Label-free fluorescent biosensor based on AuNPs etching releasing signal for miRNA-155 detection.

Quantitative microRNA (miRNA) detection is crucial for early breast cancer diagnosis and prognosis. However, quick and stable fluorescence sensing for miRNA identification is still challenging. This work developed a novel label-free detection method based on AuNPs etching for quantitatively detecting miRNA-155. A layer of AuNPs was grown on the surface of mesoporous silica nanoparticles (MSN) loaded with Rhodamine 6G (R6G) using seed-mediated growth, followed by probe attachment. In the presence of miRNA-155, the MSN@R6G@AuNP surface loses the protection of the attached probe, rendering AuNPs susceptible to etching by hydrochloric acid. This results in a significant fluorescent signal being released in the free space. The encapsulation with AuNPs effectively reduces signal leakage, while the rapid etching process shortens detection time. This strategy enables sensitive and fast detection with a detection range of 100 fM to 100 nM, a detection limit of 2.18 fM, and a detection time of 30 min. The recovery rate in normal human serum ranges from 99.02 % to 106.34 %. This work presents a simple biosensing strategy with significant potential for application in tumor diagnosis.

A universal electrochemical biosensor based on CRISPR/Cas12a and a DNA tetrahedron for ultrasensitive nucleic acid detection.

Herein, a universal nucleic acid analysis platform was constructed for sensitive and accurate detection of miRNA-155 and ctDNA using isothermal amplification-assisted CRISPR/Cas12a and a tetrahedral DNA nanostructure (TDN) supported sensing interface. Under the optimal experimental conditions, the prepared sensor achieved specific detection of miRNA-155 and ctDNA at as low as aM levels in 2.6 h. Furthermore, the platform was also successfully applied to human serum sample recovery experiments and cancer cell lysates, demonstrating outstanding reliability and accuracy. We firmly believe that this work provides a universal, sensitive, and practical tool for early clinical diagnosis.

A sensitive sandwich-type electrochemical aptasensing platform based on Ti3C2Tx/MoS2/MWCNT@rGONR composites for simultaneous detection of kanamycin and chloramphenicol in food samples.

A Ti3C2Tx/MoS2/MWCNT@rGONR nanocomposite was prepared for the first time for building a sensitive electrochemical aptasening platform to simultaneously detect kanamycin (Kana) and chloramphenicol (Cap). Owing to their accordion-like structure, rich surface groups, and high charge mobility, Ti3C2Tx/MoS2/MWCNT@rGONR composites provided a spacious covalent immobilization surface and a better electrochemical aptasensing platform. The aptamers of Kana and Cap used in sensors enhance the selectivity. Furthermore, TiP, an ion exchanger, was used for loading more different metal ions functioning as labels to form a sandwich-type sensor together with Ti3C2Tx/MoS2/MWCNT@rGONR, improving the electrochemical sensitivity and obtaining a highly distinguishable signal readout. Under the optimized conditions, the sensor has good detection limits of 0.135 nmol L-1 and 0.173 nmol L-1 for Kana and Cap, respectively, at the same linearity concentration of 0.5-2500 nmol L-1. Finally, it was successfully applied for detection in milk and fish meat, and the results were compared with the standard method HPLC, indicating its great potential for food safety monitoring.

Histidine modified Fe3O4 nanoparticles improving the ethanol yield and tolerance of Saccharomyces cerevisiae.

Saccharomyces cerevisiae, the primary microorganism involved in ethanol production, is hindered by the accumulation of ethanol, leading to reduced ethanol production. In this study, we employed histidine-modified Fe3O4 nanoparticles (His-Fe3O4) for the first time, to the best of our knowledge, as a method to enhance ethanol yield during the S. cerevisiae fermentation process. The results demonstrated that exposing S. cerevisiae cells to Fe3O4 nanoparticles (Fe3O4 NPs) led to increased cell proliferation and glucose consumption. Moreover, the introduction of His-Fe3O4 significantly boosted ethanol content by 17.3% (p < 0.05) during fermentation. Subsequent findings indicated that the increase in ethanol content was associated with enhanced ethanol tolerance and improved electron transport efficiency. This study provided evidence for the positive effects of His-Fe3O4 on S. cerevisiae cells and proposed a straightforward approach to enhance ethanol production in S. cerevisiae fermentation. The mediation of improved ethanol tolerance offers significant potential in the fermentation and bioenergy sectors.

