Improve Detection Sensitivity Research Articles

Sensitivity and specificity of in vivo COVID-19 screening by detection dogs: Results of the C19-Screendog multicenter study

Trained dogs can recognize the volatile organic compounds contained in biological samples of patients with COVID-19 infection. We assessed the sensitivity and specificity of in vivo SARS-CoV-2 screening by trained dogs.We recruited five dog-handler dyads. In the operant conditioning phase, the dogs were taught to distinguish between positive and negative sweat samples collected from volunteers' underarms in polymeric tubes. The conditioning was validated by tests involving 16 positive and 48 negative samples held or worn in such a way that the samples were invisible to the dog and handler. In the screening phase the dogs were led by their handlers to a drive-through facility for in vivo screening of volunteers who had just received a nasopharyngeal swab from nursing staff. Each volunteer who had already swabbed was subsequently tested by two dogs, whose responses were recorded as positive, negative, or inconclusive. The dogs’ behavior was constantly monitored for attentiveness and wellbeing.All the dogs passed the conditioning phase, their responses showing a sensitivity of 83–100% and a specificity of 94–100%. The in vivo screening phase involved 1251 subjects, of whom 205 had a COVID-19 positive swab and two dogs per each subject to be screened. Screening sensitivity and specificity were respectively 91.6–97.6% and 96.3–100% when only one dog was involved, whereas combined screening by two dogs provided a higher sensitivity. Dog wellbeing was also analyzed: monitoring of stress and fatigue suggested that the screening activity did not adversely impact the dogs’ wellbeing. This work, by screening a large number of subjects, strengthen recent findings that trained dogs can discriminate between COVID-19 infected and healthy human subjects and introduce two novel research aspects: i) assessement of signs of fatigue and stress in dogs during training and testing, and ii) combining screening by two dogs to improve detection sensitivity and specificity.Using some precautions to reduce the risk of infection and spillover, in vivo COVID-19 screening by a dog-handler dyad can be suitable to quickly screen large numbers of people: it is rapid, non-invasive and economical, since it does not involve actual sampling, lab resources or waste management, and is suitable to screen large numbers of people.

Fabrication of trastuzumab conjugated curcumin nanoparticles based impedimetric cytosensor for the cancer cell detection

In the study, trastuzumab-modified curcumin nanoparticles (Anti-Cur NPs) functionalized screen printed electrode (SPE) was constructed for targeted BT-474 cells diagnosis as the cytosensing interface of a novel impedance cytosensor. Cur NPs with multifunctional properties were fabricated by the wet milling-solvent evaporation method to produce biologically compatible NPs without any further operation. Then, trastuzumab, which was selected for increasing nanocurcumin adherence affinity notably for cancer cells, was attached to the surface of Cur NPs. Based on the characterization studies, Anti-Cur NPs functionalized cytosensor displayed suitable surfaces due to hydrophilic, biocompatible, and targeted features where cancer cells could attach efficiently. The offered impedimetric diagnosis depended on the peculiar interaction between trastuzumab functional groups and HER-2 receptors which were highly overexpressed on breast cancer cells (BT-474). The findings revealed that the developed Anti-Cur NPs based platform enhanced the impedimetric signal and improved detection sensitivity for the BT-474 cells was attained in a vast linear range (1.0 × 102 – 8.0 × 104 cells mL−1) with a low detection limit (65 cells mL−1). Thus, the constructed cytosensor enabled impedance based monitoring of the cell behavior regarding the processes of cell attachment in a quantitative aspect with highly reproducible responses. Anti-Cur NP/SPE based cytosensor platform provided sensitive and fast measurement of the low amount of BT-474 cells in early diagnosis with an outstanding performance which was assigned to the well-defined and targeted nanocurcumin on the substrate.

