The 50 most-cited articles in The Analyst, 2021

The high-performance liquid chromatography-mass spectrometry (LC-MS) technique is widely applied to routine analysis in many matrices. Despite the enormous application of LC/MS, this technique is subjected to drawbacks called matrix effects (MEs) that could lead to ion suppression or ion enhancement. This phenomenon can exert a deleterious impact on the ionization efficacy of an analyte and subsequently on the important method performance parameters. LC-MS susceptibility to MEs is the main challenge of this technique in the analysis of complex matrices such as biological and food samples. Nowadays, the assessment, estimation, and overcoming of the MEs before developing a method is mandatory in any analysis. Two main approaches including the post-column infusion and post-extraction spike are proposed to determine the degree of MEs. Different strategies can be adopted to reduce or eliminate MEs depending on the complexity of the matrix. This could be done by improving extraction and clean-up methods, changing the type of ionization employed, optimization of liquid chromatography…

A label-free multiplexed electrochemical biosensor based on a gold nanoparticles/graphene quantum dots/graphene oxide (AuNPs/GQDs/GO) modified three-screen-printed carbon electrode (3SPCE) array is successfully constructed to detect miRNA-21, miRNA-155, and miRNA-210 biomarkers for the first time. Redox species (anthraquinone (AQ), methylene blue (MB), and polydopamine (PDA)) are used as redox indicators for anchoring capture miRNA probes, which hybridize with the complementary targets, miRNA-21, miRNA-155, and miRNA-210, respectively. After three target miRNAs are present, the square wave voltammetry (SWV) scan displays three well-separated peaks. Each peak indicates the presence of one miRNA, and its intensity quantitatively correlates with the concentration of the corresponding target analyte. This phenomenon results in the substantial decline of the SWV peak current of the redox probes. The developed AuNPs/GQDs/GO-based biosensor reveals excellent performance for simultaneous miRNA sensing. It offers a wide linear dynamic range from 0.001 to 1000 pM with ultrasensitive low…

, full factorial, central composite, and Box-Behnken are discussed as systematic approaches to optimize electrode fabrication to consider the effects from individual variables and their interactions. Then, the principles of machine learning algorithms, including linear and logistic regressions, neural network, and support vector machine, are introduced. These machine learning models have been implemented to extract complex relationships between chemical structures and their electrochemical properties and to analyze complicated electrochemical data to improve calibration and analyte classification, such as in electronic tongues. Lastly, the future of machine learning and experimental designs in electrochemical sensors is outlined. These chemometric strategies will accelerate the development and enhance the performance of electrochemical devices for point-of-care diagnostics and commercialization.

Forensic investigations are often reliant on physical evidence to reconstruct events surrounding a crime. However, there remains a need for more objective approaches to evidential interpretation, along with rigorously validated procedures for handling, storage and analysis. Chemometrics has been recognised as a powerful tool within forensic science for interpretation and optimisation of analytical procedures. However, careful consideration must be given to factors such as sampling, validation and underpinning study design. This tutorial review aims to provide an accessible overview of chemometric methods within the context of forensic science. The review begins with an overview of selected chemometric techniques, followed by a broad review of studies demonstrating the utility of chemometrics across various forensic disciplines. The tutorial review ends with the discussion of the challenges and emerging trends in this rapidly growing field.

Exosomes are nano-sized extracellular vesicles that serve as a communications system between cells and have shown tremendous promise as liquid biopsy biomarkers in diagnostic, prognostic, and even therapeutic use in different human diseases. Due to the natural heterogeneity of exosomes, there is a need to separate exosomes into distinct biophysical and/or biochemical subpopulations to enable full interrogation of exosome biology and function prior to the possibility of clinical translation. Currently, there exists a multitude of different exosome isolation and characterization approaches which can, in limited capacity, separate exosomes based on biophysical and/or biochemical characteristics. While notable reviews in recent years have reviewed these approaches for bulk exosome sorting, we herein present a comprehensive overview of various conventional technologies and modern microfluidic and nanotechnological advancements towards isolation and characterization of exosome subpopulations. The benefits and limitations of these different technologies to improve their use for distinct…

Diabetes is a group of metabolic conditions resulting in high blood sugar levels over prolonged periods that affects hundreds of millions of patients worldwide. Measuring glucose concentration enables patient-specific insulin therapy, and is essential to reduce the severity of the disease, potential complications, and related mortalities. Recent advances and developments in smartphone-based colorimetric glucose detection systems are discussed in this review. The importance of glucose monitoring, data collection, transfer, and analysis, via non-invasive/invasive methods is highlighted. The review also presents various approaches using 3D-printed materials, screen-printed electrodes, polymer templates, designs allowing multiple glucose analysis, bioanalytes and/or nanostructures for glucose detection. The positive effects of advances in improving the performance of smartphone-based platforms are introduced along with future directions and trends in the application of emerging technologies in smartphone-based diagnostics.

