTHOR]

Background: The state-of-the-art in nucleic acid based biodetection continues to be polymerase chain reaction (PCR), and many real-time PCR assays targeting biodefense pathogens for biosurveillance are in widespread use. These assays are predominantly singleplex; i.e. one assay tests for the presence of one target, found in a single organism, one sample at a time. Due to the intrinsic limitations of such tests, there exists a critical need for high-throughput multiplex assays to reduce the time and cost incurred when screening multiple targets, in multiple pathogens, and in multiple samples. Such assays allow users to make an actionable call while maximizing the utility of the small volumes of test samples. Unfortunately, current multiplex real-time PCR assays are limited in the number of targets that can be probed simultaneously due to the availability of fluorescence channels in real-time PCR instruments. Results: To address this gap, we developed a pipeline in which the amplicons produced by a 14-plex end-point PCR assay using spiked samples were subsequently sequenced using Nanopore technology. We used bar codes to sequence multiple samples simultaneously, leading to the generation and subsequent analysis of sequence data resulting from a short sequencing run time (< 10 min). We compared the limits of detection (LoD) of real-time PCR assays to Oxford Nanopore Technologies (ONT)-based amplicon sequencing and estimated the sample-to-answer time needed for this approach. Overall, LoDs determined from the first 10 min of sequencing data were at least one to two orders of magnitude lower than real-time PCR. Given enough time, the amplicon sequencing approach is approximately 100 times more sensitive than real-time PCR, with detection of amplicon specific reads even at the lowest tested spiking concentration (around 2.5–50 Colony Forming Units (CFU)/ml). Conclusions: Based on these results, we propose amplicon sequencing assay as a viable alternative to replace the current real-time PCR based singleplex assays for higher throughput biodefense applications. We note, however, that targeted amplicon specific reads were not detectable even at the highest tested spike concentrations (2.5 X 104–5.0 X105 CFU/ml) without an initial amplification step, indicating that PCR is still necessary when utilizing this protocol. [ABSTRACT FROM AUTHOR]

Quantum dots (QD) are being evaluated as inorganic nanoparticles for both in vitro and in vivo optical imaging. They are also used as sensors or vehicles for targeted drug delivery combined with optical imaging. In this study, we demonstrated that glutathione‐coated Ag2S QDs (GSH‐Ag2S QDs) act as a substrate analogue of glutathione S‐transferase (GST) enzymes for the first time in the literature. The GSTs belong to a major group of detoxification enzymes involved in the detoxification metabolism responsible for the protection of cells against reactive oxygen species (ROS) or electrophiles. GST isozymes are impaired in the various diseases such as neurological diseases and cancer. We evaluated the interaction of GST‐pi enzyme with GSH‐Ag2S QDs, which have never been studied in the literature before, using both fluorometric and spectrophotometric methods. Our data showed that GSH‐Ag2S QDs gave reaction with GST enzyme as a substrate analogue. In conclusion, our data may help to guide researchers for further development of sensing systems for GST activity which is impaired in various diseases including cancer. [ABSTRACT FROM AUTHOR]

A colorimetric assay is described for simultaneous detection of multiple analytes related to food safety. It is based on the use of a sandwich aptasensor and terminal deoxynucleotidyl transferase (TdT) which produces a primer for subsequent rolling circle amplification (RCA). Two split aptamer fragments (Apt1 and Apt2) are firstly immobilized, Apt1 on gold nanoparticles (AuNPs), and Apt2 on magnetic beads (MBs). They are then used in a sandwich aptasensor. In the presence of analyte, two probes could specifically recognize target and form a ternary assembly, and the magnetic beads also act to separate rapidly and enrich the target. Then, the extension of template-free DNA is triggered by TdT at the exposed 3′-hydroxy terminals of Apt1. This produces polyA sequences that serve as primers for subsequent RCA. The product of RCA is hybridized with a complementary horse radish peroxidase (HRP) DNA probe. HRP catalyzes the H2O2-mediated oxidation of tetramethylbenzidine (TMB) and forms a blue chromogenic product. After magnetic separation, the absorption values of the blue product in the supernatant are measured at a wavelength of 600 nm. Based on this dual amplification mechanism, the assay was applied to multiplexed determination of enrofloxacin (ENR), lead(II), Escherichia coli O157:H7 and tropomyosin. Exemplarily, ENR is detectable at concentrations down to 2.5 pg mL−1 with a linear range that extends from 1 pg mL−1 to 1 μg·mL−1. The assay was validated by analysis of spiked fish samples. Recoveries range between 87.5 and 92.1%. [ABSTRACT FROM AUTHOR]