Abstract 1535: A novel ELISA Kit:highly accuracy and sensitivity for GDF-15 detection

Abstract Growth/differentiation factor 15 (GDF-15) is a promising drug target and biomarker for cancer early screening or immunotherapy. Under normal conditions, GDF-15 is expressed in low concentrations in most organs. Increased protein levels are associated with disease states such as tissue hypoxia, inflammation, acute injury and oxidative stress. The development of GDF-15 detection kit can help pharmaceutical companies in the early stage of drug development. It can also facilitate the early screening of related diseases. Hence, we developed a novel GDF-15 detected ELISA kit based on double-antibody sandwich ELISA. Our kit has ultra-high sensitivity and specificity and the concentration of GDF-15 can be detected accurately in complex biological samples such as serum, plasma or urine. First of all, mice were immunized by GDF-15. After detecting the antibody titers in sera of the immunized mice, hybridoma technologies were used to produce GDF-15 monoclonal antibodies. 57 antibodies were analyzed by epitope competition, ELISA binding and KD binding detection. The specificity of pairwise candidates was screened to obtain optimal coating and detecting antibody pair. The detecting antibody was labeled with biotin which can be conjugated with streptavidin labeled HRP. Biotin-streptavidin labeling detection method can achieve signal amplification which offers an improved detection sensitivity. Our GDF-15 detection kit has better property in sensitivity and specificity. The detection range is from 10 pg/mL to 10000 pg/mL. LOD is 5 pg/mL and LOQ is 10 pg/mL. These parameters are better than many commercial test kits. Moreover, the concentration of GDF-15 can be detected accurately in complex clinical samples. We anticipate that this GDF-15 detection kit can be applied not only for companion diagnosis, but also for related drug development. Citation Format: Meng Liang, Feng He, Meng Zhang, Yinsong Ye, Shukun Chen, Jinying Ning, Feng Hao. A novel ELISA Kit:highly accuracy and sensitivity for GDF-15 detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1535.

Ultrasensitive visual detection of the food-borne pathogen via MOF encapsulated enzyme

Various methods have been made to achieve sensitive detection (10 CFU/mL) of Escherichia coli O157:H7 (E. coli) in real samples, however, they are complex, time-consuming, or instrument-dependent. Enzyme-catalyzed reactions are one of the most efficient methods to amplify signals for sensitive detection. ZIF-8 owning stability, porosity, and high specific area are suitable for embedding enzymes which can effectively protect enzyme activity and thus improve detection sensitivity. Herein, a simple visual assay of E. coli with the limits of detection of 1 CFU/mL was developed based on this stable enzyme-catalyzed amplified system. A microbial safety test of milk, orange juice, seawater, cosmetic, and hydrolyzed yeast protein, was successfully performed with the limits of detection of 10 CFU/mL by the naked eye. And this bioassay possessed high selectivity and stability making the developed detection method practically promising.

A Magnetic Shielding-Type PEC Sensor With a Canister Structure and Magnetic Core

The pulsed eddy current (PEC) technique exhibits considerable benefits over conventional eddy current techniques. The structure and dimensional parameters of PEC sensors considerably influence the PEC systems. Conventional PEC sensors are susceptible to background noise of a magnetic excitation field. To strengthen the magnetic gathering ability and improve detection sensitivity, a magnetic shielding shell structure was introduced to PEC sensors. In this study, a novel canister structure and magnetic core-based PEC sensor with magnetic shielding was designed. Finite-element simulation and experimental study were performed subsequently to optimize the design. A 3-D finite-element simulation model was established according to the eddy current detection mechanism and electromagnetic shielding principle. The influence of the PEC sensor structure and size on its detection performance was observed. Next, a 2-D axis-symmetric model was established, and the size of this PEC sensor was optimized by using the orthogonal approach. The results revealed that the detection performance of this PEC sensor could be improved by incorporating an appropriate magnetic shielding mechanism and through the optimization of the sensor size. The research results show that compared with the PEC sensor without magnetic shield, the PEC sensor with magnetic shield developed in this article can obtain higher signal amplitude and can be better applied to surface and subsurface defect detection.

Single-Site SnOCu Pairs with Interfacial Electron Transfer Effect for Enhanced Electrochemical Catalysis and Sensing.