Numerous clinical studies suggest that microRNAs (miRNAs) are indicative biomolecules for the early diagnosis of cancer. This work aims to develop a cost-effective and label-free electrochemical biosensor to detect miRNA-21, a biomarker of breast cancer. An electrochemical sensor is fabricated using a nanocomposite, consisting of graphene (GP), polypyrrole (PPY) and gold nanoparticles (AuNPs), modified onto a screen-printed carbon electrode (SPCE) to improve electron transfer properties and increase the degree of methylene blue (MB) intercalation for signal amplification. The GP/PPY-modified electrode offers good electrochemical reactivity and high dispersibility of AuNPs, resulting in excellent sensor performance. Peak current of the MB redox process, which is proportional to miRNA-21 concentration on the electrode surface, is monitored by differential pulse voltammetry (DPV). Under optimal conditions, this sensor is operated by monitoring the MB signal response due to the amount of hybridization products between miRNA-21 target molecules and DNA-21 probes immobilized on the…

During the past few decades, there has been a growing trend towards the use of smartphone-based analysis systems. This is mainly due to its ubiquity, its increasing computing capacity, its relatively low cost and the ability to acquire and process data at the same time. Furthermore, there are many sensors integrated into a smartphone, for example a complementary metal-oxide semiconductor (CMOS) sensor. A CMOS sensor enables optical analysis for example by using it as a colorimeter, photometer or spectrometer. This review explores the current state-of-the-art smartphone-based optical analysis systems in various areas of application. It is organized into three sections, each of which investigates one class of smartphone-based devices: (i) smartphone-based colorimeters (ii) smartphone-based photo- and spectrometers and (iii) smartphone-based fluorimeters.

Recently, lateral flow assay (LFA) for nucleic acid detection has drawn increasing attention in the point-of-care testing fields. Due to its rapidity, easy implementation, and low equipment requirement, it is well suited for use in rapid diagnosis, food authentication, and environmental monitoring under source-limited conditions. This review will discuss two main research directions of lateral flow nucleic acid tests. The first one is the incorporation of isothermal amplification methods with LFA, which ensures an ultra-high testing sensitivity under non-laboratory conditions. The two most commonly used methodologies will be discussed, namely Loop-mediated Isothermal Amplification (LAMP) and Recombinase Polymerase Amplification (RPA), and some novel methods with special properties will also be introduced. The second research direction is the development of novel labeling materials. It endeavors to increase the sensitivity and quantifiability of LFA testing, where signals can be read and analyzed by portable devices. These methods are compared in terms of limits of detection,…

There has been an explosion of interest in the use of nanomaterials for biosensing applications, and carbonaceous nanomaterials in particular are at the forefront of this explosion. Carbon dots (CDs), a new type of carbon material, have attracted extensive attention due to their fascinating properties, such as small particle size, tunable optical properties, good conductivity, low cytotoxicity, and good biocompatibility. These properties have enabled them to be highly promising candidates for the fabrication of various high-performance biosensors. In this review, we summarize the top-down and bottom-up synthesis routes of CDs, highlight their modification strategies, and discuss their applications in the fields of photoluminescence biosensors, electrochemiluminescence biosensors, chemiluminescence biosensors, electrochemical biosensors and fluorescence biosensors. In addition, the challenges and future prospects of the application of CDs for biosensors are also proposed.

). Selecting which methods to use is not always clear. Herein, we describe the components of RS instruments and how they influence the quality of Raman spectra acquired from bone because signal-to-noise of the acquisition and the accompanying background fluorescence dictate the pre-processing of the Raman spectra. We also describe common methods and challenges in preparing acquired spectra for the determination of matrix properties of bone. This article also serves to provide guidance for the analysis of bone by RS with examples of how methods for pre-processing the Raman signals and for determining properties of bone composition affect RS sensitivity to potential differences between experimental groups. Attention is also given to deconvolution methods that are used to ascertain sub-peak ratios of the amide I band as a way to assess characteristics of collagen type I. We provide suggestions and recommendations on the application of RS to bone with the goal of improving reproducibility across studies and solidify RS as a valuable technique in the field of bone research.

