We present a novel "on-off", cost-effective, rapid electrochemical aptasensor combined with a microfluidics cartridge system for the detection of Δ9-THC (Δ9-tetrahydrocannabinol) in human saliva via differential pulse voltammetry. The assay relied on the competitive binding between the Δ9-THC and a soluble redox indicator methylene blue, using an aptamer selected via FRELEX. We found that the aptasensor can detected 1 nM of Δ9-THC in PBS in a three-electrode cell system, while the sensitivity and both the dissociation constant (Kd) and association constant (Kb) were dependent on the aptamer density. The aptamer also showed great affinity towards Δ9-THC when tested against cannabinol and cannabidiol. The same limit of detection of 1 nM in PBS was achieved in small volume samples (∼60 μL) using the aptamer-modified gold screen-printed electrodes combined with the microfluidic cartridge setup, however, the presence of 10% raw human saliva had a negative effect which manifested in a 10-fold increase in the LOD due to interfering elements. Filtering the saliva, improved the tested volume to 50% and the LOD to 5 nM of Δ9-THC which is lower than the concentrations associated with impairment (6.5-32 nM). The aptasensor showed a good storage capability up to 3 days, however, the reusability significantly dropped from 10 cycles (freshly prepared) to 5 cycles. The results clearly demonstrate the feasibility of the aptasensor platform with the microfluidics chamber towards a point-of-care testing application for the detection of Δ9-THC in saliva.

For point-of-care testing (POCT), coupling isothermal nucleic acid amplification schemes (e.g., recombinase polymerase amplification, RPA) with lateral flow assay (LFA) readout is an ideal platform, since such integration offers both high sensitivity and deployability. However, isothermal schemes typically suffers from non-specific amplification, which is difficult to be differentiated by LFA and thus results in false-positives. Here, we proposed an accurate POCT platform by specific recognition of target amplicons with peptide nucleic acid (PNA, assisted by T7 Exonuclease), which could be directly plugged into the existing RPA kits and commercial LFA test strips. With SARS-CoV-2 as the model, the proposed method (RPA-TeaPNA-LFA) efficiently eliminated the false-positives, exhibiting a lowest detection concentration of 6.7 copies/μL of RNA and 90 copies/μL of virus. Using dual-gene (orf1ab and N genes of SARS-CoV-2) as the targets, RPA-TeaPNA-LFA offered a high specificity (100%) and sensitivity (RT-PCR Ct < 31, 100%; Ct < 40, 71.4%), and is valuable for on-site screening or self-testing during isolation. In addition, the dual test lines in the test strips were successfully explored for simultaneous detection of SARS-CoV-2 and H1N1, showing great potential in response to future pathogen-based pandemics.

Delayed diagnosis of cancer-causing death is a worldwide concern. General diagnosis methods are invasive, time-consuming, and operation complicated, which are not suitable for preliminary screening. To address these challenges, the sensing platform based on immune scaffold and fully automated saliva analyzer (FASA) was proposed for oral cancer screening for the first time by non-invasive detection of Cyfra21-1 in saliva. Through one-step synthesis method with unique covalent and electrostatic adsorption strategy, AuNPs@HRP@FeMOF immune scaffold features multiple functions including antibody carrier, catalytic activity, and signal amplification. Highly integrated FASA with the immune scaffold provides automatic testing to avoid false-positive results and reduce pretreatment time without any user intervention. Compared with the commercial analyzer, FASA has comparable performance for Cyfra21-1 detection with a detection range of 3.1-50.0 ng/mL and R2 of 0.971, and superior features in full automation, high integration, time saving and low cost. Oral cancer patients could be distinguished accurately by the platform with an excellent correlation (R2 of 0.904) and average RSD (5.578%) without sample dilution. The proposed platform provides an effective and promising tool for cancer screening in point-of-care applications, which can be further extended for biomarker detection in universal body fluids, disease screening, prognosis review and homecare monitoring.

Biosensors based on graphene field-effect transistors have become a promising tool for detecting a broad range of analytes. However, their performance is substantially affected by the functionalization protocol. In this work, we use a controlled in-vacuum physical method for the covalent functionalization of graphene to construct ultrasensitive aptamer-based biosensors (aptasensors) able to detect hepatitis C virus core protein. These devices are highly specific and robust, achieving attomolar detection of the viral protein in human blood plasma. Such an improved sensitivity is rationalized by theoretical calculations showing that induced polarization at the graphene interface, caused by the proximity of covalently bound molecular probe, modulates the charge balance at the graphene/aptamer interface. This charge balance causes a net shift of the Dirac cone providing enhanced sensitivity for the attomolar detection of the target proteins. Such an unexpected effect paves the way for using this kind of graphene-based functionalized platforms for ultrasensitive and real-time diagnostics of different diseases.

Aβ oligomers have been widely accepted as significant biomarkers for Alzheimer's disease (AD) detection, monitoring, and therapy since they are highly correlated with AD development. In this work, an electrochemical array-based sensing platform was successfully built using a group of functionalized graphene with different physicochemical features. Since the electro-insulated Aβ peptide species severely interfered with the electron transport on the electrode surface, the presence of Aβ led to a significant change in the electrochemical impedance signal. The resulting variety of the impedance was then classified and processed by linear discriminant analysis. The constructed sensing platform can discriminate different Aβ forms, the mixture of various Aβ forms, and different ratios of Aβ42 to Aβ40 with 100% accuracy by only the combination of dual probes. Furthermore, it also exhibited excellent performance for screening Aβ inhibitors and metal chelators. The strategy utilizes the infinitesimal general discrepancy instead of specific biomarker recognition, exhibiting the advantage of no requirement to know the exact information about the specific ligand and receptor in advance, which is promising to be widened for the other biosensing detection fields.

During industrial fermentation, microbial cell factories are usually confronted with environmental or metabolic stresses, leading to the imbalance of intracellular redox and the reduction of cell metabolic capacity. Here, we constructed the genetically encoded redox biosensor system (RBS) based on redox-sensitive fluorescent proteins to detect redox metabolites, including reactive oxygen species (ROS), oxidized glutathione, NADH, and NADPH in Saccharomyces cerevisiae. The functional biosensors were quantitatively characterized and the orthogonal redox biosensor system (oRBS) was designed for detecting multiple redox metabolites. Furthermore, the compartment targeted redox biosensor system (ctRBS) was constructed to detect ROS and NADPH, revealing the distribution and spatiotemporal dynamics of ROS in yeast under various stress conditions. As a proof-of-concept, RBS was applied to evaluate the redox states of engineered yeast with stress resistance and heterogenous triterpene synthesis in vivo, elucidating the redox balance significantly affecting the growth and production phenotypes. The RBS in this study allowed the exploration of the diversity of compartmental redox state and real-time monitoring of the production process of yeast, providing a reliable and effective approach for accurate and in-depth profiling of bottlenecks of yeast cell factories.

Monitoring Interleukin 10 (IL-10) is essential for understanding the vast responses of T-cells in cancer, autoimmunity, and internal homeostasis after physical stress. However, current diagnostic methods are complex and more focused on medical screening rather than point-of-care monitoring. Biosensors based on graphene's conductivity and flexibility are attractive to offer simple single-use and reduced handling. However, oxidation of its carbon lattice to develop functional moieties for biomolecule immobilization cuts down its electronic conductivity potential. In this work, the authors present a microfluidic lab-on-chip device for simple impedimetric monitoring of IL-10 based on graphene foam (GF) flexible electrodes. Graphene's structure was maintained by employing π-π non-covalent functionalization with pyrene carboxylic acid (PCA). Impedimetric measurements could be performed in low ionic strength phosphate-buffered saline (LI-PBS). The PCA-antibody modification showed to endure the incubation, measurement, and washing processes performed in the microfluidic device. Electrode modification and measurements were characterized by, electrochemical impedance spectroscopy (EIS), contact angle, and scanning electron microscopy. From the contact angle results, we found that the wettability of the graphene surface increased gradually after each modification step. Detection measurements performed in the 3D-printed microfluidic device showed a linear response between 10 fg/mL to 100 fg/mL with a limit of detection (LOD) of 7.89 fg/mL in artificial saliva. With these features, the device was used to quantify IL-10 samples by the standard addition method for 10 fg and 50 fg with recoveries between 82% and 99%. Specificity was evaluated towards interleukin 6, TNF-⍺ and bovine serum albumin.

Vancomycin (VCM) causes poisoning symptoms at high concentrations; thus, therapeutic drug monitoring is recommended to measure and control blood levels regularly. However, blood analysis at regular intervals does not allow knowing the detailed temporal change in concentration. To address this challenge, we developed a long-range surface plasmon (LRSP) aptasensor for measuring VCM label-free and real-time by combining a sensitive LRSP sensor and a peptide aptamer with a VCM recognition site. First, three different biosensors for VCM were compared. One was prepared by immobilizing the peptide aptamer directly on (Direct-Apt) or via a self-assembled monolayer (SAM) on a gold surface (SAM-Apt). The other used anti-VCM antibodies immobilized on a gold surface via the SAM (SAM-Ab). The Direct-Apt showed larger sensor output to VCM than the other biosensors. The dynamic range for VCM was 0.78-100 μM, including the therapeutic range (6.9-13.8 μM). The Direct-Apt also showed the sensor output only from VCM among four different antibiotics, demonstrating the high selectivity for VCM. The VCM captured by the aptamer could be removed by rinsing with phosphate-buffered saline. The measurement was rapid, with 72- and 77-sec response and recovery times, allowing not only repeated but also real-time measurements. Finally, the Direct-Apt in 20% serum solutions showed comparable sensitivity to VCM in the buffer solution, indicating high capability for real-sample.