As advanced electrochemical catalysts, single-atom catalysts have made great progress in the field of catalysis and sensing due to their high atomic utilization efficiency and excellent catalytic performance. Herein, stannum-doped copper oxide (CuOSn1 ) nanosheets with single-site SnOCu pairs as active sites are synthesized as electrocatalysts for biological molecule detection. Compared with CuO-based electrochemical sensors, the CuOSn1 -based electrochemical sensors have improved detection sensitivity with a rapid electrochemical response. Theoretical calculation reveals that the single-site SnOCu pairs induced interfacial electronic transfer effect can strengthen hydroxy adsorption and thus reduce the energy barrier of the biological molecule oxidation process. As a concept application, electrochemical detection of dopamine and uric acid molecules is achieved, exhibiting satisfactory sensitivity and selectivity. This work demonstrates the advantages of single-site SnOCu pairs in electrochemical catalysis and sensing, which provides theoretical guidance for understanding the structure-activity relationship for sensitive electrochemical sensing.

Efficient removal of phytochrome using rice straw-derived biochar: Adsorption performance, mechanisms, and practical applications

Rice straw derived biochar was fabricated and applied as a purification agent. The adsorption kinetics, isotherms, and thermodynamics for adsorbates were determined using the biochar. Adsorption kinetics and isotherms were best fitted by the pseudo-second order and Langmuir models. Biochar could effectively remove chlorophyll in 9 different solutions. Biochar was employed as a clean-up reagent for 149 pesticides detection, which revealed that biochar had a higher phytochrome removal capacity than graphitized carbon black and 123 pesticides had satisfactory recovery values. The biochar was prepared into a sample pad by electrospinning and was then used for online sample clean-up in a test strip, and it showed high ability of removing phytochrome and improving detection sensitivity. Thus, biochar could be applied as a purification agent to remove pigmentation, making it a promising candidate not only for sample pretreatment but also in the fields of food, agriculture and environment.

On the Investigation of Frequency Characteristics of a Novel Inductive Debris Sensor.

Lubricants have the ability to reduce frictions, prevent wear, convey metal debris particles and increase the efficiency of heat transfer; therefore, they have been widely used in mechanical systems. To assess the safety and reliability of the machine under operational conditions, the development of inductive debris sensors for the online monitoring of debris particles in lubricants has received more attention from researchers. To achieve a high-precision, high-efficiency sensor for accurate prediction on the degree of wear, the equivalent circuit model of the sensor coil has been established, and its equations discovering the relationship between the induced voltage and excitation frequency have been derived. Furthermore, the influence of excitation frequencies and metal debris on the magnetic flux density has been analyzed throughout the simulations to determine the sensor magnetic field. In order to identify a frequency range suitable for detecting both ferrous and non-ferrous materials with a high level of sensitivity, the analytical analysis and experiments have been conducted to investigate the frequency characteristics of the developed inductive debris sensor prototype and its improved inspection capability. Moreover, the developed inductive debris sensor with the noticeable frequency characteristics has been assessed and its theoretical model has been also validated throughout experimental tests. Results have shown that the detection sensitivity of non-ferrous debris by the developed sensor increases with the excitation frequency in the range of 50 kHz to 250 kHz, while more complex results for the detection of ferrous debris have been observed. The detection sensitivity decreases as the excitation frequency increases from 50 kHz to 300 kHz, and then increases with the excitation frequency from 300 kHz to 370 kHz. This leads to the effective selection of the excitation frequency in the process of inspection. In summary, the investigation into the frequency characteristics of the proposed novel inductive debris sensor has enabled its broad applications and also provided a theoretical basis and valuable insights into the development of inductive debris sensors with improved detection sensitivity.