Low glucose levels during exercise may lead to hypoglycemia, which can have grave consequences in diabetic athletes. Mobile colorimetric wearable biosensors that measure glucose levels in sweat are ideal for self-monitoring as they can utilize the camera in smartphones for signal reading. However, colorimetric biosensors proposed thus far have higher limit of detection (LOD) than electrochemical devices, which makes them unsuitable for detecting hypoglycemia. In this manuscript we describe colorimetric wearable biosensors that detect glucose in sweat with an LOD of 0.01 mM and a dynamic range up to 0.15 mM. The devices are made of filter paper and incorporate a sweat volume sensor and a color chart for signal correction. The biosensors do not suffer from interferences originated by delayed sample readings, or differences in bending angle and sample pH. When applied to volunteers performing an exercise routine, sweat glucose levels corrected with sweat volume measurements correlated well with blood glucose measurements performed with a commercial device. The devices are lightweight…

Herein a rapid and sensitive fluorometric bioanalysis platform for mercury(ii) (Hg2+) detection was innovatively developed using ultrathin two-dimensional MXenes (Ti3C2) as fluorescence quencher and Hg2+-induced exonuclease III (Exo III)-assisted target recycling strategy for efficient signal amplification. Initially, fluorophore-labeled single-stranded DNA (FAM-labeled probe) can be easily adsorbed onto the surface of ultrathin Ti3C2 nanosheets by hydrogen bonding and metal chelating interaction, and the fluorescence signal emitted by the FAM-labeled probe is quenched strongly owing to the fluorescence resonance energy transfer between the FAM and ultrathin Ti3C2 nanosheets. Upon sensing the target Hg2+, the protruding DNA fragment at the 3' end of hairpin will hybridize with primer (hairpin-Hg2+-primer), and then further digested by Exo III to produce a probe (nicker). The released target Hg2+ and primer continue to participate in the next recycling, resulting in more hairpin probes becoming nickers. The combination of a large number of nickers and FAM-probe resulted in a…

might be interfered by the nanozyme with photosensibility. (5) Both the quality of DMPO and incubation time are important factors for electron paramagnetic resonance (EPR) measurement. This study will be useful for choosing more suitable in vitro detection methods of SOD-like activity for nanozymes in the future.

Capillary-driven microfluidic devices are of significant interest for on-site analysis because they do not require external pumps and can be made from inexpensive materials. Among capillary-driven devices, those made from paper and polyester film are among the most common and have been used in a wide array of applications. However, since capillary forces are the only driving force, flow is difficult to control, and passive flow control methods such as changing the geometry must be used to accomplish various analytical applications. This study presents several new flow control methods that can be utilized in a laminate capillary-driven microfluidic device to increase available functionality. First, we introduce push and burst valve systems that can stop and start flow. These valves can stop flow for >30 min and be opened by either pressing the channel or inflowing other fluids to the valve region. Next, we propose flow control methods for Y-shaped channels that enable more functions. In one example, we demonstrate the ability to accurately control concentration to create laminar,…

(Mn(ii)). The lowest detectable concentration and linearity for the five metal ions allow the application of this device for the determination of heavy metal ions in various water samples. The sensor showed high selectivity and efficiency for simultaneous determination of Cu(ii), Co(ii), Ni(ii), Hg(ii), and Mn(ii) in drinking, tap, and pond water samples on a single device and detection with the naked eye. The results illustrated that the proposed sensor showed good accuracy and precision agreement with the standard ICP-OES method.

Acute myocardial infarction (AMI) is the main cause of death from cardiovascular diseases. Thus, early diagnosis of AMI is essential for the treatment of irreversible damage from myocardial infarction. Traditional electrocardiograms (ECG) cannot meet the specific detection of AMI. Cardiac troponin I (cTnI) is the main biomarker for the diagnosis of myocardial infarction, and the detection of cTnI content has become particularly important. In this review, we introduced and compared the advantages and disadvantages of various cTnI detection methods. We focused on the analysis and comparison of the main indicators and limitations of various cTnI biosensors, including the detection range, detection limit, specificity, repeatability, and stability. In particular, we pay more attention to the application and development of electrochemical biosensors in the diagnosis of cardiovascular diseases based on different biological components. The application of electrochemical microfluidic chips for cTnI was also briefly introduced in this review. Finally, this review also briefly discusses the…

are labeled and traced using specific fluorescent probes, therefore helping us to explore their physiological function and pathological change. Among them, intracellular pH value is an important factor that governs biological processes, generally ∼7.2. Furthermore, specific organelles within cells possess unique acid-base homeostasis, involving the acidic lysosomes, alkalescent mitochondria, and neutral endoplasmic reticulum and Golgi apparatus, which undergo various physiological processes such as intracellular digestion, ATP production, and protein folding and processing. In this review, recently reported fluorescent probes targeted toward the lysosomes, mitochondria, endoplasmic reticulum, Golgi apparatus, and cytoplasm for sensing pH change are discussed, which involves molecular structures, fluorescence behavior, and biological applications.