Accurate COVID-19 screening via molecular technologies is still hampered by bulky instrumentation, complicated procedure, high cost, lengthy testing time, and the need for specialized personnel. Herein, we develop point-of-care upconversion luminescence diagnostics (PULD), and a streamlined smartphone-based portable platform facilitated by a ready-to-use assay for rapid SARS-CoV-2 nucleocapsid (N) gene testing. With the complementary oligo-modified upconversion nanoprobes and gold nanoprobes specifically hybridized with the target N gene, the luminescence resonance energy transfer effect leads to a quenching of fluorescence intensity that can be detected by the easy-to-use diagnostic system. A remarkable detection limit of 11.46 fM is achieved in this diagnostic platform without the need of target amplification, demonstrating high sensitivity and signal-to-noise ratio of the assay. The capability of the developed PULD is further assessed by probing 9 RT-qPCR-validated SARS-CoV-2 variant clinical samples (B.1.1.529/Omicron) within 20 min, producing reliable diagnostic results consistent with those obtained from a standard fluorescence spectrometer. Importantly, PULD is capable of identifying the positive COVID-19 samples with superior sensitivity and specificity, making it a promising front-line tool for rapid, high-throughput screening and infection control of COVID-19 or other infectious diseases.

Methylene blue (MB) is a common multifunctional indicator, which can be applied as a quencher for electrochemiluminescence (ECL) analysis as well as a classical redox probe. Although it is relatively prevalent for MB to study the mechanism with Ru-based luminophores in ECL systems, there are few studies on the effects between MB and co-reactants. In this work, we proposed the first investigation of MB on the luminophore and co-reactant of the self-enhanced ECL composites (nitrogen-doped graphene quantum dots on Ru(bpy)32+-doped silica nanoparticles, NGQDs-Ru@SiO2), respectively. The relatively narrow ECL spectrum of luminophore (Ru@SiO2) and the suitable ultraviolet-visible absorption spectrum of MB led to the ECL resonance energy transfer between them, meanwhile the appropriate energy levels among them facilitated the electron transfer, resulting in a decreased ECL signal (quench mode I). Additionally, the co-reactant (NGQDs) was prone to π-π conjugation with MB due to its abundant π-electrons, which reduced the concentration of NGQDs' intermediates and triggered a weakened ECL signal (quench mode II). Therefore, the dual-quenching effects are ingeniously integrated and designed in one ECL-electrochemical (ECL-EC) ratiometric aptasensor for zearalenone detection, for demonstrating its efficacy in enhancing the sensitivity, which is 4.8-fold higher than Ru@SiO2 alone. This innovative ratiometric aptasensor achieved a relatively wide linear range from 1.0 × 10-15 to 5.0 × 10-8 g mL-1, and obtained a low detection limit of 8.5 × 10-16 g mL-1. Our proposed dual-quenching interactions between MB and NGQDs-Ru@SiO2 will open a new prospective for ECL-EC ratiometric aptasensor, which further broaden the application in sensitive and precise analysis of mycotoxins.

As a diagnostic biomarker, the detection of circular RNA (circRNA) is vital for the early screening of bladder cancer. Usually, the low abundance of circRNA in clinic samples results in the necessarily complicated extraction before detection. In this work, a tetrahedron supported CRISPR/Cas13a cleavage has been explored for direct electrochemical detection of circRNA in urine from bladder cancer. CRISPR/Cas13a system has been reasonably designed to recognize the characteristic back-splice junction site of circRNA. The activated CRISPR/Cas13a by circRNA can cleave uracil bases composed of DNA tetrahedron immobilized on the surface of gold electrode, resulting in the breakage of DNA tetrahedron and the release of electrochemical active molecule methylene blue. By virtue of highly catalytic efficiency of CRISPR/Cas13a and rigid structure of tetrahedron, the developed electrochemical biosensor can directly detect circRNA in 25 μL urine sample with the lowest detection limit of 0.089 fM and linear detection range from 10 fM to 50 nM in less than 10 min, so as to avoid complicated extraction process and benefit for on-site detection.

Quantitative immunodiagnosis is one of the most commonly used methods for in vitro diagnostics. Various bioanalytical methods have been developed to quantitatively diagnose immune analytes; however, they require blood dilution pretreatment, reaction mixing, complicated experimental steps, and can cause diagnostic errors due to the hook effect. To address this issue, we introduced a simple immunoassay based on carbon nanoparticles (CNPs). The assay was designed to have high flexibility for use in various in vitro diagnostic devices by constructing a soluble solid-phase immune sensor with high solubility using antibody-conjugated CNPs and polymer materials. Excellent performance was achieved using a free-antibody system with dual calibration. To verify the performance of this method with high reliability, canine C-reactive protein was selected as the immune analyte. Interestingly, our method efficiently mitigated the hook effect with outstanding performance in a one-step reaction without blood dilution or reaction mixing. The detection range of the target can be effectively controlled using free antibodies. Therefore, our CNP-based immunodiagnosis method may advance the commercialization of point-of-care immune biosensors.

A real-time self-powered biosensor is designed for ultrasensitive detection of microRNA-21 based on electrochemical energy device capacitor and target-induced recycling double amplification strategy, which greatly improves the output signal by converting a small number of targets into two glucose oxidase labeled output strand DNAs, and the squeezed-out output strand is recycled by the cathode to fix more signal [Ru(NH3)6]3+ to further improve the detection signal. A digital multimeter (DMM) is connected to computer for real-time displaying the output signal of the self-powered biosensing system, which improves the accuracy of the sensing platform. The sensitivity of the proposed biosensor is 116.15 μA/pM for target microRNA-21, which is 32.26 times higher than that of pure EBFC (3.6 μA/pM). The target concentration is proportional to the open-circuit voltage value in a wide linear range of 0.1-10000 fM with a low detection limit of 0.04 fM (S/N = 3). The method shows high sensitivity and excellent selectivity, and can be applied to detect tumor marker microRNA-21 in biological matrix.

Coreactant electrochemiluminescence (ECL) is one of the most popular pathways in commercial analysis, which can provide simplicity and convenience for getting intense ECL emission. However, the low efficiency of intermolecular electron transfer could weaken ECL intensity. In this work, we developed an enhanced ECL strategy through in situ coordination interactions between metal-organic framework emitters and coreactants. First, a metal-organic framework (MOF) emitter was synthesized with 1,1,2,2-tetrakis(4-(pyridin-4-yl)phenyl)ethane (TPPE) as aggregation-induced emission linkers and Zn as nodes. Interestingly, compared to TPPE ligand, the resulted MOF nanoemitters demonstrated 49.5 folds enhancement of ECL emission in the presence of 1,4-diazabicyclo[2.2.2]octane (DABCO) as the coreactant. More significantly, different from the constant ECL intensity using TPrA coreactant, DABCO exhibited time-dependent ECL intensity due to the intrareticular electron transfer through coordination interaction between DABCO and Zn2+, which was confirmed by X-ray photoelectron spectroscopy and Fourier transform infrared spectral experiments. The enhanced ECL was then applied to construct a sensitive ECL method to detect dopamine in serum samples. The coordination interaction between emitters and coreactants not only provides a universal way to enhance ECL, but also expands the applications of coreactant ECL system in convenience route.

In this work, a facile impedance biosensor was constructed for sensitive assaying of miRNA-10b based on the Cu2+ modified NH2-metal organic frameworks (NMOF@Cu2+) coupling with a three-dimensional (3D) DNA walker signal amplification strategy. Specifically, abundant Cu2+ can adhere to the MOF via the coordination reaction between NH2 and Cu2+, which can be applied as a skeleton to produce CuFe Prussian blue analogue@NMOF (CuFe PBA@NMOF) just in time. Meanwhile, the carboxyl group, which is rich in the organic ligands of the NMOF, can be used to assemble DNA strands (complementary strand, CS) (CS-NMOF@Cu2+) for biorecognition reaction. With the introduction of the target, a 3D DNA walker was triggered to shear out large amounts of assistant strands (AS), which were then anchored on the surface of GCE. Afterward, CS-NMOF@Cu2+ can be assembled on the GCE by hybridization with AS. Eventually, abundant CuFe PBA@NMOF were generated in situ on the electrode with the help of K₃[Fe(CN)6], which can catalyze the 4-chloro-1-naphthol (4-CN) precipitation without H2O2, thereby increasing the resistance of the platform. Under the optimal conditions, the EIS biosensor presents reliable analytical performance in a wide linear range from 0.8 pM to 250 pM with a low detection limit of 0.5 pM.

False detection of SARS-CoV-2 is detrimental to epidemic prevention and control. The scalar nature of the detected signal and the imperfect target recognition property of developed methods are the root causes of generating false signals. Here, we reported a collaborative system of CRISPR-Cas13a coupling with the stabilized graphene field-effect transistor, providing high-intensity vector signals for detecting SARS-CoV-2. In this collaborative system, SARS-CoV-2 RNA generates a "big subtraction" signal with a right-shifted feature, whereas any untargets cause the left-shifted characteristic signal. Thus, the false detection of SARS-CoV-2 is eliminated. High sensitivity with 0.15 copies/μL was obtained. In addition, the wide concerned instability of the graphene field-effect transistor for biosensing in solution environment was solved by the hydrophobic treatment to its substrate, which should be a milestone in advancing it's engineering application. This collaborative system characterized by the high-intensity vector signal and amazing stability significantly advances the accurate SARS-CoV-2 detection from the aspect of signal nature.

The level of hearing restoration in patients with severe to profound sensorineural hearing loss by means of cochlear implants (CIs) has drastically risen since the introduction of these neuroprosthetics. The proposed CI integrated with polymer optical fiber Bragg gratings (POFBGs) enables real-time evaluation of insertion forces and trajectory determination during implantation irrespective of the speed of insertion, as well as provides high signal quality, low stiffness levels, minimum induced stress even under forces of high magnitudes and exhibits significant reduction of the risk of fiber breakage inside the constricted cochlear geometry. As such, the proposed device opens new avenues towards atraumatic cochlear implantations and provides a direct route for the next generation of CIs with intraoperative insertion force assessment and path planning capacity crucial for surgical navigation. Hence, adaptation of this technology to clinical reality holds promising prospects for the hearing impaired.