On-tissue chemical derivatization enables spatiotemporal heterogeneity visualization of oxylipins in esophageal cancer xenograft via ambient mass spectrometry imaging

On-tissue chemical derivatization (OTCD) effectively enhances ionization efficiency of low abundant and poorly ionized functional molecules to improve detection sensitivity and coverage of mass spectrometry imaging (MSI). Combination OTCD and MSI provides a novel strategy for visualizing previously undisclosed metabolic heterogeneity in tumor. Herein, we present a method to visualize heterogeneous metabolism of oxylipins within tumor by coupling OTCD with airflow-assisted desorption electrospray ionization (AFADESI)-MSI. Taking Girard's P as a derivatization reagent, easily ionized hydrazide and quaternary amine groups were introduced into the structure of carbonyl metabolites via condensation reaction. Oxylipins, including 127 fatty aldehydes (FALs) and 71 oxo fatty acids (FAs), were detected and imaged in esophageal cancer xenograft with AFADESI-MSI after OTCD. Then t-distributed stochastic neighbor embedding and random forest were exploited to precisely locate the distribution of oxylipins in heterogeneous tumor tissue. With this method, we surprisingly found almost all FALs and oxo FAs significantly accumulated in the core region of tumor, and exhibited a gradual increase trend in tumor over time. These results reveal spatiotemporal heterogeneity of oxylipins in tumor progression, highlighting the value of OTCD combined with MSI to gain deeper insights into understanding tumor metabolism.

Cysteine-Activatable Near-Infrared Fluorescent Probe for Dual-Channel Tracking Lipid Droplets and Mitochondria in Epilepsy.

Dual-channel fluorescent probes could respond to a specific target and emit different wavelengths of fluorescence before and after the response. Such probes could alleviate the influence caused by the variation of the probe concentration, excitation intensity, and so on. However, for most dual-channel fluorescent probes, the probe and fluorophore faced spectral overlap, which reduced sensitivity and accuracy. Herein, we introduced a cysteine (Cys)-responsive and near-infrared (NIR) emissive AIEgen (named TSQC) with good biocompatibility to dual-channel monitor Cys in mitochondria and lipid droplets (LDs) during cell apoptosis through wash-free fluorescence bio-imaging. TSQC can label mitochondria with bright fluorescence around 750 nm, and after reacting with Cys, the reaction product TSQ could spontaneously target LDs with emissions around 650 nm. Such spatially separated dual-channel fluorescence responses could significantly improve detection sensitivity and accuracy. Furthermore, the Cys-triggered dual-channel fluorescence imaging in LDs and mitochondria during apoptosis induced by UV light exposure, H2O2, or LPS treatment is clearly observed for the first time. Besides, we also report here that TSQC can be used to image subcellular Cys in different cell lines by measuring the fluorescence intensities of different emission channels. In particular, TSQC shows superior utility for the in vivo imaging of apoptosis in acute and chronic epilepsy mice. In brief, the newly designed NIR AIEgen TSQC can respond to Cys and separate two fluorescence signals to mitochondria and LDs, respectively, to study Cys-related apoptosis.

Learning to detect auditory signals in noise: Active top-down selection and stable change in signal representations.

Training can improve detection of auditory signals in noise. This learning could potentially occur through active top-down selection mechanisms or stable changes in signal representations. Here, participants were trained and tested (pretest vs. posttest design) on abilities to detect pure tone signals in noise. Auditory evoked potentials (AEPs) to tones were gathered under dichotic listening conditions where participants attended to nontonal stimuli in the opposite ear. Improvements in detection sensitivity were observable regardless of tested tone frequency. This was true in generalization between 861 Hz and 1058-Hz tones (Experiment 1a), and when testing a frequency range > 1 octave (Experiment 2). Such learning was not apparent without training (Experiment 1b). In contrast to behavior, AEP amplitude increases from pre- to posttest were partially specific to trained tone frequencies, even when selective attention was diverted to the opposite ear of tone presentation. Placed in the context of previous work, results suggest that changes in active top-down selection mechanisms and stable signal representations both play a role in auditory detection learning. The mismatch between AEP and behavioral effects suggests a need to consider how these different learning processes can impact detection performance in the variety of listening scenarios a listener may face. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Determination of Anthraquinone-Tagged Amines Using High-Performance Liquid Chromatography with Online UV Irradiation and Luminol Chemiluminescence Detection