Cell separation has consistently been a pivotal technology of sample preparation in biomedical research. Compared with conventional bulky cell separation technologies applied in the clinic, cell separation based on microfluidics can accurately manipulate the displacement of liquid or cells at the microscale, which has great potential in point-of-care testing (POCT) applications due to small device size, low cost, low sample consumption, and high operating accuracy. Among various microfluidic cell separation technologies, inertial microfluidics has attracted great attention due to its simple structure and high throughput. In recent years, many researchers have explored the principles and applications of inertial microfluidics and developed different channel structures, including straight channels, curved channels, and multistage channels. However, the recently developed multistage channels have not been discussed and classified in detail compared with more widely discussed straight and curved channels. Therefore, in this review, a comprehensive and detailed review of recent progress…

Biogenic amines are involved in physiological roles in living organisms, but their excessive production or intake can induce undesired toxicological effects. As biogenic amines can be found in the process of food spoilage, they are considered an indicator of food quality and freshness, and their detection is of crucial importance in food safety. In this contribution, we report the fast and direct colorimetric and fluorometric sensing of biogenic amines by means of a dinuclear Zn(ii) Schiff-base complex. The selective and sensitive detection involves the formation of stable adducts between the dinuclear complex, acting as the Lewis acidic molecular tweezer, and biogenic di- or polyamines. The selectivity towards biogenic amines, even in the presence of common aliphatic, primary, secondary, or tertiary monoamines, heterocyclic amines, and amino acids, is demonstrated by competitive experiments. The quantitation of histamine in a fish matrix is easily achieved using a standard extraction procedure followed by simple colorimetric or fluorometric measurements.

The use of infrared spectroscopy to augment decision-making in histopathology is a promising direction for the diagnosis of many disease types. Hyperspectral images of healthy and diseased tissue, generated by infrared spectroscopy, are used to build chemometric models that can provide objective metrics of disease state. It is important to build robust and stable models to provide confidence to the end user. The data used to develop such models can have a variety of characteristics which can pose problems to many model-building approaches. Here we have compared the performance of two machine learning algorithms - AdaBoost and Random Forests - on a variety of non-uniform data sets. Using samples of breast cancer tissue, we devised a range of training data capable of describing the problem space. Models were constructed from these training sets and their characteristics compared. In terms of separating infrared spectra of cancerous epithelium tissue from normal-associated tissue on the tissue microarray, both AdaBoost and Random Forests algorithms were shown to give excellent…

exosomes per mL with a detection limit of 193 exosomes per mL was obtained. Through monitoring five proteins (CD81, CEA, EpCAM, CD147, and AFP) of exosomes from three different cell lines (A549, HEK293, and HepG2), a significant difference in marker expression levels was observed in different cell lines. Therefore, this method has good prospects in exosome-based tumor marker detection and cancer diagnosis.

Molecularly imprinted polymers (MIPs) have numerous applications in the sensing field, the detection/recognition of virus, the structure determination of proteins, drug delivery, artificial/biomimetic antibodies, drug discovery, and cell culturing. There are lots of conventional methods routinely deployed for the analysis/detection of viral infections and pathogenic viruses, namely enzyme immunoassays, immunofluorescence microscopy, polymerase chain reaction (PCR) and virus isolation. However, they typically suffer from higher costs, low selectivity/specificity, false negative/positive results, time consuming procedures, and inherent labor intensiveness. MIPs offer promising potential for viral recognition/detection with high target selectivity, sensitivity, robustness, reusability, and reproducible fabrication. In terms of virus detection, selectivity and sensitivity are critical parameters determined by the template; additionally, the analytical detection and evaluation of viruses must have considerably low detection limits. The virus-imprinted polymer-based innovative strategies…

was presented. To sum up, our study not only demonstrates the differentiation ability of this nanozyme sensor array but also gives hints for using nanozyme sensor arrays for further applications.