The localization of isothermal amplification systems has elicited extensive attention due to the enhanced reaction kinetics when detecting ultra-trace small-molecule nucleic acids. Therefore, the seek for an appropriate localization cargo of spatially confined reactions is urgent. Herein, we have developed a novel approach to localize the catalytic hairpin assembly (CHA) system into the DNA tile self-assembly nanostructure. Thanks to the precise programming and robust probe loading capacity, this strategy achieved a 2.3 × 105-fold higher local reaction concentration than a classical CHA system with enhanced reaction kinetics in theory. From the experimental results, this strategy could reach the reaction plateau faster and get access to a magnified effect of 1.57-6.99 times higher in the linear range of microRNA (miRNA) than the simple CHA system. Meanwhile, this strategy satisfied the demand for the one-step detection of miRNA in cell lysates at room temperature with good sensitivity and specificity. These features indicated its excellent potential for ultra-trace molecule detection in clinical diagnosis and provided new insights into the field of bioassays based on DNA tile self-assembly nanotechnology.

Various metabolic diseases are associated with the accumulation of specific amino acids due to abnormal metabolic pathways, and thus can be diagnosed by measuring the level of amino acids in body fluids. However, present methods for amino acid analysis are not readily accessible because they require a complex experimental setup, expensive equipment, and a long processing time. Here, we present a dual sensing microfluidic device that enables fast, portable, and quantitative analysis of target amino acids, harnessing the biological mechanism of protein synthesis. In this device, the working principle of a finger-actuated pumping unit is applied, and the microchannels are designed to perform cell-free synthesis of a reporter protein in response to the target amino acids in the assay samples. Multiple steps required for the translational assay are controlled by the simple operation of two pushbuttons on the device. It is demonstrated that the developed microfluidic device provides precise quantification of two amino acids (methionine and phenylalanine) within 30 min at room temperature. We expect that the application of the presented device can be readily extended to the point-of-care testing of other metabolic compounds.

Exosomal microRNAs (miRNAs) are emerging as attractive non-invasive and reliable biomarkers for disease diagnosis. In situ exosomal miRNA detection can avoid laborious and time-consuming exosome lysis, RNA extraction and effectively improve the accuracy. However, in situ exosomal miRNA detection is hampered by the low abundance of the targets and low permeability of the probes. Herein, an in situ exosomal miRNA sensing biochip based on multi-branched localized catalytic hairpin assembly (MLCHA) and photonic crystals (PCs) was proposed. The MLCHA probes could penetrate into the exosomes nondestructively due to its rigidity and generate amplified fluorescence signal upon recognizing the target miRNA. And then, the fluorescence signal was further enhanced by PCs to improve the sensitivity. The developed biosensor can not only detect exosomal miRNA in a concentration-dependent manner but also distinguish samples from cancer state and healthy state, which is potential for non-invasive clinical diagnostics.

The devastating effects of global climate change on crop production and exponential population growth pose a major challenge to agricultural yields. To cope with this problem, crop performance monitoring is becoming increasingly necessary. In this scenario, the use of sensors and biosensors capable of detecting changes in plant fitness and predicting the evolution of their morphology and physiology has proven to be a useful strategy to increase crop yields. Flexible sensors and nanomaterials have inspired the emerging fields of wearable and on-plant portable devices that provide continuous and accurate long-term sensing of morphological, physiological, biochemical, and environmental parameters. This review provides an overview of novel plant sensing technologies by discussing wearable and integrated devices proposed for engineering plant and monitoring its morphological traits and physiological processes, as well as plant-environment interactions. For each application scenario, the state-of-the-art sensing solutions are grouped according to the plant organ on which they have been installed highlighting their main technological advantages and features. Finally, future opportunities, challenges and perspectives are discussed. We anticipate that the application of this technology in agriculture will provide more accurate measurements for farmers and plant scientists with the ability to track crop performance in real time. All of this information will be essential to enable rapid optimization of plants development through tailored treatments that improve overall plant health even under stressful conditions, with the ultimate goal of increasing crop productivity in a more sustainable manner.

Designing photovoltaic materials with good photoelectric activity is the crucial to boost the sensitivity of photoelectrochemical (PEC) biosensors. To meet this concern, a Schottky-functionalized direct Z-scheme heterojunction photovoltaic material was proposed by electrodeposition of gold nanoparticles on two kinds of bismuth oxyhalide composites surface (bismuth oxybromide and bismuth oxyiodide with different but matched band gaps) (depAu/BiOI/BiOBr). Specifically, synergistic effect was achieved through the direct Z-scheme heterojunction formed by BiOBr and BiOI as well as the gold Schottky junction, resulting in the enhanced light harvest and photoelectric conversion efficiency. Meanwhile, combined with sandwich immunotechnology, a "signal-off" PEC biosensor was fabricated for highly sensitive detection of carcinoembryonic antigen (CEA). In which, using depAu/BiOI/BiOBr modified glassy carbon electrodes both as the photoactive sensing interface and capture antibody loading matrix, polyethyleneimine copper complex encapsulated gold nanoclusters labeled detection antibody (Ab2-Au@PEI-Cu) as the quencher, the photocurrent decreased with the increasing target CEA introduced by sandwich immune reaction. The proposed smart PEC immunoassay platform exhibited a wide detection range (1.0 fg/mL-2.0 ng/mL) and a detection limit as low as 0.11 fg/mL with favorable selectivity and stability. In addition, this PEC sensing strategy can be easily extended for other tumor marker analysis, which offers a new perspective of using multiple bismuth oxyhalide as photoactive materials for early diseases diagnosis.

Convenient, ultrasensitive, and accurate detection of rare variants is essential for early cancer diagnosis and precision medicine, however, despite years of efforts, tools that have all these qualities remain elusive. Here, we developed a one-step CRISPR/Cas12a-based digital diagnostic platform for accurately quantifying mutant alleles, referred to as the CRISPR ASsoaciated Mutation Allele Rapid Test (CASMART). The platform accurately quantifies the variant allele frequency of EGFR L858R within 1 h at 42 °C and can detect mutant targets as low as 0.3 copies/μL (0.498 aM) in mock multiplex cfDNA samples. We further investigated the applicability of CASMART using human genomic samples with confirmed EGFR L858R mutations previously measured variant allele frequency by next-generation sequencing. Comparison across platforms revealed equivalent detection performance (Pearson's correlation coefficient, R2 = 0.9208) and high quantification accuracy for mutation allele frequency (intraclass correlation coefficient = 0.959). Our one-step approach enables easy and accurate variant allele frequency measurement of rare mutant alleles without PCR instrumentation, while the assay time was reduced by approximately half compared to the digital PCR with the shortest turnaround. The CASMART is an alternative to conventional single nucleotide polymorphism detection methods with great potential as a next-generation biosensor for rapidly quantifying the variant allele fraction, especially in resource-limited settings.

The tumor microenvironment consists of a multiplicity of cells such as cancer cells, fibroblasts, endothelial cells, and immune cells within the specific parenchyma. It has been indicated that cancer cells can educate other cells within the tumor niche in a paracrine manner by the release of nano-sized extracellular vesicles namely exosomes (Exo), resulting in accelerated tumor mass growth. It is suggested that exosomal cargo with remarkable information can reflect any changes in metabolic and proteomic profiles in parent tumor cells. Therefore, exosomes can be touted as prognostic, diagnostic, and therapeutic elements with specific biomarkers in patients with different tumor types. Despite the advantages, conventional exosome separation and purification protocols are time-consuming and laborious with low abnormal morphology and purity rate. During the last decades, biosensor-based modalities, as emerging instruments, have been used to detect and analyze Exo in biofluids. Due to suitable specificity, sensitivity, and real-time readout, biosensors became promising approaches for the analysis of Exo in in vitro and in vivo settings. The inherent advantages and superiority of electrochemical biosensors in the determination of tumor grade based on exosomal cargo and profile were also debated. Present and future challenges were also discussed related to the application of electrochemical biosensors in the clinical setting. In this review, the early detection of several cancer types associated with ovaries, breast, brain, colon, lungs, T and B lymphocytes, liver and rare types of cancers were debated in association with released exosomes.

Electronic textiles are fundamentally changing the way we live. However, the inability to effectively recycle them is a considerable burden to the environment. In this study, we developed a cotton fiber-based piezoresistive textile (CF p-textile) for biomonitoring which is biocompatible, biodegradable, and environmentally friendly. These CF p-textiles were fabricated using a scalable dip-coating method to adhere MXene flakes to porous cotton cellulose fibers. The adhesion is made stronger by strong hydrogen bonding between MXene flakes and hierarchically porous cotton cellulose fibers. This cotton-fiber system provides a high sensitivity of 17.73 kPa-1 in a wide pressure range (100 Pa-30 kPa), a 2 Pa subtle pressure detection limit, fast response/recovery time (80/40 ms), and good cycle stability (over 5, 000 cycles). With its compelling sensing performance, the CF p-textile can detect various human biomechanical activities, including pulsation, muscle movement, and swallowing, while still being comfortable to wear. Moreover, the cotton cellulose is decomposed into low-molecular weight cellulose or glucose as a result of the 1,4-glycosidic bond breakage when exposed to acid or during natural degradation, which allows the electronic textile to be biodegradable. This work offers an ecologically-benign, cost-effective and facile approach to fabricating high-performance wearable bioelectronics.

Neutrophil recruitment to the inflamed endothelium is a multistep process and is of utmost importance in the development of the hallmark vaso-occlusive crisis in sickle cell disease (SCD). However, there lacks a standardized, clinically feasible approach for assessing neutrophil recruitment to the inflamed endothelium for individualized risk stratification and therapeutic response prediction in SCD. Here, we describe a microfluidic device functionalized with E-selectin, a critical endothelial receptor for the neutrophil recruitment process, as a strategy to assess neutrophil binding under physiologic flow in normoxia and clinically relevant hypoxia in SCD. We show that hypoxia significantly enhances neutrophil binding to E-selectin and promotes the formation of neutrophil-platelet aggregates. Moreover, we identified two distinct patient populations: a more severe clinical phenotype with elevated lactate dehydrogenase levels and absolute reticulocyte counts but lowered fetal hemoglobin levels associated with constitutively less neutrophil binding to E-selectin. Mechanistically, we demonstrate that the extent of neutrophil activation correlates with membrane L-selectin shedding, resulting in the loss of ligand interaction sites with E-selectin. We also show that inhibition of E-selectin significantly reduces leukocyte recruitment to activated endothelial cells. Our findings add mechanistic insight into neutrophil-endothelial interactions under hypoxia and provide a clinically feasible means for assessing neutrophil binding to E-selectin using clinical whole blood samples, which can help guide therapeutic decisions for SCD patients.