Quinones are frequently used as derivatization reagents in HPLC analysis to improve detection sensitivity. In the present study, a simple, sensitive, and selective chemiluminescence (CL) derivatization strategy for biogenic amines, prior to their HPLC-CL analysis, was developed. The novel CL derivatization strategy was established based on using anthraquinone-2-carbonyl chloride as derivatizing agent for amines and then using the unique property of the quinones' moiety to generate reactive oxygen species (ROS) in response to UV irradiation. Typical amines such as tryptamine and phenethylamine were derivatized with anthraquinone-2-carbonyl chloride and then injected into an HPLC system equipped with an online photoreactor. The anthraquinone-tagged amines are separated and then UV-irradiated when they pass through a photoreactor to generate ROS from the quinone moiety of the derivative. Tryptamine and phenethylamine can be determined by measuring the chemiluminescence intensity produced by the reaction of the generated ROS with luminol. The chemiluminescence disappears when the photoreactor is turned off, suggesting that ROS are no longer generated from the quinone moiety in the absence of UV irradiation. This result indicates that the generation of ROS could be controlled by turning the photoreactor on and off. Under the optimized conditions, the limits of detection for tryptamine and phenethylamine were 124 and 84 nM, respectively. The developed method is successfully applied to determine the concentrations of tryptamine and phenethylamine in wine samples.

Sensitive determination of inosine RNA modification in single cell by chemical derivatization coupled with mass spectrometry analysis

Inosine is a vital RNA modification across three kingdoms of life. It has been demonstrated that inosine plays important roles in modulation of the fate of RNAs. In the current study, we developed a highly sensitive method to determine inosine in a single cell by N-cyclohexyl-N’-β-(4-methylmorpholinium)ethylcarbodiimide p-toluenesulfonate (CMCT) derivatization in combination with mass spectrometry analysis. The results showed that the detection sensitivity of inosine was increased by 556-fold after CMCT derivatization, with the limit of detection (LOD) being 4.5 amol. With the established method, we could detect inosine from 13.0 pg of total RNA of HEK293T cells. Meanwhile, inosine in RNA from a single cell could also be clearly detected due to the improved detection sensitivity. Moreover, we found the level of inosine in RNA of sleep-deprived mice was significantly increased compared to the control mice, indicating that inosine is associated with sleep behavior and might be a potential indicator of sleep disorder. Taken together, the chemical derivatization coupled with mass spectrometry analysis offers a valuable tool in determination of endogenous RNA modifications in a single cell, which should benefit the functional study of RNA modification in rare clinical samples.

Fabrication of CuInZnS/ZnS Quantum Dot Microbeads by a Two-Step Approach of Emulsification-Solvent Evaporation and Surfactant Substitution and Its Application for Quantitative Detection.

CuInS2 quantum dots (CIS QDs) are considered to be promising alternatives for Cd-based QDs in the fields of biology and medicine. However, high-quality hydrophobic CIS QDs are difficult to be transferred to water due to their 1-dodecylmercaptan (DDT) ligands. Therefore, the fluorescence and stability of the prepared aqueous CIS QDs is not enough to meet the requirement for sensitive detection. Here, as large as 13 nm CuInZnS/ZnS QDs with DDT ligands were first synthesized, and then, CuInZnS/ZnS microbeads (QBs) containing thousands of QDs were successfully fabricated by a two-step approach of emulsion-solvent evaporation and surfactant substitution. Through emulsion-solvent evaporation, the CuInZnS/ZnS QDs formed microbeads in the microemulsion with dodecyl trimethylammonium bromide (DTAB), and the Förster resonance energy transfer (FRET) has been effectively overcome. Then, CO-520 was introduced to substitute DTAB to improve the stability and water solubility. Lastly, the microbeads were coated with a SiO2 shell and carboxylated. Subsequently, the constructed QBs (∼210 nm) were used as labels in a fluorescence immunosorbent assay (FLISA) for quantitative detection of heart type fatty acid binding protein (H-FABP), and the limit of detection was 0.48 ng mL-1, which indicated a greatly improved detection sensitivity compared to that of the Cd-free QDs. The highly fluorescent and stable CuInZnS/ZnS QBs will have great application prospects in many biological fields.