Surface-enhanced Raman spectroscopy (SERS) is an emerging analytical technique for chemical analysis, which is favourable due to its combination of short measurement time, high sensitivity and molecular specificity. However, the application of SERS is still limited, largely because in real samples the analyte is often present in a complex matrix that contains micro/macro particles that block the probe laser, as well as molecular contaminants that compete for the enhancing surface. Here, we show a simple and scalable spray-deposition technique to fabricate SERS-active paper substrates which combine sample filtration and enhancement in a single material. Unlike previous spray-deposition methods, in which simple colloidal nanoparticles were sprayed onto solid surfaces, here the colloidal nanoparticles are mixed with hydroxyethyl cellulose (HEC) polymer before application. This leads to significantly improved uniformity in the distribution of enhancing particles as the film dries on the substrate surface. Importantly, the polymer matrix also protects the enhancing particles from…

copies of SARS-CoV-2 RNA. Clinical samples were tested using our RT-LAMP protocol as well as by conventional RT-qPCR, demonstrating comparable performance to the CDC SARS-CoV-2 RT-qPCR assay. The methodology described in this study represents a simple, rapid, and accurate method for rapid molecular diagnostics of COVID-19 in a disposable microdevice, ideal for point-of-care testing (POCT) systems.

Innovative methodological approaches are needed to conduct human health and environmental risk assessments on a growing number of marketed chemicals. Metabolomics is progressively proving its value as an efficient strategy to perform toxicological evaluations of new and existing substances, and it will likely become a key tool to accelerate chemical risk assessments. However, additional guidance with widely accepted and harmonized procedures is needed before metabolomics can be routinely incorporated in decision-making for regulatory purposes. The aim of this review is to provide an overview of metabolomic strategies that have been successfully employed in toxicity assessment as well as the most promising workflows in a regulatory context. First, we provide a general view of the different steps of regulatory toxicology-oriented metabolomics. Emphasis is put on three key elements: robustness of experimental design, choice of analytical platform, and use of adapted data treatment tools. Then, examples in which metabolomics supported regulatory toxicology outputs in different scenarios…

Progress in real-time, simultaneous in vivo detection of multiple neurotransmitters will help accelerate advances in neuroscience research. The need for development of probes capable of stable electrochemical detection of rapid neurotransmitter fluctuations with high sensitivity and selectivity and sub-second temporal resolution has, therefore, become compelling. Additionally, a higher spatial resolution multi-channel capability is required to capture the complex neurotransmission dynamics across different brain regions. These research needs have inspired the introduction of glassy carbon (GC) microelectrode arrays on flexible polymer substrates through carbon MEMS (C-MEMS) microfabrication process followed by a novel pattern transfer technique. These implantable GC microelectrodes provide unique advantages in electrochemical detection of electroactive neurotransmitters through the presence of active carboxyl, carbonyl, and hydroxyl functional groups. In addition, they offer fast electron transfer kinetics, capacitive electrochemical behavior, and wide electrochemical window. Here,…

Matrix metalloproteinase-1 (MMP-1) is associated with many types of cancers, including oral, colorectal, and brain cancers. This paper describes the fabrication of an MMP-1 immunosensor based on a gold nanoparticle/polyethyleneimine/reduced graphene oxide (AuNP/PEI/rGO)-modified disposable screen-printed electrode (SPE). A microwave-assisted single-step method was employed for the simultaneous reduction of gold and graphene oxide in a PEI environment to avoid AuNP agglomeration. The crystal structure, chemical composition, optical properties, and interior morphology of the materials were probed by X-ray diffraction, Raman spectroscopy, UV-visible spectrometry, and transmission electron microscopy techniques. To assemble a label-free MMP-1 immunosensor layer-by-layer, 3-mercaptopropionic acid was utilized due to its strong sulfur-gold bonding ability, and its tail end was attached to a carboxyl group, allowing the MMP-1 antibody (anti-MMP-1) to be subsequently cross-linked using the traditional N-(3-dimethylaminopropyl) and N' ethylcarbodiimide hydrochloride method. Differential…

can be detected with low limit of detection (0.038 μM) in a linear range of 0 to 3 mM. The probe is expected to be able to detect ammonia in the environment.