Electrocheluminescence analysis amplified by coreaction accelerators has experienced breakthrough in ultrasensitive detection of biomarkers. Herein, a highly efficient coreaction accelerator, two-dimensional layered MoSe2 nanosheets loaded with gold nanoparticles (AuNPs@MoSe2 heterostructure), is proposed to enhance the ECL efficiency of Ru(bpy)32+/tripropylamine (TPrA) system. The presence of AuNPs avoids the aggregation of MoSe2 nanosheets, and improves the electrical conductivity of modified surface. The AuNPs@MoSe2 modified electrode also provides a large area for loading of abundant capture probe. MoSe2 as an electroactive substrate can remarkably accelerate the generation of TPrA•+ radicals to react with electrooxidized Ru(bpy)32+, which achieves about 3.4-fold stronger ECL intensity. Thus, an enhanced ECL immunoassay method can be achieved after Ru(bpy)32+-doped silica nanoparticle labeled antibody (Ab2-Ru@SiO2) is captured to the modified electrode via immunological recognition. Using methylated DNA as a target, the immunosensor was prepared by binding capture DNA on AuNPs@MoSe2 modified electrode to successively capture the target, anti-5-methylcytosine antibody (anti-5mC) and Ab2-Ru@SiO2. The proposed strategy could detect 0.26 fM 5 mC (3σ) with a detectable concentration range of 1.0 fM - 10 nM at methylated DNA. This immunosensor showed excellent selectivity, good stability and reproducibility, and acceptable recovery, indicating the broad prospects of the novel coreaction accelerator in clinical diagnosis.

Whole-cell biosensors based on transcriptional regulators are powerful tools for rapid measurement, high-throughput screening, dynamic metabolic regulation, etc. To optimize the biosensing performance of transcriptional regulator, its effector-binding domain is commonly engineered. However, this strategy is encumbered by the limitation of diversifying such a large domain and the risk of affecting effector specificity. Molecular dynamics simulation of effector binding of LysG (an LysR-type transcriptional regulator, LTTR) suggests the crucial role of the short linker helix (LH) connecting effector- and DNA-binding domains in protein conformational change. Directed evolution of LH efficiently produced LysG variants with extended operational range and unaltered effector specificity. The whole-cell biosensor based on the best LysGE58V variant outperformed the wild-type LysG in enzyme high-throughput screening and dynamic regulation of l-lysine biosynthetic pathway. LH mutations are suggested to affect DNA binding and facilitate transcriptional activation upon effector binding. LH engineering was also successfully applied to optimize another LTTR BenM for biosensing. Since LTTRs represent the largest family of prokaryotic transcriptional regulators with highly conserved structures, LH engineering is an efficient and universal strategy for development and optimization of whole-cell biosensors.

Neurotransmitter serotonin (5-HT) is involved in various physiological and pathological processes. Therefore, its highly sensitive and selective detection in human serum is of great significance for early diagnosis of disease. In this work, employing iron phthalocyanine as Fe source, ultrafine Fe3O4 nanoparticles anchored on carbon spheres (Fe3O4/CSs) have been prepared, which exhibits an excellent electrochemical sensing performance toward 5-HT. With carbonecous spheres turned into conductive carbon spheres under the heat treatment in N2 atmosphere, iron phthalocyanine absorbed on their surfaces are simultaneously pyrolysised and oxidized, and finally transformed into ultrafine Fe3O4 nanoparticles. Electrochemical results demonstrate a high sensitivity (5.503 μA μM-1) and a low detection limit (4 nM) toward 5-HT for as-prepared Fe3O4/CSs. In combination with the morphology and physicochemical property of Fe3O4/CSs, the enhanced sensing mechanism toward 5-HT is disscussed. In addition, the developed electrochemical sensor also displays a good sensing stability and an anti-interferent ability. Further applied in real human serum samples, a satisfactory recovery rate is achieved. Promisingly, the developed electrochemical sensor can be employed for the determination of 5-HT in actual samples.

Herein, a novel dual-channel electrochemical immunosensor was fabricated via vertical growth of AuPt-decorated boron-doped graphene (AuPt-BG) nanosheets as a signal amplification platform to detect cancer antigen 153 (CA153). Highly open, porous AuPt-BG films were synthesized using one-step electron-assisted hot-filament chemical vapor deposition. The Au-Pt alloy nanoparticles were dispersed on BG nanosheets to improve their biocompatibility, and antibodies (Ab) were directly bonded to the AuPt-BG electrode. The architectures enlarged the loading of CA153Ab and efficiently catalyzed the Fe(CN)63-/4- reaction, ultimately amplifying the signals. This novel strategy allows the simultaneous detection of CA153 in the oxidation and reduction channels, improving the reliability of the detection results. The AuPt-BG-based immunosensor exhibited a lower detection limit (0.0012 mU mL-1, S/N = 3) and wider linear range (0.1-4 × 104 mU mL-1) along with improved reproducibility, selectivity, and stability for the assay of CA153. Owing to the high process controllability of AuPt-BG films, a large-area electrode for in-vitro analyses and a flexible microelectrode for in-vivo analyses were prepared, which confirmed that the AuPt-BG-based sensor is an ideal CA153 detection platform for clinical diagnosis and practical applications.

In situ visualization for the diagnosis of diabetic syndrome and visual monitoring the response to drug treatment is a challenge. Herein, we designed and prepared an autocatalytically-activatable hydrogen peroxide photoacoustic (PA) sensor. We first prepared the FeMoOx nanoparticle with catalase activity, then combined it to 2,2'-azino-bis(3-ethylbenzothi-azoline-6-sulfonic acid) (ABTS) and distearoylphos-phoethanola-mine-polyethylene-glycol (DSPE-PEG) to construct a autocatalytically-activatable PA sensor (FeMoOx@ABTS@DSPE-PEG). In its presence, ABTS can be converted into oxidized ABTS·+ by H2O2. ABTS·+ exhibits strong light absorption in the near-infrared region, and can serve as an ideal contrast agent for PA imaging. H2O2 as a biomarker of oxidative stress response is closely related to the occurrence and development of diabetes mellitus and its complications. Therefore, FeMoOx@ABTS@DSPE-PEG was used as a PA sensor of H2O2 for visual monitoring of the progression of diabetes-induced liver injury and metformin-mediated treatment of diabetes. The autocatalytically-activatable PA sensor developed in this study provides a promising platform for in situ visual diagnosis of diabetes and its syndrome and monitoring the response to therapy.

Herein, we develop a CRISPR/Cas12a-based magnetic relaxation switching (C-MRS) biosensor for ultrasensitive and nucleic acid amplification-free detection of methicillin-resistant Staphylococcus aureus (MRSA) in food. In this biosensor, mecA gene in MRSA was recognized by CRISPR-RNA, which will activate the trans-cleavage activity of Cas12a and release the fastened alkaline phosphatase (ALP) on the particle. The freed ALP can then use to hydrolyze substrate to produce ascorbic acid that trigger the click reaction between magnetic probe. The transverse relaxation time of the unbound magnetic probe can be measured for signal readout. By incorporating collateral activity of CRISPR/Cas12a, on-particle rolling circle amplification, and ALP-triggered click chemistry into background-free MRS, as low as 16 CFU/mL MRSA can be detected without any nucleic acid pre-amplification, which avoids carryover contamination, but without compromising sensitivity. Moreover, this C-MRS biosensor could distinguish 0.01% target DNA from the single-base mutant. Recovery in eggs, milk and pork ranged from 75% to 112%, 82%-104%, and 81%-91%, respectively, revealing its satisfactory accuracy and applicability in the complex food matrix. The developed C-MRS biosensor fleshes out the CRISPR toolbox for food safety and provides a new approach for the sensitive and accurate detection of foodborne drug-resistant bacteria.

We report an aptasensing platform for the detection of cardiac troponin T (cTnT) in the immediate and early phases of acute myocardial infarction (AMI). High-flow filter paper was used to fabricate a microfluidic paper-based analytical device (μ-PAD), which was further modified with gold-decorated polystyrene microparticles functionalized with a highly specific cTnT aptamer. Herein, cTnT detection is presented in two linear ranges (0.01-0.8 μg/ml and 6.25-50 μg/ml) with an LoD of 3.9X10-4 μg/ml, which is in agreement with reference values determined by the American Heart Association. The proposed platform showed remarkable selectivity against AMI-associated cardiac biomarkers such as TNF-alpha, interleukin-6, cardiac troponin I, and reactive protein-C. This aptasensor is a label-free assay that relies only on smartphone-based image analysis and takes less processing time in comparison with traditional methods like ELISA. Furthermore, it exhibits outstanding stability over 23 days when devices are stored at 4 °C. The reported platform is a stable and cost-effective method for the on-site and user-friendly detection of cTnT in normal saline buffer and diluted human serum.

Characterization of cell populations and identification of distinct subtypes based on surface markers are needed in a variety of applications from basic research and clinical assays to cell manufacturing. Conventional immunophenotyping techniques such as flow cytometry or fluorescence microscopy require immunolabeling of cells, expensive and complex instrumentation, skilled operators, and are therefore incompatible with field deployment and automated cell manufacturing systems. In this work, we introduce an autonomous microchip that can electronically quantify the immunophenotypical composition of a cell suspension. Our microchip identifies different cell subtypes by capturing each in different microfluidic chambers functionalized against the markers of the target populations. All on-chip activity is electronically monitored by an integrated sensor network, which informs an algorithm determining subpopulation fractions from chip-wide immunocapture statistics in real time. Moreover, optimal operational conditions within the chip are enforced through a closed-loop feedback control on the sensor data and the cell flow speed, and hence, the antibody-antigen interaction time is maintained within its optimal range for selective immunocapture. We apply our microchip to analyze a mixture of unlabeled CD4+ and CD8+ T cell sub-populations and then validated the results against flow cytometry measurements. The demonstrated capability to quantitatively analyze immune cells with no labels has the potential to enable not only autonomous biochip-based immunoassays for remote testing but also cell manufacturing bioreactors with built-in, adaptive quality control.