Rapid, portable, and sensitive detection of CaMV35S by RPA-CRISPR/Cas12a-G4 colorimetric assays with high accuracy deep learning object recognition and classification.

Fast, sensitive, and portable detection of genetic modification contributes to agricultural security and food safety. Here, we developed RPA-CRISPR/Cas12a-G-quadruplex colorimetric assays that can combine with intelligent recognition by deep learning algorithms to achieve sensitive, rapid, and portable detection of the CaMV35S promoter. When the crRNA-Cas12a complex recognizes the RPA amplification product, Cas12 cleaves the G-quadruplex, causing the G4-Hemin complex to lose its peroxide mimetic enzyme function and be unable to catalyze the conversion of ABTS2- to ABTS, allowing CaMV35S concentration to be determined based on ABTS absorbance. By utilizing the RPA-CRISPR/Cas12a-G4 assay, we achieved a CaMV35S limit of detection down to 10 aM and a 0.01 % genetic modification sample in 45 min. Deep learning algorithms are designed for highly accurate classification of color results. Yolov5 objective finding and Resnet classification algorithms have been trained to identify trace (0.01 %) CaMV35S more accurately than naked eye colorimetry. We also coupled deep learning algorithms with a smartphone app to achieve portable and rapid photo identification. Overall, our findings enable low cost ($0.43), high accuracy, and intelligent detection of the CaMV35S promoter.

Label-Free and DNAzyme-Mediated Biosensor with a High Signal-to-Noise Ratio for a Lumpy Skin Disease Virus Assay.

Lumpy skin disease virus (LSDV) is a severe and highly contagious form of cowpox. As LSDV continues to mutate and there is no vaccine and treatment in nonendemic countries, early detection of LSDV becomes an important basis for epidemic prevention and control, especially for detection of conserved sequences. A new label-free and sensitive fluorescence method was developed based on a light-up RNA aptamer for detecting LSDV. The method integrated recombinase polymerase amplification (RPA), CRISPR/Cas12a, 10-23 DNAzyme, and Baby Spinach RNA aptamer for triple cascade signal amplification. Based on highly sensitive and specific RPA and CRISPR/Cas12a, DNAzyme achieved a third signal amplification. Additionally, the Baby Spinach RNA aptamer had stronger fluorescence signals and higher quantum yields. The label-free method had ultrahigh sensitivity with the actual detection limit as 1.29 copies·µL-1. The method was 100-fold more sensitive compared to RPA with Cas12a. Moreover, it had no cross-reactivity with viruses belonging to the Capripoxvirus, such as sheep pox virus and goat pox virus with genetic homology as 97%. Furthermore, the method displayed 100% accuracy in 50 actual samples. Therefore, the method based on RPA, Cas12a, and 10-23 DNAzyme had advantages in LSDV detection and provided a new solution for LSD prevention and control.

Simultaneous Detection of Micro-RNAs by a Disposable Biosensor via the Click Chemistry Connection Strategy.

Simultaneous detection of multiple breast cancer-associated miRNAs significantly raises the accuracy and reliability of early diagnosis. In this work, disposable carbon fiber paper serves as the biosensing interface, linking DNA probes via click chemistry to efficiently capture targets and signals efficiently. DNA probes have multiple recognition domains that trigger a cascade reaction through the helper probes and targets, resulting in two signals output. The signals are centrally encapsulated in the pore of the MIL-88(Fe)-NH2. The signal carriers are directed by signal probes to the recognition domains that correspond to the DNA probes. The biosensor is selective and stable, and it can quantify miRNA-21 and miRNA-155 simultaneously with detection limits of 0.64 and 0.54 fmol/L, respectively. Furthermore, it demonstrates satisfactory performance in tests conducted with normal human serum and cell lysate. Overall, this method makes a satisfactory exploration to realize an inexpensive and sensitive biosensor for multiple biomarkers.

Synergistic enhancement of wearable biosensor through Pt single-atom catalyst for sweat analysis.