Cell viability is a physiological status connected to cell membrane integrity and cytoplasmic topography, which is profoundly important for fundamental biological research and practical biomedical applications. A conventional method for assessing cell viability is through cell staining analysis. However, cell staining involves laborious and complicated processing procedures and is normally cytotoxic. Intrinsic cellular phenotypes thus provide new avenues for measuring cell viability in a stain-free and non-toxic manner. In this work, we present a label-free non-destructive impedance-based approach for cell viability assessment by simultaneously characterizing multiple electrical cellular phenotypes in a high-throughput manner (>1000 cells per min). A novel concept called the complex opacity spectrum is introduced for improving the discrimination of live and dead cells. The analysis of the complex opacity spectrum leads to the discovery of two frequency ranges that are optimized for characterizing membranous and cytoplasmic electrical phenotypes. The present impedance-based approach…

Chiral drugs are drugs with chiral or asymmetric centres in their molecular structure. Different enantiomers of the same chiral drug have noticeably different pharmacological activities and pharmacokinetic properties. However, its distinction has been perplexing scholars for many years in the qualitative and quantitative detection of antagonistic drugs. Conventional detection methods, such as polarimetry, circular dichroism, and high-performance liquid chromatography, are time consuming, cause sample loss and have cumbersome operations, and they can be applied only to the sampling method. In this paper, we propose a fast, accurate, qualitative and quantitative method for the study of chiral drugs based on linearly polarized terahertz (THz) spectroscopy and imaging technology. Taking ibuprofen as an example, based on the THz absorption spectra of the enantiomers RS-ibuprofen, (R)-(-)-ibuprofen, and (S)-(+)-ibuprofen, their characteristic peak frequencies, peak amplitude differences and peak area differences were extracted to qualitatively and quantitatively distinguish and identify…

The human body comprises rich populations of cells, which are arranged into tissues and organs with diverse functionalities. These cells exhibit a broad spectrum of phenotypes and are often organized as a heterogeneous but sophisticatedly regulated ecosystem - tissue microenvironment, inside which every cell interacts with and is reciprocally influenced by its surroundings through its life span. Therefore, it is critical to comprehensively explore the cellular machinery and biological processes in the tissue microenvironment, which is best exemplified by the tumor microenvironment (TME). The past decade has seen increasing advances in the field of spatial proteomics, the main purpose of which is to characterize the abundance and spatial distribution of proteins and their post-translational modifications in the microenvironment of diseased tissues. Herein, we outline the achievements and remaining challenges of mass spectrometry-based tissue spatial proteomics. Exciting technology developments along with important biomedical applications of spatial proteomics are highlighted. In…

A new fluorescent probe, 3-(benzo[ d ]thiazol-2-yl)-5-bromosalicylaldehyde- 4 N -phenyl thiosemicarbazone (BTT), for ratiometric sensing of Zn 2+ ions in methanol/HEPES buffer solution (3 : 2, pH = 7.4) is reported in this paper.

. Finally, we discuss the limitations associated with the existing applications and potential breakthroughs that can be achieved in this field in the future.

M, respectively. The Cu FMA colorimetric sensor can be applied for detecting and measuring glucose and epinephrine in human serum samples. This work paves the way for Cu MOFs to be used as the basis for rational regulation of the activity of dual nanozymes and for multifunctional applications under completely independent conditions.

Since the discovery of the enzyme-like activities of nanomaterials, the study of nanozymes has become one of the most popular research frontiers of diverse areas including biosensors. DNA also plays a very important role in the construction of biosensors. Thus, the idea of combined applications of nanozymes with DNA (DNA-nanozyme) is very attractive for the development of nanozyme-based biosensors, which has attracted considerable interest of researchers. To date, many sensors based on DNA-functionalized or templated nanozymes have been reported for the detection of various targets and highly accelerated the development of nanozyme-based sensors. In this review, we summarize the main applications and advances of DNA-nanozyme-based sensors. Additionally, perspectives and challenges are also discussed at the end of the review.

The pandemic outbreak of the 2019 coronavirus disease (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still spreading rapidly and poses a great threat to human health. As such, developing rapid and accurate immunodiagnostic methods for the identification of infected persons is needed. Here, we proposed a simple but sensitive on-site testing method based on spike protein-conjugated quantum dot (QD) nanotag-integrated lateral flow immunoassay (LFA) to simultaneously detect SARS-CoV-2-specific IgM and IgG in human serum. Advanced silica-core@dual QD-shell nanocomposites (SiO2@DQD) with superior luminescence and stability were prepared to serve as fluorescent nanotags in the LFA strip and guarantee high sensitivity and reliability of the assay. The performance of the SiO2@DQD-strip was fully optimized and confirmed by using 10 positive serum samples from COVID-19 patients and 10 negative samples from patients with other respiratory diseases. The practical clinical value of the assay was further evaluated by testing 316 serum samples…

A phenothiazine-based fluorescent probe features high selectivity and sensitivity, low cytotoxicity and reliability at physiological pH, enabling the detection of H 2 S in biosystems and monitoring H 2 S produced in the foods spoilage process.