Antibody phage display, aimed at preparing antibodies to defined antigens, is a useful replacement for hybridoma technology. The phage system replaces all work stages that follow animal immunization with simple procedures for manipulating DNA and bacteria. It enables the time needed to generate stable antibody-producing clones to be shortened considerably, making the process noticeably cheaper. Antibodies prepared by phage display undergo several affinity selection steps and can be used as selective receptors in biosensors. This article briefly describes the techniques used in the making of phage antibodies to various antigens. The possibilities and prospects are discussed of using phage antibodies as selective agents in analytical systems, including biosensors.

Developing rapid and non-invasive diagnostics for Helicobacter pylori (HP) is imperative to prevent associated diseases such as stomach gastritis, ulcers, and cancers. Owing to HP strain heterogeneity, not all HP-infected individuals incur side effects. Cytotoxin-associated gene A (CagA), and vacuolating cytotoxin A (VacA) genes predominantly drive HP pathogenicity. Therefore, diagnosing CagA and VacA genotypes could alert active infection and decide suitable therapeutics. We report an enhanced LbCas12a trans-cleavage activity with extended reporters and reductants (CEXTRAR) for early detection of HP. We demonstrate that extended ssDNA reporter acts as an excellent signal amplifier, making it a potential alternative substrate for LbCas12a collateral activity. Through a systematic investigation of various buffer components, we demonstrate that reductants improve LbCas12a trans-cleavage activity. Overall, our novel reporter and optimal buffer increased the trans-cleavage activity to an order of 16-fold, achieving picomolar sensitivity (171 pM) without target pre-amplification. Integrated with loop-mediated isothermal amplification (LAMP), CEXTRAR successfully attained attomolar sensitivity for HP detection using real-time fluorescence (43 and 96 aM), in-tube fluorescence readouts (430 and 960 aM), and lateral flow (4.3 and 9.6 aM) for CagA and VacA, respectively. We also demonstrate a rapid 2-min Triton X-100 lysis for clinical sample analysis, which could provide clinicians with actionable information for rapid diagnosis. CEXTRAR could potentially spot the 13C urea breath test false-negatives. For the first time, our study unveils an experimental outlook to manipulate reporters and reconsider precise cysteine substitution via protein engineering for Cas variants with enhanced catalytic activities for use in diagnostics and genetic engineering.

Fat mass and obesity-associated protein (FTO) regulating the N6-methyladenine (m6A, the most pervasive epigenetic modification) levels within the nucleus has been identified as a potential biomarker for cancer diagnosis and prognosis. However, current methods for FTO detection are complicated or/and not sensitive enough for practical application. Herein, we propose a colorimetric biosensor for detecting FTO based on a delicate design of m6A demethylation-activated DNAzyme. Specifically, an m6A-blocked DNAzyme is constructed as a switch of the biosensor that can be turned on by target FTO. The decreased thermal stability resulting from substrate cleavage leads to a DNAzyme recycling to produce multiple primers. Then the rolling circle amplification (RCA) reactions can be initiated to generate G-quadruplex-DNAzymes catalyzing 2,2-azino-bis-(3-ethylben-zthiazoline-6-sulfonic acid (ABTS) oxidation which can be readily observed by the naked eye. Quantitative detection can also be achieved with a limit of detection (LOD) down to 69.9 fM, exhibiting higher sensitivity than previous reports. Therefore, this biosensor opens a simple and sensitive way to achieve visual assay of FTO via triple signal amplification. In addition, our biosensor has been successfully applied to FTO detection in clinical samples, which shows great potential in clinical molecular diagnostics.

Cancer spheroids, which mimic distinct cell-to-cell and cell-extracellular matrix interactions of solid tumors in vitro, have emerged as a promising tumor model for drug screening. However, owing to the unique characteristics of spheroids composed of three-dimensionally densely-packed cells, the precise characterizations of cell viability and function with conventional colorimetric assays are challenging. Herein, we report gold nanostructure-integrated conductive microwell arrays (GONIMA) that enable both highly efficient uniform cancer spheroid formation and precise electrochemical detection of cell viability. A nanostructured gold on indium tin oxide (ITO) substrate facilitated the initial cell aggregation and further 3D cell growth, while the non-cytophilic polymer microwell arrays restricted the size and shape of the spheroids. As a result, approximately 150 human glioblastoma spheroids were formed on a chip area of 1.13 cm2 with an average diameter of 224 μm and a size variation of only 5% (±11.36 μm). The high uniformity of cancer spheroids contributed to the stability of electrical signals measuring cell viability. Using the fabricated GONIMA, the effects of a representative chemotherapeutic agent, hydroxyurea, on the glioblastoma spheroids were precisely monitored under conditions of varying drug concentrations (0-0.3 mg/mL) and incubation times (24-48 h). Therefore, we conclude that the newly developed platform is highly useful for rapid and precise in vitro drug screening, as well as for the pharmacokinetic analyses of specific drugs using 3D cellular cancer models.

Here, a novel and portable machine learning-assisted smartphone-based visual molecularly imprinted ratiometric electrochemiluminescence (MIRECL) sensing platform was constructed for highly selective sensitive detection of 2,4-Dichlorophenoxyacetic acid (2,4-D) for the first time. Te doped CdS-coated Mn3O4 (Te-CdS@Mn3O4) with catalase-like activity served as cathode-emitter, while luminol as anode luminophore accompanied H2O2 as co-reactant, and Te-CdS@Mn3O4 decorated molecularly imprinted polymers (MIPs) as recognition unit, respectively. Molecular models were constructed and MIP band and binding energies were calculated to elucidate the luminescence mechanism and select the best functional monomers. The peroxidase activity and the large specific surface area of Mn3O4 and the electrochemical effect can significantly improve the ECL intensity and analytical sensitivity of Te-CdS@Mn3O4. 2,4-D-MIPs were fabricated by in-situ electrochemical polymerization, and the rebinding of 2,4-D inhibits the binding of H2O2 to the anode emitter, and with the increase of the cathode impedance, the ECL response of Te-CdS@Mn3O4 decreases significantly. However, the blocked reaction of luminol on the anode surface also reduces the ECL response. Thus, a double-reduced MIRECL sensing system was designed and exhibited remarkable performance in sensitivity and selectivity due to the specific recognition of MIPs and the inherent ratio correction effect. Wider linear range in the range of 1 nM-100 μM with a detection limit of 0.63 nM for 2,4-D detection. Interestingly, a portable and visual smartphone-based MIRECL analysis system was established based on the capture of luminescence images by smartphones, classification and recognition by convolutional neural networks, and color analysis by self-developed software. Therefore, the developed MIRECL sensor is suitable for integration with portable devices for intelligent, convenient, and fast detection of 2,4-D in real samples.

P-glycoprotein (P-gp), a transmembrane glycoprotein widely expressed on the surface of various cells, is highly associated with multidrug resistance (MDR) that heralds the malignant progress of disease after drug treatment. Notably, there have been reported that serum P-gp is a potential marker for assessing the progression of disease resistance. Currently, there are few methods for point-of-care serum P-gp detection. In this study, we proposed a gold nanoparticles/electrochemically reduced graphene oxide@carbon nanotube (AuNPs/ERGO@CNT) modified immunosensor based on a one-step electrochemical co-reduction method. The limit of detection (LOD) of our constructed electrochemical immunosensor for P-gp detection reached 0.13 ng/mL, and the detection results in serum were consistent with ELISA. The developed immunosensor is expected to provide a scientific basis for the clinical application of serum P-gp monitoring and integrated medicine.

Rapid diagnostic tests (RDTs) have shown to be instrumental in healthcare and disease control. However, they have been plagued by many inefficiencies in the laborious empirical development and optimization process for the attainment of clinically relevant sensitivity. While various studies have sought to model paper-based RDTs, most have relied on continuum-based models that are not necessarily applicable to all operation regimes, and have solely focused on predicting the specific interactions between the antigen and binders. It is also unclear how the model predictions may be utilized for optimizing assay performance. Here, we propose a streamlined and simplified model-based framework, only relying on calibration with a minimal experimental dataset, for the acceleration of assay optimization. We show that our models are capable of recapitulating experimental data across different formats and antigen-binder-matrix combinations. By predicting signals due to both specific and background interactions, our facile approach enables the estimation of several pertinent assay performance metrics such as limit-of-detection, sensitivity, signal-to-noise ratio and difference. We believe that our proposed workflow would be a valuable addition to the toolset of any assay developer, regardless of the amount of resources they have in their arsenal, and aid assay optimization at any stage in their assay development process.

Surgical resection is a critical procedure for treatment of solid tumor, which commonly suffers from postoperative local recurrence due to the possibility of positive surgical margin. Although the widely used clinical imaging techniques (CT, MRI, PET, etc.) show beneficial effects in providing a macroscopic view of preoperative tumor position, they are still failing to provide intraoperative real-time imaging navigation during the surgery and need oral or intravenous injection contrast agents with risk of adverse effects. In this work, we present a nano-spray assisted photothermal imaging system for in vitro cells discrimination as well as in vivo visualization of tumor position and border that guides real-time precise tumor resection during surgery (even for tiny tumor less than 3 mm). Herein, the nano-spray were prepared by RGD peptide functionalized polydopamine (PDA-RGD) nanospheres with excellent photothermal conversion efficiency (54.27%), stability and reversibility, which target ανβ3 integrin overexpressed tumor cells. Such PDA-RGD serve as nanothermometers that convert and amplify biological signal to intuitive thermal image signal, depicting the tumor margin in situ. In comparison to conventional imaging techniques, our approach through topical spraying together with portable infrared camera has the characteristics of low cost, convenient, no radiation hazard, real-time intraoperative imaging-guidance and avoiding the adverse effects risk of oral or intravenous contrast agent. This technology provides a new universal tool for potentially assisting surgeons' decision in real-time during surgery and aiding to improved outcome.