An electrochemical sensor for direct and sensitive detection of ketamine

Ketamine is an organic drug with weak electrochemical activity, which makes it difficult to directly detect by electrochemical methods. Herein, an electrochemical sensor, with excellent detection sensitivity, is proposed for direct detection of ketamine based on a weakly conductive poly-L-cysteine molecularly imprinted membrane. Poly-L-cysteine molecularly imprinted membrane sensor (poly-L-Cys-KT-MIM/GCE) is obtained using L-cysteine as a functional monomer and ketamine as a template molecule based on electropolymerization. The green and highly active cysteine is selected as a functional monomer during electropolymerization, which cannot only achieve specific recognition but also improve detection sensitivity. Furthermore, the oxidation mechanism and fingerprint of ketamine on the electrode surface are established by analyzing the corresponding oxidation products using high/resolution mass spectrometry, which will help to promote the application of electrochemistry in the rapid detection of drugs. Under optimal conditions, the as-designed sensor demonstrated a linear response to ketamine within the range of 5.0 × 10−7 to 2.0 × 10−5 mol L−1 and a detection limit of 1.6 × 10−7 mol L−1. The proposed method exhibited excellent performance from the viewpoints of selectivity, sensitivity and stability. Notably, the sensor rendered excellent reliability and could be used for the detection of target analytes in hair and urine samples with high recovery rates.

Detection of copper ions in an aqueous solution by a dual-peak long period fiber grating functionalized with a polypyrrole-chitosan composite.

A dual-peak long period fiber grating (DP-LPFG) sensor functionalized by polypyrrole-chitosan composite was proposed for sensitive detection of Cu2+ ions in aqueous solution. The nitrogen atom on the polypyrrole ring and the amino group on the chitosan chain in the complex matrix can chelate the Cu2+ ions. Thus, the refractive index of the overlay changed and further modulated the transmission spectrum of DP-LPFG. After special design, the double peaks can move in opposite directions with the increase of Cu2+ ion concentration, thereby greatly improving detection sensitivity. The linear sensitivity of the fabricated sensor was measured to be 9.12 and 2.14 nm/ppm (0.61 and 0.14 nm/µM) for concentrations of 0.1-0.5 (1.5 µM-7.5 µM) and 0.5-2 ppm (7.5 µM-30 µM), respectively. In addition, the Langmuir isothermal model was used to evaluate the overall response of the sensor to Cu2+ ions quantitatively, and the detection limit was determined to be 0.05 ppb (0.75 nM). This ingenious sensor offers a new solution for sensitive detection of heavy metal ions in environmental water.

Simultaneous Sensing of Multiplex Volatile Organic Compounds by Adsorption and Plasmon Dual-Induced Raman Enhancement Technique.

Developing highly efficient gas sensors with excellent performance for rapid and sensitive detection of volatile organic compounds (VOCs) is of critical importance for the protection of human health, ecological environment, and other factors. Here, a robust gas sensor based on Raman technology was constructed by an in situ grown 2D covalent organic framework (COF) on Au nanoparticles' surface in the microchannel. Dual enhancement effects are included for the as-prepared microfluidic sensor. First, acting as a gas confinement chamber, the 2D COF could effectively capture gas molecules with high adsorption capacity and fast adsorption kinetics, resulting in VOCs' preconcentration at a high level in the COF layer. At the same time, after being stacked in the microchannel, abundant hot spots were generated among the nanogaps of Au@COF NPs. The local surface plasmon resonance effect could effectively enhance the Raman intensity. Both factors contribute to the improved detection sensitivity of VOCs. As a demonstration, several representative VOCs with different functional groups were tested. The resultant Raman spectra were subjected to the statistical principal component analysis. Varied VOCs can be successfully detected with a detection limit as low as ppb level and distinguished with 95% confidence interval. The present microfluidic platform provides a simple, sensitive, and fast method for VOCs' sensing and distinguishing, which is expected to hold potential applications in the fields of health, agricultural, and environmental research.