The discovery of electroactive nano-biomaterials and the development of flexible electrodes have increased the interest in applications of integrated electrochemical lateral flow immunoassays (eLFIAs), which integrate electrochemical nanotags and flexible electrodes on test strips that can easily detect small biomolecules. Compared with colorimetric, optical, magnetic and other highly sensitive detection methods, the electrochemical detection technique is well developed with high sensitivity, selectivity and repeatability. Moreover, the increasing compatibility of interfaces with miniature potentiometers has allowed electrochemical sensors to become more integrated, automated and intelligent, highlighting their huge potential in future developments. This review discusses the relevant eLFIA research over the past 20 years. The basic principles, electrode assemblies, electrochemical labeling strategies and electrical signal detection methods are summarized and analyzed in detail. Finally, perspectives on the current challenges facing eLFIA and its future outlook are presented.

Currently, the 2019 novel coronavirus (2019-nCoV) is drastically affecting 214 countries, causing severe pneumonia in patients, which has resulted in lockdown being implemented in several countries to stop its local transmission. Considering this, the rapid screening and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; 2019-nCoV) play an essential role in the diagnosis of COVID-19, which can minimize local transmission and prevent an epidemic. Due to this public health emergency, the development of ultra-fast reliable diagnostic kits is essential for the diagnosis of COVID-19. Recently, molecular biology and nanotechnology based analytical methods have proven to be promising diagnostic tools for the rapid screening of 2019-nCoV with high accuracy and precision. The main aim of this review is to provide a retrospective overview on the molecular biology tools (reverse transcription polymerase chain reaction (RT-PCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP)) and nanotechnology based analytical tools (enzyme-linked…

Heavy metals are the main pollutants present in aquatic environments and their presence in human organisms can lead to many different diseases. While many methods exist for analysis, colorimetric and electrochemistry are particularly attractive for on-site analysis and their integration on a single platform can improve multiplexed metals analysis. This report describes for the first time a "plug-and-play" (PnP) assembly for coupling a microfluidic paper-based device (μPAD) and a screen-printed electrochemical paper-based device (ePAD) using a vertical and reversible foldable mechanism for multiplexed detection of Fe, Ni, Cu, Zn, Cd and Pb in river water samples. The integration strategy was based on a reversible assembly, allowing the insertion of a pretreatment zone to minimize potential chemical interfering agents and providing a better control of the aspirated sample volume as well as to a lower sample evaporation rate. In comparison with lateral flow and electrochemical assays performed using independent devices, the integrated prototype proved that the reversible coupling…

Sensitive, reliable and cost-effective detection of pathogens has wide ranging applications in clinical diagnostics and therapeutics, water and food safety, environmental monitoring, biosafety and epidemiology. Nucleic acid amplification tests (NAATs) such as PCR and isothermal amplification methods provide excellent analytical performance and significantly faster turnaround times than conventional culture-based methods. However, the inherent cost and complexity of NAATs limit their application in resource-limited settings and the developing world. To help address this urgent need, we have developed a sensitive method for nucleic acid analysis based on padlock probe rolling circle amplification (PLRCA), nuclease protection (NP) and lateral flow detection (LFA), referred to as PLAN-LFA, that can be used in resource-limited settings. The assay involves solution-phase hybridization of a padlock probe to target, sequence-specific ligation of the probe to form a circular template that undergoes isothermal rolling circle amplification in the presence of a polymerase and a labeled probe…

The ability to visually detect low numbers of Salmonella in food samples is highly valuable but remains a challenge. Here we present a novel platform for ultrasensitive and visual detection of Salmonella Typhimurium by integrating the bio-barcode immunoassay (BCA), recombinase polymerase amplification (RPA), and CRISPR-Cas12a cleavage in a single reaction system (termed as BCA-RPA-Cas12a). In the system, the target bacteria were separated by immunomagnetic nanoparticles and labeled with numerous barcode AuNPs, which carry abundant bio-barcode DNA molecules to amplify the signal. Afterwards, the bio-barcode DNA molecules were amplified by RPA and subsequently triggered the cleavage activity of Cas12a to generate the fluorescence signal. Due to this triplex signal amplification, the BCA-RPA-Cas12a system can selectively detect Salmonella Typhimurium at the single-digit level with the naked eye under blue light within 60 min. Meanwhile, this novel platform was successfully applied to detect Salmonella Typhimurium in spiked milk samples with a similar sensitivity and satisfactory…

Surface enhanced Raman scattering (SERS) is a technique that demonstrates a number of advantages for the rapid, specific and sensitive detection of pathogenic microorganisms. In this review, an overview of label-free and label-based SERS approaches, including microfluidics, nucleic acid detection and immunoassays, for the multiplexed detection of pathogenic bacteria and viruses from the last decade will be discussed, as well as their transition into promising point-of-use detection technologies in industrial and medical settings.