MicroRNAs (miRNAs) are very important for the early diagnosis and prognosis of tumors. In this work, we achieved the simultaneous detection of microRNA-155 (miR-155) and microRNA-21 (miR-21) with a dual target recognition probe (DRP) based on the nonlinear hybridization chain reaction (HCR). The multi-branched DNA products, three-dimensional multi-hotspot DNA dendrimers (3DmhD) were used in the amplification of the target miRNAs signal. The DRP is constructed with a core of gold nanocages (AuNCs), modified by nucleic acid probes and labeled with Raman signaling molecules ROX and Cy3. Experiments demonstrated that DRP could activate the multi-branched DNA reaction and generate 3DmhD in the presence of miR-155 and miR-21, which can achieve effective amplification of miR-21 and miR-155. When Surface Enhanced Raman Scattering (SERS) analysis was performed on 3DmhD, the multi-hot spot effect of 3DmhD significantly enhanced the signals of ROX and Cy3, allowing ultra-sensitive detection of miR-21 and miR-155 in vitro. To our delight, DRP also exhibited sensitive specificity and significant signal amplification for intracellular miRNAs. These results revealed that DRP has the potential to screen tumor cells by analyzing the expression levels of intracellular miRNAs.

Bioinspired artificial nanochannels have emerged as promising candidates for developing smart nanofluidic sensors due to their highly controllable size and surface functionality. However, little attention has been paid to the role of the outer surface of the nanochannels in enhancing the detection sensitivity. Herein, an asymmetric nanochannel-based responsive detection platform with ultrathin tannic acid modified mesoporous silica (TA-MS) layer and alumina oxide (AAO) thin film is prepared through super-assembly strategy. The functional TA-MS outer surface layer provides abundant phenolic groups on the nanochannels for ions and molecules transport, which paves the way for the development of heterochannels for label-free, reversible and highly sensitive dopamine (DA) detection based off of cation displacement effect. Notably, by engineering optimal thickness of the TA-MS, the sensing performance can be further improved. After optimization, the linear response ranges for DA detection are 0.001-1 μM, 1-10 μM and 10-200 μM with the detection limit of 0.1 nM. The prepared sensor exhibits stable reversibility after several detection cycles. In addition, this method was successfully applied for DA detection in fetal bovine serum sample. Theoretical calculations further prove the detection mechanism. This work opens a new horizon of using mesoporous materials to construct nanofluidic sensors for ultrasensitive small molecule detection and recognition.

Engineered neural tissues serve as models for studying neurological conditions and drug screening. Besides observing the cellular physiological properties, in situ monitoring of neurochemical concentrations with cellular spatial resolution in such neural tissues can provide additional valuable insights in models of disease and drug efficacy. In this work, we demonstrate the first three-dimensional (3D) tissue cultures with embedded optical dopamine (DA) sensors. We developed an alginate/Pluronic F127 based bio-ink for human dopaminergic brain tissue printing with tetrapodal-shaped-ZnO microparticles (t-ZnO) additive as the DA sensor. DA quenches the autofluorescence of t-ZnO in physiological environments, and the reduction of the fluorescence intensity serves as an indicator of the DA concentration. The neurons that were 3D printed with the t-ZnO showed good viability, and extensive 3D neural networks were formed within one week after printing. The t-ZnO could sense DA in the 3D printed neural network with a detection limit of 0.137 μM. The results are a first step toward integrating tissue engineering with intensiometric biosensing for advanced artificial tissue/organ monitoring.

With the increasing concerns of food safety and environmental protection, it is desirable to develop reliable, effective, and portable sensors for detection of organophosphorus pesticides (OPs). Here, a cascade reaction system integrated with threaded 3D microfluidic paper analytical device (3D μPAD) was firstly developed for background-free and visual detection of OPs in agricultural samples. Butyrylcholinesterase (BChE) hydrolyzed acetylcholine into thiocholine (TCh), which reduced MnO2 nanosheets into Mn2+. With addition of OPs, BChE activity was irreversibly inhibited, and the generation of TCh and the reduction of MnO2 nanosheets were prevented. Then the remaining MnO2 nanosheets oxidized o-phenylenediamine into 2,3-diaminophenazine with yellow-emission fluorescence, which quenched the fluorescence intensity of red-emission carbon dots (RCDs) via inner-filter effect. Based on above mechanism, a ratiometric fluorescent system was established for OPs detection. Threaded 3D μPAD consisted of 4 layers, which allowed to load and/or add reagents to trigger the cascade reaction system for OPs detection. The fluorescent images presented distinguishable color variations from red to yellow with dichlorvos concentrations ranging from 2.5 to 120 μg L-1, and the limit of detection was 1.0 μg L-1. In the practical samples testing, threaded 3D μPAD can eliminate background influence on fluorescent signal for OPs detection. Threaded 3D μPAD integrated with ratiometric sensing platform has merits of accuracy response, facile operation, and background-free detection, which supplies a new alternative approach for on-site pesticide detection.

The effective analysis of the basic structure and functional information of bioparticles are of great significance for the early diagnosis of diseases. The synergism between microfluidics and particle manipulation/detection technologies offers enhanced system integration capability and test accuracy for the detection of various bioparticles. Most microfluidic detection platforms are based on optical strategies such as fluorescence, absorbance, and image recognition. Although optical microfluidic platforms have proven their capabilities in the practical clinical detection of bioparticles, shortcomings such as expensive components and whole bulky devices have limited their practicality in the development of point-of-care testing (POCT) systems to be used in remote and underdeveloped areas. Therefore, there is an urgent need to develop cost-effective non-optical microfluidic platforms for bioparticle detection that can act as alternatives to optical counterparts. In this review, we first briefly summarise passive and active methods for bioparticle manipulation in microfluidics. Then, we survey the latest progress in non-optical microfluidic strategies based on electrical, magnetic, and acoustic techniques for bioparticle detection. Finally, a perspective is offered, clarifying challenges faced by current non-optical platforms in developing practical POCT devices and clinical applications.

Phosphorylation of tau at Ser 396, 404 (p-tau396,404) is the earliest phosphorylation event and a promising biomarker for the early diagnosis of Alzheimer's disease (AD). However, the detection of blood p-tau is challenging because of its low abundance, easy degradation, and complex formation with various blood proteins or cells, often leading to the underestimation of p-tau levels in conventional plasma-based assays. Herein, we developed a colorimetric and surface-enhanced Raman scattering (SERS) dual-mode magnetic immunosensor for highly sensitive, specific, and robust detection of p-tau396,404 in whole blood samples. The detection assay was based on an immunoreaction between p-tau396,404 proteins, wherein antibody-modified superparamagnetic iron oxide nanoparticles act as recognition elements to capture p-tau396,404 in blood, and then horseradish peroxidase- and Raman tags label the corresponding paired antibody as a reporter to provide high signal-to-noise ratios for the immunosensor. This dual-mode immunosensor achieved identified as low as 1.5 pg/mL of p-tau396,404 in the blood in SERS mode and 24 pg/mL in colorimetric mode by the naked eye. More importantly, this immunosensor rapidly and accurately distinguished AD patients from healthy individuals based on blood p-tau396,404 levels, and also had the potential to distinguish AD patients of different severities. Therefore, the dual-mode immunosensor is promising for rapid clinical diagnosis of AD, especially in large-scale AD screening.

Microneedle-based wearable sensors offer an alternative approach to traditional invasive blood-based health monitoring and disease diagnostics techniques. Instead of blood, microneedle-based sensors target the skin interstitial fluid (ISF), in which the biomarker type and concentration profile resemble the one found in the blood. However, unlike blood, interstitial fluid does not have the same pH-buffering capacity causing deviation of pH levels from the physiological range. Information about the skin ISF pH levels can be used as a biomarker for a wide range of pathophysiological conditions and as a marker for the calibration of a wearable sensor. The ISF pH can significantly affect the detection accuracy of other biomarkers as it influences enzyme activity, aptamer affinity, and antibody-antigen interaction. Herein, we report the fabrication of a high-density polymeric microneedle array-based (PMNA) sensing patch and its optimization for the potentiometric transdermal monitoring of pH levels in ISF. The wearable sensor utilizes a polyaniline-coated PMNA having a density of ∼10,000 microneedles per cm2, containing individual microneedles with a height of ∼250 μm, and a tip diameter of ∼2 μm. To prevent interference from other body fluids like sweat, an insulating layer is deposited at the base of the PMNA. The wearable pH sensor operates from pH 4.0 to 8.6 with a sensitivity of 62.9 mV per pH unit and an accuracy of ±0.036 pH units. Furthermore, testing on a mouse demonstrates the ability of the PMNA to provide a real-time reading of the transdermal pH values. This microneedle-based system will significantly contribute to advancing transdermal wearable sensors technology, simplifying the fabrication process, and improving the cost-effectiveness of such devices.

Breath acetone (BrAC) detection presents a promising scheme for noninvasive monitoring of metabolic health due to its close correlation to diets and exercise-regulated lipolysis. Herein, we report a Ti3C2Tx MXene-based wireless facemask for on-body BrAC detection and real-time tracking of lipid metabolism, where Ti3C2Tx MXene serves as a versatile nanoplatform for not only acetone detection but also breath interference filtration. The incorporation of in situ grown TiO2 and short peptides with Ti3C2Tx MXene further improves the acetone sensitivity and selectivity, while TiO2-MXene interfaces facilitate light-assisted response calibration. To further realize wearable breath monitoring, a miniaturized flexible detection tag has been integrated with a commercially available facemask, which enables facile BrAC detection and wireless data transmission. Through the hierarchically designed filtration-detection-calibration-transmission system, we realize BrAC detection down to 0.31 ppm (part per million) in breath. On-body breath tests validate the facemask in dynamically monitoring of lipid metabolism, which could guide dieter, athletes, and fitness enthusiasts to arrange diets and exercise activities. The proposed wearable platform opens up new possibility toward the practice of breath analysis as well as daily lipid metabolic management.