Correlation transmissibility damage indicator for deterioration performance analysis of hydropower generator unit

The condition monitoring system of the hydropower generator unit (HGU) contains a vast amount of monitoring data related to equipment performance. It is challenging to reveal the HGU performance change law from these data. This paper constructed a correlation transmissibility damage indicator (CTDI) to analyze HGU degradation performance based on the transmissibility function (TF). Unlike traditional TF damage indicators, CTDI can use different types of monitoring data to obtain system TF to increase TF combination and improve detection sensitivity. The analysis is performed using actual monitoring data from a hydropower plant in Guangxi, China. Firstly, a new frequency concept is defined based on the characteristics of HGU obtaining data every 10 minutes. Then, the HGU is considered a system and divided into three subsystems according to the sensor installation locations of the monitoring system. The damage indicator is used to analyze the system and its subsystems at the time scales of day, week, and month to locate and quantify the damage of HGU. The final results of this analysis show that the method can accurately detect and locate faults, applies to different operating conditions, and reveals the HGU performance deterioration pattern, which is of great significance for HGU maintenance.

A simple antigen-antibody reaction using ultrasmall FeCo nanoparticles

In this study, we prepared ultrasmall FeCo nanoparticles (NPs) with a high magnetic moment and examined their antigen-antibody reaction for biodetection applications. The FeCo NPs were collected from the FeCo-BaF2 nanogranular film with Fe:Co:Ba:F = 14:11:21:54 at.%, by dissolving the film in water since the BaF2 matrix was deliquescent. The size of FeCo NPs was ∼5 nm and the saturation magnetization was estimated to be ∼15.30 kG (149.0 emu/g). The Candida albicans antibodies (abcam ab53891)-conjugated FeCo NPs were collected by using an ultracentrifugal separation (110 000 rpm, 90 min), they were then reacted with Candida albicans. The obtained result indicates that Candida albicans were absorbed successfully onto FeCo NPs, and the number of Candida albicans bound to FeCo NPs counted from the micrographs of the aggregates of FeCo NPs and Candida albicans increased significantly by adding sonication treatment of the film in water before binding them to the antibodies. The success of antigen-antibody reaction of ultrasmall NPs with high magnetic moment improves detection sensitivity as well as offers potential detection for smaller biomolecules.

CRISPR/Cas12a-powered evanescent wave fluorescence nanobiosensing platform for nucleic acid amplification-free detection of Staphylococcus aureus with multiple signal enhancements

Although CRISPR-based biosensors for pathogenic detection are highly specific, they not sensitive enough and nucleic acid amplification is generally required to improve their sensitivity. However, this allows only binary operations and significantly limits practical applications. Here, a CRISPR/Cas12a-powered Evanescent wAve fluorescence nanobiosensing plaTform (CREAT) was developed for ultrasensitive nucleic acid amplification-free quantitative detection of pathogens with multiple signal enhancements. In addition to collateral cleavage amplification of the CRISPR/Cas12a system, we constructed nanophotonic structure-based evanescent wave fluorescence enhancement, Mg2+ or DNA-mediated fluorescence enhancement, and air-displacement fluorescence enhancement strategies for ultrasensitive detection of Staphylococcus aureus (S. aureus). Especially, the fluorescence signal detected by CREAT can be significantly enhanced by adding a simple air displacement step, thus improving detection sensitivity. This nanobiosensor detected real samples containing S. aureus, with a detection limit of 592 CFU/mL and 13.2 CFU/mL in 45 min and 90 min, respectively, which are comparable to those of RT-qPCR. This paves a new way for simple, rapid, sensitive, robust, and flexible on-site detection of S. aureus as well as other pathogens.