Real-time monitoring of biological markers in sweat is a valuable tool for health assessment. In this study, we have developed an innovative wearable biosensor for precise analysis of glucose in sweat during physical activities. The sensor is based on a single-atom catalyst of platinum (Pt) uniformly dispersed on tricobalt tetroxide (Co3O4) nanorods and reduced graphene oxide (rGO), featuring a unique three-dimensional nanostructure and excellent glucose electrocatalytic performance with a wide detection range of 1-800 µM. Additionally, density functional theory calculations have revealed the synergetic role of Pt active sites in the Pt single-atom catalyst (Co3O4/rGO/Pt) in glucose adsorption and electron transfer, thereby enhancing sensor performance. To enable application in wearable devices, we designed an S-shaped microfluidic chip and a point-of-care testing (POCT) device, both of which were validated for effectiveness through actual use by volunteers. This research provides valuable insights and innovative approaches for analyzing sweat glucose using wearable devices, contributing to the advancement of personalized healthcare.

A flexible self-supported electrochemical sensor Co-NC/PS@CC for real-time detection of cell-released H2O2.

BACKGROUND: Hydrogen peroxide (H2O2) is an important reactive oxygen species (ROS) molecule involved in cell metabolism regulation, transcriptional regulation, and cytoskeleton remodeling. Real-time monitoring of H2O2 levels in live cells is of great significance for disease prevention and diagnosis. RESULTS: We utilized carbon cloth (CC) as the substrate material and employed a single-atom catalysis strategy to prepare a flexible self-supported sensing platform for the real-time detection of H2O2 secreted by live cells. By adjusting the coordination structure of single-atom sites through P and S doping, a cobalt single-atom nanoenzyme Co-NC/PS with excellent peroxidase-like activity was obtained. Furthermore, we explored the enzyme kinetics and possible catalytic mechanism of Co-NC/PS. Due to the excellent flexibility, high conductivity, strong adsorption performance of carbon cloth, and the introduction of non-metallic atom-doped active sites, the developed Co-NC/PS@CC exhibited ideal sensing performance. Experimental results showed that the linear response range for H2O2 was 1-17328 µM, with a detection limit (LOD) of 0.1687 µM. Additionally, the sensor demonstrated good reproducibility, repeatability, anti-interference, and stability. SIGNIFICANCE: The Co-NC/PS@CC prepared in this study has been successfully applied for detecting H2O2 secreted by MCF-7 live cells, expanding the application of single-atom nanoenzymes in live cell biosensing, with significant implications for health monitoring and clinical diagnostics.

A Novel CRISPR/Cas12a-Mediated Ratiometric Dual-Signal Electrochemical Biosensor for Ultrasensitive and Reliable Detection of Circulating Tumor Deoxyribonucleic Acid.

Circulating tumor DNA (ctDNA) holds great promise as a noninvasive biomarker for cancer diagnosis, treatment, and prognosis. However, the accurate and specific quantification of low-abundance ctDNA in serum remains a significant challenge. This study introduced, for the first time, a novel exponential amplification reaction (EXPAR)-assisted CRISPR/Cas12a-mediated ratiometric dual-signal electrochemical biosensor for ultrasensitive and reliable detection of ctDNA. To implement the dual-signal strategy, a signal unit (ssDNA-MB@Fc/UiO-66-NH2) was prepared, consisting of methylene blue-modified ssDNA as the biogate to encapsulate ferrocene signal molecules within UiO-66-NH2 nanocarriers. The presence of target ctDNA KRAS triggered EXPAR amplification, generating numerous activators for Cas12a activation, resulting in the cleavage of ssDNA-P fully complementary to the ssDNA-MB biogate. Due to the inability to form a rigid structure dsDNA (ssDNA-MB/ssDNA-P), the separation of ssDNA-MB biogate from the UiO-66-NH2 surface was hindered by electrostatic interactions. Consequently, the supernatant collected after centrifugation exhibited either no or only a weak presence of Fc and MB signal molecules. Conversely, in the absence of the target ctDNA, the ssDNA-MB biogate was open, leading to the leakage of Fc signal molecules. This clever ratiometric strategy with Cas12a as the "connector", reflecting the concentration of ctDNA KRAS based on the ratio of the current intensities of the two electroactive signal molecules, enhanced detection sensitivity by at least 60-300 times compared to single-signal strategies. Moreover, this strategy demonstrated satisfactory performance in ctDNA detection in complex human serum, highlighting its potential for cancer diagnosis.

Label-free fluorescent sensor for sensitive detection of ctDNA based on water stabilized CsPbBr3 nanosheet.