Inspired by an adaptive immune system, we have developed a bioelectronic sensing platform which relies on nanovesicles for a signal amplification and can be easily adapted for the detection of new food allergens. In this work, nanovesicles with anti-immunoglobulin E (anti-IgE) antibody receptors were extracted from immune cells and immobilized on a carbon nanotube-based transistor to build a highly sensitive and selective biosensing platform. Our sensor could detect peanut allergen, arachis hypogaea 2 (Ara h 2), down to 0.1 fM and selectively discriminate target allergens in real food samples such as peanut and egg white. As a proof of concept, we demonstrated the detection of different target molecules using the same nanovesicles linked with different antibodies. Our sensor platform was also utilized to quantitatively evaluate the effect of allergy drug such as cromolyn. In this regard, our strategy can be utilized for basic research and versatile applications in food and pharmacological industries.

Preclinical investigation of drug-induced cardiotoxicity is of importance for drug development. To evaluate such cardiotoxicity, in vitro high-throughput interdigitated electrode-based recording of cardiomyocytes mechanical beating is widely used. To automatically analyze the features from the beating signals for drug-induced cardiotoxicity assessment, artificial neural network analysis is conventionally employed and signals are segmented into cycles and feature points are located in the cycles. However, signal segmentation and location of feature points for different signal shapes require design of specific algorithms. Consequently, this may lower the efficiency of research and the applications of such algorithms in signals with different morphologies are limited. Here, we present a biosensing system that employs nonlinear dynamic analysis-assisted neural network (NDANN) to avoid the signal segmentation process and directly extract features from beating signal time series. By processing beating time series with fixed time duration to avoid the signal segmentation process, this NDANN-based biosensing system can identify drug-induced cardiotoxicity with accuracy over 0.99. The individual drugs were classified with high accuracies over 0.94 and drug-induced cardiotoxicity levels were accurately predicted. We also evaluated the generalization performance of the NDANN-based biosensing system in assessing drug-induced cardiotoxicity through an independent dataset. This system achieved accuracy of 0.85-0.95 for different drug concentrations in identification of drug-induced cardiotoxicity. This result demonstrates that our NDANN-based biosensing system has the capacity of screening newly developed drugs, which is crucial in practical applications. This NDANN-based biosensing system can work as a new screening platform for drug-induced cardiotoxicity and improve the efficiency of bio-signal processing.

The rapid and efficient detection of deafness gene DNA plays an important role in the clinical diagnosis of deafness diseases. This study demonstrates the ultrasensitive detection of complementary DNA (cDNA) by employing a nanointerface-sensitized fiber optic biosensor. The sensor consists of SMF-TNCF-MMF-SMF (abbreviated as STMS) structure with lateral offset. Besides, it is functionalized with a nanointerface of black phosphorus (BP) to enhance the light-matter interaction and eventually improve the sensing performances. Relying on this nanointerface-sensitized sensor, we successfully realize the in-situ detection of cDNA at concentrations ranging from 1 pM to 1 μM, with a sensitivity of 0.719 nm/lgM. The limit of detection (LOD) is as low as 0.24 pM, which is at least two orders of magnitude lower than those of existing methods. The sensor exhibits the advantages of simple operation, fast response, label-free measurement, excellent repeatability, and high selectivity. Our contribution suggests a convenient approach for deafness gene DNA detection and can be extended for general ultra-low concentration DNA detection applications.

Developing low-cost and high-efficiency oxygen reduction reaction (ORR) catalysts is crucial to the commercial application of microbial fuel cell (MFC). Herein, Fe/Co-decorated nitrogen-rich three-dimensional (3D) layer-stacked porous biochar (Fe/Co-NC-x) have been synthesized from silk gel through secondary carbonization of activated carbons which firstly adsorbed metal ions. The multilayer porous structure of Fe/Co-NC-3 contributes to construction of high specific surface area (576 m2 g-1), large pore volume (1.27 cm3 g-1) and many defect structure (ID/IG = 1.004). As expected, with Fe/Co synergistic effect, Fe/Co-NC-3 exhibits excellent ORR performance through 4e- pathway with good methanol resistance. In addition, the performance of MFC using Fe/Co-NC-3 as air-cathode catalyst is more prominent with higher maximum power density (1059.62 ± 30.00 mW m-2) compared to that using NC (668.19 ± 9.84 mW m-2) and commercial Pt/C catalyst (957.33 ± 10.50 mW m-2). Therefore, Fe/Co-NC-3 should be a prospective catalyst in the practical application of fuel cells and other energy devices.

Real-time tracking of neurotransmitter levels in vivo has been technically challenging due to the low spatiotemporal resolution of current methods. Since the imbalance of cortical excitation/inhibition (E:I) ratios are associated with a variety of neurological disorders, accurate monitoring of excitatory and inhibitory neurotransmitter levels is crucial for investigating the underlying neural mechanisms of these conditions. Specifically, levels of the excitatory neurotransmitter L-glutamate, and the inhibitory neurotransmitter GABA, are assumed to play critical roles in the E:I balance. Therefore, in this work, a flexible electrochemical microsensor is developed for real-time simultaneous detection of L-glutamate and GABA. The flexible polyimide substrate was used for easier handling during implantation and measurement, along with less brain damage. Further, by electrochemically depositing Pt-black nanostructures on the sensor's surface, the active surface area was enhanced for higher sensitivity. This dual neurotransmitter sensor probe was validated under various settings for its performance, including in vitro, ex vivo tests with glutamatergic neuronal cells and in vivo test with anesthetized rats. Additionally, the sensor's performance has been further investigated in terms of longevity and biocompatibility. Overall, our dual L-glutamate:GABA sensor microprobe has its unique features to enable accurate, real-time, and long-term monitoring of the E:I balance in vivo. Thus, this new tool should aid investigations of neural mechanisms of normal brain function and various neurological disorders.

Reactive oxygen species (ROS) and nicotinamide adenine dinucleotide (NADH) are important intracellular redox-active molecules involved in various pathological processes including inflammation, neurodegenerative diseases, and cancer. However, the fast dynamic changes and mutual regulatory kinetic relationship between intracellular ROS and NADH in these biological processes are still hard to simultaneously investigate. A dual-channel nanowire electrode (DC-NWE) integrating two conductive nanowires, one functionalized with platinum nanoparticles and the other with conductive polymer, was nanofabricated for the selective and simultaneous real-time monitoring of intracellular ROS and NADH release by mitochondria in single living MCF-7 tumoral cells stimulated by resveratrol. The production of ROS was observed to occur tenths of a second before the release of NADH, a significant new piece of information suggesting a mechanism of action of resveratrol. Beyond the importance of the specific data gathered in this study, this work established the feasibility of simultaneously monitoring multiple species and analyzing their kinetics relationships over sub-second time scales thanks to dual-channel nanowire electrodes. It is believed that this concept and its associated nanoelectrochemical tools might benefit to a deeper understanding of mutual regulatory relationship between intracellular crucial molecular markers during physiological and pathological processes as well as for evaluating medical treatments.

Taking advantages of the catalytic activity of METTL3/METTL14 protein towards adenine methylation in RNA sequence and the specific digestion activity of MazF protein towards unmethylated RNA sequence containing ACA bases, a novel photoelectrochemical biosensor was constructed for simultaneous detection of RNA methylation, METTL3/METTL14 protein and MazF protein. MoSe2-BiOI nanocomposite was prepared and considered as photoactive material, catalytic hairpin assembly strategy and in situ generation of electron donors catalyzed by polyaspartic acid-loaded alkaline phosphatase technique were employed as signal amplification. Under the optimum conditions, the detection ranges of methylated RNA, METTL3/METTL14 protein and MazF protein were 0.001-50 nM, 0.001-25 ng/μL, and 0.001-10 U/mL, respectively. The corresponding detection limits were 0.46 pM, 0.51 pg/μL and 0.42 U/μL with S/N = 3. In addition, the effect of drugs and composite pollutants on the activities of MazF proteins was assessed, proving the applicability of the developed method in the field of drug screening for MazF-related diseases. Moreover, the effects of pollutants on the activity of METTL3/METTL14 were also preliminarily explored, providing new information on pathogenic mechanism of pollutants.

The development of space exploration technologies has positively impacted everyday life on Earth in terms of communication, environmental, social, and economic perspectives. The human body constantly fluctuates during spaceflight, even for a short-term mission. Unfortunately, technology is evolving faster than humans' ability to adapt, and many therapeutics entering clinical trials fail even after being subjected to vigorous in vivo testing due to toxicity and lack of efficacy. Therefore, tissue chips (also mentioned as organ-on-a-chip) with biosensors are being developed to compensate for the lack of relevant models to help improve the drug development process. There has been a push to monitor cell and tissue functions, based on their biological signals and utilize the integration of biosensors into tissue chips in space to monitor and assess cell microenvironment in real-time. With the collaboration between the Center for the Advancement of Science in Space (CASIS), the National Aeronautics and Space Administration (NASA) and other partners, they are providing the opportunities to study the effects of microgravity environment has on the human body. Institutions such as the National Institute of Health (NIH) and National Science Foundation (NSF) are partnering with CASIS and NASA to utilize tissue chips onboard the International Space Station (ISS). This article reviews the endless benefits of space technology, the development of integrated biosensors in tissue chips and their applications to better understand human biology, physiology, and diseases in space and on Earth, followed by future perspectives of tissue chip applications on Earth and in space.

Nucleic acid detection is crucial for monitoring diseases for which rapid, sensitive, and easy-to-deploy diagnostic tools are needed. CRISPR-based technologies can potentially fulfill this need for nucleic acid detection. However, their widespread use has been restricted by the requirement of a protospacer adjacent motif in the target and extensive guide RNA optimization. In this study, we developed FELICX, a technique that can overcome these limitations and provide a useful alternative to existing technologies. FELICX comprises flap endonuclease, Taq ligase and CRISPR-Cas for diagnostics (X) and can be used for detecting nucleic acids and single-nucleotide polymorphisms. This method can be deployed as a point-of-care test, as only two temperatures are needed without thermocycling for its functionality, with the result generated on lateral flow strips. As a proof-of-concept, we showed that up to 0.6 copies/μL of DNA and RNA could be detected by FELICX in 60 min and 90 min, respectively, using simulated samples. Additionally, FELICX could be used to probe any base pair, unlike other CRISPR-based technologies. Finally, we demonstrated the versatility of FELICX by employing it for virus detection in infected human cells, the identification of antibiotic-resistant bacteria, and cancer diagnostics using simulated samples. Based on its unique advantages, we envision the use of FELICX as a next-generation CRISPR-based technology in nucleic acid diagnostics.