Although a variety of whole-cell biosensors and biosorbents have been developed for detection and removal of heavy metal contaminants, few whole cells can be applied to both monitoring and remediation of copper pollution in water. In this study, a modified plasmid was constructed by incorporating a copper-sensing element and a copper-adsorbing element into a temperature-inducible plasmid, pBV220. This plasmid was subsequently transformed into an engineered Escherichia coli strain lacking copA and cueO. This dual-functional E. coli cell selectively responded to copper ions with a linear detection range of 0.01–25 μM at 37 °C and could express surface-displayed CueR when treated at 42 °C without any costly chemical inducers. The display of CueR on the cell surface specifically enhanced its copper adsorption capacity and rapidly removed copper ions from aqueous solutions. In addition, the CueR surface-displayed cells could be regenerated by adsorption-desorption cycles via pH regulation. Moreover, by simply using two different temperatures, the detection or adsorption of copper using this dual-functional whole cell was achieved without any cross-interference. Most importantly, it provided highly sensitive, accurate quantification, and effective removal of copper in real environmental water samples. Thus, this E. coli cell can be used for large-scale detection and remediation of copper pollutants. [ABSTRACT FROM AUTHOR]

Self-assembled three-dimensional (3-D) colloidal crystals have been extensively utilized in nanolithography for fabricating a wide spectrum of functional periodic nanostructures for important technological applications. Here a simple and versatile way is reported using silica colloids to prepare close-packed particle multilayers by the Langmuir– Blodgett technique to create highly ordered 3-D colloidal crystals with unusual nanoarrays on a large variety of substrates. Successive deposition of monolayers using amphiphilic molecules in this approach allows control of the layer thickness, particle size, and especially packing structure of them in each layer. It has been demonstrated that the Langmuir–Blodgett film’s transparency and high antiglare optical properties are preserved even after multiple colloidal particle transfers on both straight and curved surfaces. The surface prepared by the Langmuir–Blodgett technique shows remarkable sensitivity for potential biodetection applications. These colloidal templated films have an optical response time of less than 40 s against essential bioproduct vapors. It is shown that the results are consistent with the Kelvin equation (ln P/Po = –2 Vl γ/rRT) when the liquid water layer is getting thicker. [ABSTRACT FROM AUTHOR]

Introduction: An update on the uses and applications of the non-cross-linking (NCL) hybridization assay based on the spectral modulation of gold nanoparticles (AuNPs) are presented, emphasizing DNA and RNA detection. Areas covered: Nanotechnology is strongly impacting the way we address diagnostics and therapeutics. In fact, nanoscale devices and particles have been used in a variety of platforms for improved biosensing and, more interestingly, for molecular diagnostics. AuNPs have been used in a great diversity of DNA and RNA detection strategies that are based on their nanoscale properties. Their unique optical properties have put them at the forefront of colorimetric sensing platforms. Among these, those relying on the NCL mechanism using DNA-modified AuNPs have shown remarkable versatility and simplicity for molecular detection of human pathogens, identification of single base alterations at the basis of human disease, gene expression, among others. Application of the NCL assay to molecular diagnostics will be discussed considering the challenges for validation and clinically relevant targets. Expert commentary: Integration of the NCL approach using AuNPs into chip biosensing platforms, projecting miniaturization and portability, will be addressed in terms of the future, i.e. clinical validation and translation to market. [ABSTRACT FROM AUTHOR]