Osteosarcoma is the most frequent primary bone cancer, particularly among children and adolescents. The early diagnosis of osteosarcoma is significant for timely clinical treatment to reduce the mortality of patients. Exosomes play a significant role in intercellular communication and serve as promising biomarkers in liquid biopsy for the diagnosis and monitoring of tumors. Herein, we report the utility of surface-enhanced Raman scattering (SERS) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for rapid identification of osteosarcoma. We firstly profiled the intrinsic SERS signals and MALDI-TOF mass fingerprints of different subgroups of extracellular vesicles (EVs) and the corresponding cells, demonstrating that the SERS signals and MALDI-TOF mass spectra of exosomes from different types of cells were more discriminative compared to those of large and medium EVs and the cells themselves. Then, we characterized plasma-derived exosomes of 15 osteosarcoma patients and 15 healthy volunteers using SERS and MALDI-TOF MS, revealing…

Aptamers, which are called chemical antibodies for their high affinity and specificity to targets, have great potential as analytical tools to detect pesticides. In this work, a DNA aptamer for thiamethoxam was isolated by an improved SELEX (systematic evolution of ligands by exponential enrichment) strategy, in which the ssDNA library was fixed on streptavidin-agarose beads through a short biotin labeled complementary strand. After 13 rounds of selection, the random ssDNA pool was successfully enriched. Three sequences were chosen as aptamer candidates through sequencing and analysis and were transformed into fluorescent probes to evaluate their interactions with thiamethoxam. A fluorescent turn-on aptasensor for thiamethoxam based on the best aptamer (FAM-Thi13) and a short quenching strand were further designed and showed a quantitative linear range from 10 to 1000 nM with a detection limit of 1.23 nM for thiamethoxam. Molecular docking and molecular dynamics were used to investigate the binding site of the main probe of the aptasensor (FAM-Thi13) and thiamethoxam. Satisfactory…

O and is more easily detached from water and compatible with the long-chain DES phase. The motion of EV and MV on the phase interface of DES and water was calculated to further analyze from the molecular level. At the same time, EV tends to move into the DES phase. In summary, the excellent extraction ability of DES for TPM is verified through experiments and simulations, providing solid theoretical support in terms of separation in other fields.

We report on the development of surface plasmon resonance (SPR) sensors and matching ELISAs for the detection of nucleocapsid and spike antibodies specific against the novel coronavirus 2019 (SARS-CoV-2) in human serum, plasma and dried blood spots (DBS). When exposed to SARS-CoV-2 or a vaccine against SARS-CoV-2, the immune system responds by expressing antibodies at levels that can be detected and monitored to identify the fraction of the population potentially immunized against SARS-CoV-2 and support efforts to deploy a vaccine strategically. A SPR sensor coated with a peptide monolayer and functionalized with various sources of SARS-CoV-2 recombinant proteins expressed in different cell lines detected human anti-SARS-CoV-2 IgG antibodies in clinical samples. Nucleocapsid expressed in different cell lines did not significantly change the sensitivity of the assays, whereas the use of a CHO cell line to express spike ectodomain led to excellent performance. This bioassay was performed on a portable SPR instrument capable of measuring 4 biological samples within 30 minutes of…

Clostridium difficile (C. diff) infection is one of the most contagious diseases associated with high morbidity and mortality rates in hospitalised patients. Accurate diagnosis can slow its spread by determining the most effective treatment. Herein, we report a novel testing platform as a proof-of-concept for the selective, sensitive, rapid and cost-effective diagnosis of C. diff infection (CDI) based on a duplex measurement. This was achieved by detecting two specific biomarkers, surface layer protein A (SlpA) and toxin B (ToxB), using a surface enhanced Raman scattering-based lateral flow assay (SERS-based LFA). The simultaneous duplex detection of SlpA with ToxB has not been described for the clinical diagnosis of CDI previously. The SlpA biomarker "AKDGSTKEDQLVDALA" was first reported by our group in 2018 as a species-specific identification tool. The second biomarker, ToxB, is the essential virulence biomarker of C. diff pathogenic strains and is required to confirm true infection pathogenicity. The proposed SERS-based LFA platform enabled rapid duplex detection of SlpA and…