The organized self-assembly of conductive biological structures holds promise for creating new bioelectronic devices. In particular, Geobacter sulfurreducens type IVa pili have proven to be a versatile material for fabricating protein nanowire-based devices. To scale the production of conductive pili, we designed a strain of Shewanella oneidensis that heterologously expressed abundant, conductive Geobacter pili when grown aerobically in liquid culture. S. oneidensis expressing a cysteine-modified pilin, designed to enhance the capability to bind to gold, generated conductive pili that self-assembled into biohybrid filaments in the presence of gold nanoparticles. Elemental composition analysis confirmed the filament-metal interactions within the structures, which were several orders of magnitude larger than previously described metal:organic filaments. The results demonstrate that the S. oneidensis chassis significantly advances the possibilities for facile conductive protein nanowire design and fabrication.

Fungal infections are a rapidly increasing public health problem due to their high morbidity and mortality rates, especially in populations with compromised immune systems. Rapid and accurate diagnosis of these diseases is, therefore, necessary to improve the prognosis of afflicted patients. Unfortunately, current clinical chemistry practice relies on lengthy culturing methods that are insufficient to meet the fast turnaround requirements. Here we present a cost-effective and robust nucleic acid sensor that can identify the presence of histoplasmosis causing fungal genes, in whole blood or bronchoalveolar lavage (BAL) samples, far earlier than current methods. Our novel assay involves the hybridization of target gene sequences with immobilized nucleic acid probes, allowing direct, label-free detection of Hcp100, CBP1, and M antigen genes through electrochemical analysis. The resultant current is attributed to the presence of fungal targets in the sample solution. The assay provides ultra-sensitive detection of DNA molecules with a limit of detection (LOD) values down to 100 aM, sufficient to meet the clinical diagnostic need. In addition, the turnaround time for the sample to result is less than 90 min compared to the current clinical procedure's turnaround time of 3-4 weeks.

Organic thin-film transistors (TFTs) with an electrochemically functionalized sensing gate are promising platforms for wearable health-monitoring technologies because they are light, flexible, and cheap. Achieving both high sensitivity and low power is highly demanding for portable or wearable devices. In this work, we present flexible printed dual-gate (DG) organic TFTs operating in the subthreshold regime with ultralow power and high sensitivity. The subthreshold operation of the gate-modulated TFT-based sensors not only increases the sensitivity but also reduces the power consumption. The DG configuration has deeper depletion and stronger accumulation, thereby further making the subthreshold slope sharper. We integrate an enzymatic lactate-sensing extended-gate electrode into the printed DG TFT and achieve exceptionally high sensitivity (0.77) and ultralow static power consumption (10 nW). Our sensors are successfully demonstrated in physiological lactate monitoring with human saliva. The accuracy of the DG TFT sensing system is as good as that of a high-cost conventional assay. The developed platform can be readily extended to various materials and technologies for high performance wearable sensing applications.

The process of developing an end-to-end model of a magneto-immunoassay is described, simulating the agglutination effect due to the specific binding of bacteria to paramagnetic particles. After establishing the properties of the dose-specific agglutination through direct imaging, a microfluidic assay was used to demonstrate changes in the magnetophoretic transport dynamics of agglutinated clusters via transient inductive magentometer measurements. End-to-end mathematical modelling is used to establish the physical processes underlying the assay. First, a modified form of Becker-Döring nucleation kinetic equations is used to establish a relationship between analyte dose and average cluster size. Next, Stokes flow equations are used to establish a relationship between cluster size and speed of motion within the fluid chamber. This predicts a cluster-size dynamic profile of concentration of PMPs versus time when the magnetic field is switched between the two actuated magnets. Finally, inductive modelling is carried out to predict the response of the magnetometer circuit in response to the dynamics of magnetic clusters. The predictions of this model are shown to agree well with the results of experiments, and to predict the shape of the dose-response curve.

Urinary tract infections (UTIs) are among the most predominant microbial diseases, leading to substantial healthcare burdens and threatening human well-being. UTIs can become more critical when caused by Pseudomonas aeruginosa, particularly by antimicrobial-resistant types. Thereby a rapid diagnosis and identification of the antimicrobial-resistant P. aeruginosa can support and guide an efficient medication and an effective treatment toward UTIs. Herein, we designed a platform for prompt purification, and effective identification of P. aeruginosa to combat the notorious P. aeruginosa associated UTIs. A peptide (QRKLAAKLT), specifically binding to P. aeruginosa, was grafted onto PEGylated magnetic nanoclusters and enabled a successful capture and enrichment of P. aeruginosa from artificial human urine. Rapid identification of antimicrobial resistance of the enriched P. aeruginosa can be moreover accomplished within 30 min. These functionalized magnetic nanoclusters demonstrate a prominent diagnostic potential to combat P. aeruginosa associated UTIs, which can be extended to other P. aeruginosa involved infections.

In order to achieve rapid and sensitive detection of CYFRA 21-1, a signal-off photoelectrochemical (PEC) immunosensor was devised with NiCo2O4/CdIn2S4/In2S3 heterojunction photoactive materials as sensing platform and ReS2@Au NPs as the secondary antibody labels amplifying signal based on the energy band-matching cascade structure and double suppression effect. NiCo2O4 possessed a faster charge transfer rate due to the abundance of redox electron pairs (Co3+/Co2+ and Ni3+/Ni2+). To further improve the PEC properties of NiCo2O4 under visible light, CdIn2S4 with matching bandgap energy was selected to form heterojunction with NiCo2O4 and sensitized with In2S3. The proposed heterojunctions with well-matched band structure promoted the transfer of photo-generated carriers and were exploited as signal transducers for immobilization of antibodies and recognition of CYFRA 21-1. Furthermore, a novel urchin-like p-type ReS2 semiconductor nanostructure functionalized by Au NPs was firstly used as a nanolabel to quench the signal. On the one hand, the Schottky heterojunction generated by ReS2 and Au NPs could compete with the transducer substrate for both light and electron donors. On the other hand, the large space steric hindrance of ReS2 prevented contact between the matrix and AA. Subsequently, the sensor was sensitive in a wide range of concentrations for CYFRA 21-1 (0.0001-50 ng/mL), and the detection limit was 0.05 pg/mL.

A simple, affordable point of care test (POCT) is necessary for on-site detection of coronavirus disease 2019 (COVID-19). The lateral flow assay (LFA) has great potential for use in POCT mainly because of factors such as low time consumption, low cost, and ease of use. However, it lacks sensitivity and limits of detection (LOD), which are essential for early diagnostics. In this study, we proposed a non-powered preconcentrator (NPP) based on nanoelectrokinetics for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Antigen (Ag) lateral flow assay. The non-powered preconcentrator is composed of glass fiber-based composite paper and ion permselective material, and it can be simply operated by force balancing gravitational, capillary, and depletion-induced forces. The proposed approach helps enrich the SARS-CoV-2 viral nucleocapsid (N) proteins based on a 10-min operation, and it improved the LOD by up to 10-fold. The corresponding virus enrichment, which was evaluated using the reverse-transcriptase polymerase chain reaction (RT-PCR), revealed an improvement in ΔCt values > 3. We successfully demonstrated the enhancement of the NPP-assisted LFA, we extended to applying it to clinical samples. Further, we demonstrated an affordable, easy-to-implement form of LFA by simply designing NPP directly on the LFA buffer tube.

The accurate detection of microRNAs (miRNAs) is essential in the early diagnosis and treatment of cancers. Existing miRNA detection methods represented by nucleic acid amplification (NAA) techniques, such as qRT-PCR, suffer from the small size of miRNAs and lead to limited practicability. CRISPR Cas13a system, another valuable toolbox for nucleic acid detection, relies heavily on the behaviors of accompanying isothermal NAA techniques, which prompts similar deficiencies in miRNA detection. In this study, a dual nucleases-assisted cyclic amplification (DUNCAN) strategy has been established to replace NAA techniques for one-pot detection of miRNAs. The DUNCAN strategy contained an initial reaction based on CRISPR Cas13a for target recognition, and an accompanied cyclic reaction using DNA probes protected by polydopamine nanospheres (PDANSs) for signal amplification and result readout. Exemplified by miR-19b, which has been confirmed to be related to several tumors, the quantitative detection through the DUNCAN strategy was achieved in the dynamic range of 10-106 fM, with a calculated detection limit of 1.27 fM. Besides, the DUNCAN strategy presented well selectivity and anti-interference performance for accurate detection of miR-19b in complex miRNA mixtures, different cell lines and clinical samples compared with qRT-PCR. All these performances demonstrated the promising potential of the DUNCAN strategy in clinical miRNA detection and diagnosis.

Stereochemical analysis is essential for understanding the complex function of biomolecules. Various direct and indirect approaches can be used to explore the allosteric configuration. However, the size, cost, and delicate nature of these systems limit their biomedical usage. Here, we constructed elliptical dichroism (ED) spectrometer for biomedical applications, whose performance is validated by experiment and theoretical simulation (Jones/Mueller calculus and time-dependent density-functional theory). Instead of complicated control of circular polarization, ED spectrometer adopted the absorbance of left- and right-oriented elliptically polarized light. With a simplified design, we demonstrated the potential of ED spectrometry as an alternative for secondary structural analysis of biomolecules, their conformation and chirality. It not only provides a portable, low-cost alternative to the sophisticated instruments currently used for structural analysis of biomolecules but also provides superior translational features: low sample consumption(200 μl), easy operation, and multiple working modes, for noninvasive cancer detection.