Abstract: Significant progress on upconversion‐nanoparticle (UCNP)‐based probes is witnessed in recent years. Compared with traditional fluorescent probes (e.g., organic dyes, metal complexes, or inorganic quantum dots), UCNPs have many advantages such as non‐autofluorescence, high chemical stability, large light‐penetration depth, long lifetime, and less damage to samples. This article focuses on recent achievements in the usage of lanthanide‐doped UCNPs as efficient probes for biodetection since 2014. The mechanisms of upconversion as well as the luminescence resonance energy transfer process is introduced first, followed by a detailed summary on the recent researches of UCNP‐based biodetections including the detection of inorganic ions, gas molecules, reactive oxygen species, and thiols and hydrogen sulfide. [ABSTRACT FROM AUTHOR]

Bacterial cell immobilization is a novel technique used in many areas of biosciences and biotechnology. Iron oxide nanoparticles have attracted much attention in bacterial cell immobilization due to their unique properties such as superparamagnetism, large surface area to volume ratio, biocompatibility and easy separation methodology. Adhesion is the basis behind many immobilization techniques and various types of interactions determine bacterial adhesion. Efficiency of bacterial cell immobilization using iron oxide nanoparticles (IONs) generally depends on the physicochemical properties of the IONs and surface properties of bacterial cells as well as environmental/culture conditions. Bacteria exhibit various metabolic responses upon interaction with IONs, and the potential applications of iron oxide nanoparticles in bacterial cell immobilization will be discussed in this work. [ABSTRACT FROM AUTHOR]

Protein detection and quantification play critical roles in both basic research and clinical practice. Current detection platforms range from the widely used ELISA to more sophisticated, and more expensive, approaches such as digital ELISA. Despite advances, there remains a need for a method that combines the simplicity and cost-effectiveness of ELISA with the sensitivity and speed of modern approaches in a format suitable for both laboratory and rapid, point-of-care applications. Building on recent developments in DNA structural nanotechnology, we introduce the nanoswitch-linked immunosorbent assay (NLISA), a detection platform based on easily constructed DNA nanodevices that change conformation upon binding to a target protein with the results read out by gel electrophoresis. NLISA is surface-free and includes a kinetic-proofreading step for purification, enabling both enhanced sensitivity and reduced cross-reactivity. We demonstrate femtomolar-level detection of prostate-specific antigen in biological fluids, as well as reduced cross-reactivity between different serotypes of dengue and also between a single-mutation and wildtype protein. NLISA is less expensive, uses less sample volume, is more rapid, and, with no washes, includes fewer hands-on steps than ELISA, while also achieving superior sensitivity. Our approach also has the potential to enable rapid point-of-care assays, as we demonstrate by performing NLISA with an iPad/ iPhone camera for imaging. [ABSTRACT FROM AUTHOR]

The use of in vitro assays for the biodetection of endocrine active substances (EAS) aims to reduce and replace in vivo studies required for regulatory assessment. However, this approach often fails to take into account the role of biotransformation on the activity of the test substances. A method introducing an S9 metabolic system into the CALUX-reporter gene assays for estrogen receptor α- and anti-androgen receptor-mediated activities was developed. Methoxychlor, which exhibits increased estrogenic and anti-androgenic activities after biotransformation, was used to establish the method in ERα and anti-AR CALUX. For the anti-androgenic assay, stanozolol was used as a competing agonist not metabolized by S9. The method was first applied in both agonist and antagonist modes to methoxychlor and bisphenol A, as positive and negative controls, respectively. Then, benzo(a)pyrene and flutamide were also tested for their bioactivation potential. Co-treatment with S9 successfully increased the ERα agonist and AR antagonist potency of methoxychlor; no change was observed for bisphenol A. Incubation with S9 also enhanced the anti-androgenic activity of flutamide. Interestingly, the metabolism of benzo(a)pyrene by S9 resulted in a greatly increased estrogen receptor-mediated transcriptional activation, while the shift in EC50 was only minor. It is likely that both enzyme kinetics and metabolite stability affect the composition of the final metabolite mixture and thus influence the observed effects. Together these results demonstrate the relevance of including biotransformation in in vitro bioassays for the detection of EAS. [ABSTRACT FROM AUTHOR]

Photonic crystal (PC) materials exhibit unique structural colors that originate from their intrinsic photonic band gap. Because of their highly ordered structure and distinct optical characteristics, PC-based biomaterials have advantages in the multiplex detection, biomolecular screening and real-time monitoring of biomolecules. In addition, PCs provide good platforms for drug loading and biomolecule modification, which could be applied to biosensors and biological carriers. A number of methods are now available to fabricate PC materials with variable structure colors, which could be applied in biomedicine. Emphasis is given to the description of various applications of PC materials in biomedicine, including drug delivery, biodetection and tumor screening. We believe that this article will promote greater communication among researchers in the fields of chemistry, material science, biology, medicine and pharmacy. [ABSTRACT FROM AUTHOR]

Portable detection and quantitation methods for Bacillus anthracis (anthrax) spores in pure culture or in environmental samples are lacking. Here, an amperometric immunoassay has been developed utilizing immunomagnetic separation to capture the spores and remove potential interferents from test samples followed by amperometric measurement on a field-portable instrument. Antibody-conjugated magnetic beads and antibody-conjugated glucose oxidase were used in a sandwich format for the capture and detection of target spores. Glucose oxidase activity of spore pellets was measured indirectly via amperometry by applying a bias voltage after incubation with glucose, horseradish peroxidase, and the electron mediator 2,20-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid). Target capture was mediated by polyclonal antisera, whereas monoclonal antibodies were used for signal generation. This strategy maximized sensitivity (500 target spores, 5000 cfu/mL), while also providing a good specificity for Bacillus anthracis spores. Minimal signal deviation occurs in the presence of environmental interferents including soil and modified pH conditions, demonstrating the strengths of immunomagnetic separation. The simultaneous incubation of capture and detection antibodies and rapid substrate development (5 min) result in short sample-to-signal times (less than an hour). With attributes comparable or exceeding that of ELISA and LFDs, amperometry is a low-cost, low-weight, and practical method for detecting anthrax spores in the field. [ABSTRACT FROM AUTHOR]

Non-crosslinking (NCL) approaches using DNA-modified gold nanoparticles for molecular detection constitute powerful tools with potential implications in clinical diagnostics and tailored medicine. From detection of pathogenic agents to identification of specific point mutations associated with health conditions, these methods have shown remarkable versatility and simplicity. Herein, the NCL hybridization assay is broken down to the fundamentals behind its assembly and detection principle. Gold nanoparticle synthesis and derivatization is addressed, emphasizing optimal size homogeneity and conditions for maximum surface coverage, with direct implications in downstream detection. The detection principle is discussed and the advantages and drawbacks of different NCL approaches are discussed. Finally, NCL-based applications for molecular detection of clinically relevant loci and potential integration into more complex biosensing platforms, projecting miniaturization and portability are addressed. [ABSTRACT FROM PUBLISHER]

Recent advances in biosensing technologies present great potential for medical diagnostics, thus improving clinical decisions. However, creating a label-free general sensing platform capable of detecting multiple biotargets in various clinical specimens over a wide dynamic range, without lengthy sample-processing steps, remains a considerable challenge. In practice, these barriers prevent broad applications in clinics and at patients' homes. Here, we demonstrate the nanoplasmonic electrical field-enhanced resonating device (NE2RD), which addresses all these impediments on a single platform. The NE2RD employs an immunodetection assay to capture biotargets, and precisely measures spectral color changes by their wavelength and extinction intensity shifts in nanoparticles without prior sample labeling or preprocessing. We present through multiple examples, a label-free, quantitative, portable, multitarget platform by rapidly detecting various protein biomarkers, drugs, protein allergens, bacteria, eukaryotic cells, and distinct viruses. The linear dynamic range of NE2RD is five orders of magnitude broader than ELISA, with a sensitivity down to 400 fg/mL This range and sensitivity are achieved by self-assembling gold nanoparticles to generate hot spots on a 3D-oriented substrate for ultrasensitive measurements.We demonstrate that this precise platform handles multiple clinical samples such as whole blood, serum, and saliva without sample preprocessing under diverse conditions of temperature, pH, and ionic strength. The NE2RD's broad dynamic range, detection limit, and portability integrated with a disposable fluidic chip have broad applications, potentially enabling the transition toward precision medicine at the pointof- care or primary care settings and at patients' homes. [ABSTRACT FROM AUTHOR]

The s-triazine herbicides are compounds which can disseminate into soils and water. Due to their toxic effects on living organisms, their concentrations in drinking water are legislated by WHO recommendations. Here we have developed for the first time, to the best of our knowledge, an alternative method for physicochemical quantification using two bioluminescent bacterial biosensors: E. coli SM003 for cyanuric acid detection and E. coli SM004 for both atrazine and cyanuric acid detection. The concentration of cyanuric acid detection for E. coli SM003 ranges from 7.83μM to 2.89 mM, and for E. coli SM004 ranges from 0.22 to 15μM. Moreover, atrazine detection by E. coli SM004 ranges from 1.08 to 15μM. According to WHO recommendations, the cyanuric acid detection range is sensitive enough to discriminate between polluted and drinking water. Nevertheless, the detection of atrazine by E. coli SM004 is only applicable for high concentrations of contaminants. [ABSTRACT FROM AUTHOR]

In many cases of envenoming following snake bite, the snake responsible for the accident remains unidentified; this frequently results in difficulty deciding which antivenom to administer to the systemically-envenomed victim, especially when only monospecific antivenoms are available. Normally the specific diagnosis of snake bite can be conveniently made using clinical and laboratory methods. Where clinical diagnosis depends upon the recognition of specific signs of envenoming in the patient, laboratory diagnosis is based on the changes which occur in envenomed victims including the detection of abnormalities in blood parameters, presence/absence of myoglobinuria, changes in certain enzyme levels, presence/absence of neurotoxic signs and the detection in the blood of specific venom antigens using immunologically-based techniques, such as enzyme immunoassay. It is the latter which is the main subject of this review, together with the application of techniques currently used to objectively assess the effectiveness of new and existing antivenoms, to assess first aid measures, to investigate the possible use of such methods in epidemiological studies, and to detect individual venom components. With this in mind, we have discussed in some detail how such techniques were developed and how they have helped in the treatment of envenoming particularly and in venom research in general. [ABSTRACT FROM AUTHOR]

Lanthanide-doped upconversion nanoparticles (UCNPs) have shown great promise in bioapplications. Exploring new host materials to realize efficient upconversion luminescence (UCL) output is a goal of general concern. Herein, we develop a unique strategy for the synthesis of novel LiLuF4:Ln3+ core/shell UCNPs with typically high absolute upconversion quantum yields of 5.0 % and 7.6 % for Er3+ and Tm3+, respectively. Based on our customized UCL biodetection system, we demonstrate for the first time the application of LiLuF4:Ln3+ core/shell UCNPs as sensitive UCL bioprobes for the detection of an important disease marker β subunit of human chorionic gonadotropin (β-hCG) with a detection limit of 3.8 ng mL−1, which is comparable to the β-hCG level in the serum of normal humans. Furthermore, we use these UCNPs in proof-of-concept computed tomography imaging and UCL imaging of cancer cells, thus revealing the great potential of LiLuF4:Ln3+ UCNPs as efficient nano-bioprobes in disease diagnosis. [ABSTRACT FROM AUTHOR]

Aims To evaluate the sensitivity and specificity of the BioFire Diagnostics FilmArray® system in combination with their Biothreat Panel for the detection of Bacillus anthracis ( Ba), Francisella tularensis ( Ft) and Yersinia pestis ( Yp) DNA, and demonstrate the detection of Ba spores. Methods and Results DNA samples from Ba, Ft and Yp strains and near-neighbours, and live Ba spores were analysed using the FilmArray® Biothreat Panel, a multiplexed PCR-based assay for 17 pathogens and toxins. Sensitivity studies with DNA indicate that the limit of detection is 250 genome equivalents ( GEs) per sample or lower. Furthermore, the identification of Ft, Yp or Bacillus species was made in 63 of 72 samples tested at 25 GE or less. With samples containing 25 CFU of Ba Sterne spores, at least one of the two possible Ba markers was identified in all samples tested. We observed no cross-reactivity with near-neighbour DNAs. Conclusions Our results indicate that the FilmArray® Biothreat Panel is a sensitive and selective assay for detecting the genetic signatures of Ba, Ft and Yp. Significance and Impact of the Study The FilmArray® platform is a complete sample-to-answer system, combining sample preparation, PCR and data analysis. This system is particularly suited for biothreat testing where samples need to be analysed for multiple biothreats by operators with limited training. [ABSTRACT FROM AUTHOR]

Copyright of Canadian Journal of Plant Pathology is the property of Taylor & Francis Ltd and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Quantitative polymerase chain reaction (qPCR) assays used in pathogen detection require rigorous methods development including characterizing DNA extraction products. A DNA extract characterization process is demonstrated using DNA extracted from five different cells types (two Gram-negatives: Escherichia coli, and Burkholderia thailandensis, spores and vegetative cells from the Gram-positive Bacillus cereus, and yeast Saccharomyces cerevisiae) with six different methods. Results: DNA extract quantity (concentration and extraction efficiency) and quality (purity and intactness) varied by cell type and extraction method enabling the demonstration of different DNA characterization methods. DNA purity was measured using UV spectroscopy, where the A260/A280 and A260/A230 ratios are indicators of different contaminants. Reproducibility of UV spectroscopy measurements decreased for DNA concentrations less than 17.5 ng/µL. Forty-seven extracts had concentrations greater than 17.5 ng/µL, 25 had A260/A280 above 2.0, and 28 had A260/A230 ratios below 1.8 indicating RNA and polysaccharide contamination respectively. Based on a qPCR inhibition assay the contaminants did not inhibit PCR. Extract intactness was evaluated using microfluidic gel electrophoresis. Thirty-five samples had concentrations above the limit of quantification (LOQ, roughly 11 ng/ µL), 93.5% of the DNA was larger than 1kb and 1% was smaller than 300 bp. Extract concentrations ranged from 1502.2 ng/µL to below the LOQ when UV spectroscopy, fluorometry, and qPCR were used. LOQ for UV spectroscopic and fluorometric measurements were 3.5 ng/µL and 0.25 ng/µL respectively. The qPCR LOQ varied by cell type (5.72 × 10-3 ng/µL for E. coli, 2.66×10-3 ng/µL, for B. cereus, 3.78×10-3 ng/µL for B. thailandensis, and 7.67 × 10-4 ng/µL for S. cerevisiae). A number of samples were below the UV spectroscopy (n = 27), flurometry (n = 15), and qPCR (n = 3) LOQ. Conclusion: The presented DNA extract characterization process provides measures of DNA quantity and quality applicable to microbial detection methods development and validation studies. Evaluating DNA quality and quantity results in a better understanding of process LOD and contributing factors to suboptimal assay performance. The samples used demonstrated the use of different DNA characterization methods presented but did not encompass the full range of DNA extract characteristics. [ABSTRACT FROM AUTHOR]

In this study, a new approach for the preparation of a fluorescent europium(III) complex-doped silica nanoparticles has been developed. The synthesis process involved the following steps: (1) preparing silica nanoparticles by water-in-oil microemulsion method, (2) dyeing the spherical silica particles by europium(III): naphtoyltrifluoroacetone (NTA):trioctylphosphineoxide (TOPO), (3) adsorbing polyvinylpyrrolidone (PVP) onto the core structure and growing silica on PVP surface. The as-prepared nanoparticles exhibited stronger emission intensity, higher photo- and chemical stability. Despite the fact that europium(III) complex was doped into the nanoparticles, its fluorescence properties such as a wide Stokes shift, a narrow emission peak, and long fluorescence lifetime, were retained. The nanoparticles are uniform in shape and size (50 ± 5 nm in diameter). This study could provide new avenue for the fabrication of Eu: NTA:TOPO-based nanoparticles, facilitating their application in bioassay issues. [ABSTRACT FROM AUTHOR]

We explore a new calibration-free approach to biodetection based on whispering gallery modes (WGMs) without a reference measure and relative shifts. Thus, the requirement to keep track of the sensor position is removed, and a freely moving population of fluorophore-doped polystyrene microspheres can now fulfill this role of sensing resonator. Breaking free from fixed surface-based biosensing promotes adhesion between the microsphere sensors and the analytes since both can now be thoroughly mixed. The 70-nm-wide spectrum of green fluorescent microbeads allows us to monitor over 20 WGMs simultaneously without needing evanescent light coupling into the microspheres, hence enabling remote sensing. Since the exact radius of each microsphere is unknown a priori, it requires algorithmic analyses to obtain a reliable result for the refractive index of a solution. We first test our approach with different solutions of alcohol in water obtaining 3 × 10−4 precision on the refractive index at lower concentrations. Then, the solutions of bacterial spores in water yield clear evidence of biodetection in the statistical analysis of WGMs from 50 microspheres. To extend the fluorescence spectral range of our WGM sensors, we present preliminary results on coating microspheres with CdSe/ZnS quantum dots. [ABSTRACT FROM AUTHOR]

Lanthanide-doped nanoparticles possess numerous advantages including tunable luminescence emission, narrow peak width and excellent optical and thermal stability, especially concerning the long lifetime from microseconds to milliseconds. Differing from other shorter-lifetime fluorescent nanomaterials, the long lifetime of lanthanide-doped nanomaterials is independent with background fluorescence interference and biological tissue depth. This review presents the recent advances in approaches to regulating the lifetime and applications of bioimaging and biodetection. We begin with the introduction of the strategies for regulating the lifetime by modulating the core–shell structure, adjusting the concentration of sensitizer and emitter, changing energy transfer channel, establishing a fluorescence resonance energy transfer pathway and changing temperature. We then summarize the applications of these nanoparticles in biosensing, including ion and molecule detecting, DNA and protease detection, cell labeling, organ imaging and thermal and pH sensing. Finally, the prospects and challenges of the lanthanide lifetime regulation for fundamental research and practical applications are also discussed. [ABSTRACT FROM AUTHOR]

Copyright of Umweltwissenschaften und Schadstoff-Forschung is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Oral cancer belongs to the group of head and neck cancers, and, despite its large diffusion, it suffers from low consideration in terms of prevention and early diagnosis. The main objective of the SMILE platform is the development of a low-cost device for oral cancer early screening with features of high sensitivity, specificity, and ease of use, with the aim of reaching a large audience of possible users and realizing real prevention of the disease. To achieve this goal, we realized two microfluidic devices exploiting low-cost materials and processes. They can be used in combination or alone to obtain on-chip sample preparation and/or detection of circulating tumor cells, selected as biomarkers of oral cancer. The realized devices are completely transparent with plug-and-play features, obtained thanks to a highly customized architecture which enables users to easily use them, with potential for a common use among physicians or dentists with minimal training. [ABSTRACT FROM AUTHOR]

The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs and supply shortages. Antigen tests that bind and detect the surface proteins of a virus are rapid and scalable but suffer from high false negative rates. To address this problem, an inexpensive, simple, and robust 60-minute do-it-yourself (DIY) workflow to detect viral RNA from nasal swabs or saliva with high sensitivity (0.1 to 2 viral particles/μL) and specificity (>97% true negative rate) utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed. ALERT (Accessible LAMP-Enabled Rapid Test) incorporates the following features: (1) increased shelf-life and ambient temperature storage, compared to liquid reaction mixes, by using wax layers to isolate enzymes from other reagents; (2) improved specificity compared to other LAMP end-point reporting methods, by using sequence-specific QUASR (quenching of unincorporated amplification signal reporters); (3) increased sensitivity, compared to methods without purification through use of a magnetic wand to enable pipette-free concentration of sample RNA and cell debris removal; (4) quality control with a nasopharyngeal-specific mRNA target; and (5) co-detection of other respiratory viruses, such as influenza B, by multiplexing QUASR-modified RT-LAMP primer sets. The flexible nature of the ALERT workflow allows easy, at-home and point-of-care testing for individuals and higher-throughput processing for labs and hospitals. With minimal effort, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific primer sets can be swapped out for other targets to repurpose ALERT to detect other viruses, microorganisms, or nucleic acid-based markers. [ABSTRACT FROM AUTHOR]

Submicrometer Hall sensors, with Hall cross width of ~250 nm, were fabricated from InAs/AlSb quantum well semiconductor heterostructures. The room-temperature device characteristics were examined by experimental Hall effect and electronic noise measurements combined with analytical calculations. The noise-equivalent magnetic moment resolution of the order of 104 μB/√Hz was obtained at frequencies above ~1 kHz. We show that the devices can achieve single superparamagnetic nanoparticle detection and thus be employed in experiments involving single magnetically labeled biomolecule detection. [ABSTRACT FROM AUTHOR]

The self‐assembly of viral building blocks bears exciting prospects for fabricating new types of bionanoparticles with multivalent protein shells. These enable a spatially controlled immobilization of functionalities at highest surface densities—an increasing demand worldwide for applications from vaccination to tissue engineering, biocatalysis, and sensing. Certain plant viruses hold particular promise because they are sustainably available, biodegradable, nonpathogenic for mammals, and amenable to in vitro self‐organization of virus‐like particles. This offers great opportunities for their redesign into novel "green" carrier systems by spatial and structural synthetic biology approaches, as worked out here for the robust nanotubular tobacco mosaic virus (TMV) as prime example. Natural TMV of 300 x 18 nm is built from more than 2,100 identical coat proteins (CPs) helically arranged around a 6,395 nucleotides ssRNA. In vitro, TMV‐like particles (TLPs) may self‐assemble also from modified CPs and RNAs if the latter contain an Origin of Assembly structure, which initiates a bidirectional encapsidation. By way of tailored RNA, the process can be reprogrammed to yield uncommon shapes such as branched nanoobjects. The nonsymmetric mechanism also proceeds on 3'‐terminally immobilized RNA and can integrate distinct CP types in blends or serially. Other emerging plant virus‐deduced systems include the usually isometric cowpea chlorotic mottle virus (CCMV) with further strikingly altered structures up to "cherrybombs" with protruding nucleic acids. Cartoon strips and pictorial descriptions of major RNA‐based strategies induct the reader into a rare field of nanoconstruction that can give rise to utile soft‐matter architectures for complex tasks. This article is categorized under:Biology‐Inspired Nanomaterials > Protein and Virus‐Based StructuresNanotechnology Approaches to Biology > Nanoscale Systems in BiologyBiology‐Inspired Nanomaterials > Nucleic Acid‐Based Structures [ABSTRACT FROM AUTHOR]

Gold nanoparticles (GNPs), with tunable optical properties, bioinertness, and surface multivalent effect, have been widely explored for biomedical applications. As one classical type of GNPs‐based assemblies, plasmonic gold nanovesicles (GVs), with a hollow cavity, "solid skeleton" composed of GNPs cores and a "soft body" composed of functional polymers, have attracted considerable attention due to their tunable localized surface plasmon resonance, strong surface‐enhanced Raman scattering properties, and high photothermal conversion efficiency. This review summarizes recent advances in biomedical applications for plasmonic GVs. Firstly, the synthesis methods of GVs are mainly including self‐assembly and in situ gold growth methods. Secondly, the classification of GVs is described according to the morphology of GNPs cores. Thirdly, different biomedical applications of GVs are elaborated, including in vitro diagnosis, in vivo imaging, and in vivo therapy. Finally, the challenges and perspectives of GVs are discussed. Gold nanovesicles (GVs), one classical type of gold nanoparticles‐based assemblies, have attracted considerable attention due to their unique plasmonic property in biomedical applications. In this review, synthesis methods, classification, biomedical applications, challenges, and perspectives of GVs are discussed. [ABSTRACT FROM AUTHOR]

C-reactive protein (CRP) is a very important biomarker of infection and inflammation for a number of diseases. Routine CRP measurements with high sensitivity and reliability are highly relevant to the assessment of states of inflammation and the efficacy of treatment intervention, and require the development of very sensitive, selective, fast, robust and reproducible assays. Gold nanoparticles (Au NPs) are distinguished for their unique electrical and optical properties and the ability to conjugate with biomolecules. Au NP-based probes have attracted considerable attention in the last decade in the analysis of biological samples due to their simplicity, high sensitivity and selectivity. Thus, this article aims to be a critical and constructive analysis of the literature of the last three years regarding the advances made in the development of bioanalytical assays based on gold nanoparticles for the in vitro detection and quantification of C-reactive protein from biological samples. Current methods for Au NP synthesis and the strategies for surface modification aiming at selectivity towards CRP are highlighted. [ABSTRACT FROM AUTHOR]