The detection of circulating tumor DNA (ctDNA), as a practical liquid biopsy technique, was of great significance for the study of cancer diagnosis and prognosis. However, reported methods for detection ctDNA still have some limitations, such as tedious process and high cost. In this study, CsPbBr3 nanosheet (CsPbBr3 NS) with high water stability was prepared by etching, and its fluorescence intensity could be stably stored for 1 year. The Ti3C2Tx possessed high quenching efficiency for CsPbBr3 NS and the HOMO-LUMO orbital study revealed that the PET mechanism was responsible for fluorescence quenching. And the Ti3C2Tx showed stronger affinity towards single-stranded DNA (ssDNA), as compared with double-stranded DNA (dsDNA). The probe ssDNA could be adsorbed on the surface of Ti3C2Tx through π-π stacking. After the targets were recognized by probe ssDNA to form dsDNA, its affinity with Ti3C2Tx decreased and the active site of Ti3C2Tx recovered, causing a high quenching efficiency on CsPbBr3 NS. Based on this, a label-free fluorescent biosensor was designed for the sensitive detection of ctDNA (EGFR 19 Dels for non-small cell lung cancer, NSCLC). Under the optimal experimental conditions, this biosensor exhibited a detection limit of 180 fM and a linear range of 50 pM-350 pM with amplification of magnetic beads through strand displacement reaction. In addition, this sensor was applied to the detection of ctDNA in serum samples and cells lysates. This method for ctDNA detection was expected to have great potential for biomarker detection in the field of liquid biopsy.

Fe Single-Atom Carbon Dots Nanozyme Collaborated with Nucleic Acid Exonuclease III-Driven DNA Walker Cascade Amplification Strategy for Circulating Tumor DNA Detection.

Circulating tumor DNA (ctDNA), as a next-generation tumor marker, enables early screening and monitoring of cancer through noninvasive testing. Exploring the development of new methods for ctDNA detection is an intriguing study. In this work, a unique electrochemical biosensor for the ctDNA detector was constructed in the first utilizing Fe single-atom nanozymes-carbon dots (SA Fe-CDs) as a signaling carrier in collaboration with a DNA walker cascade amplification strategy triggered by nucleic acid exonuclease III (Exo III). The electrochemical active surface area of AuNPs/rGO modified onto a glassy carbon electrode (AuNPs/rGO/GCE) was about 1.43 times that of a bare electrode (bare GCE), with good electrical conductivity alongside a high heterogeneous electron transfer rate (5.81 × 10-3 cm s-1), that is, as well as the ability to load more molecules. Sequentially, the DNA walker cascade amplification strategy driven by Exo III effectively converted the target ctDNA into an amplified biosignal, ensuring the sensitivity and specificity of ctDNA. Ultimately, the electrochemical signal was further amplified by introducing SA Fe-CDs nanozymes, which could serve as catalysts for 3,3',5,5'-tetramethylbenzidine (TMB) oxidation with facile responding (Vmax = 0.854 × 10-6 M s-1) and robust annexation (Km = 0.0069 mM). The integration of the triple signal amplification approach achieved detection limits as low as 1.26 aM (S/N = 3) for a linearity spanning from 5 aM to 50 nM. In this regard, our proposal for a biosensor with exceptional assay properties in complicated serum environments had great potential for early and timely diagnosis of cancer.

A wearable flexible electrochemical biosensor with CuNi-MOF@rGO modification for simultaneous detection of uric acid and dopamine in sweat.

BACKGROUND: In health assessment and personalized medical services, accurate detection of biological markers such as dopamine (DA) and uric acid (UA) in sweat is crucial for providing valuable physiological information. However, there are challenges in detecting sweat biomarkers due to their low concentrations, variations in sweat yield among individuals, and the need for efficient sweat collection. RESULTS: We synthesized CuNi-MOF@rGO as a high-activity electrocatalyst and investigated its feasibility and electrochemical mechanism for simultaneously detecting low-concentration biomarkers UA and DA. Interaction between the non-coordinating carboxylate group and the sample produces effective separation signals for DA and UA. The wearable biomimetic biosensor has a wide linear range of 1-500 µM, with a detection limit of 9.41 µM and sensitivity of 0.019 µA µM-1 cm-2 for DA, and 10-1000 µM, with a detection limit of 9.09 µM and sensitivity of 0.026 µA µM-1 cm-2 for UA. Thus, our sensor performs excellently in detecting low-concentration biomarkers. To improve sweat collection, we designed a microfluidic-controlled device with hydrophilic modification in the microchannel. Experimental results show optimal ink flow at 2% concentration. Overall, we developed an innovative and highly active electrocatalyst, successfully enabling simultaneous detection of low-concentration biomarkers UA and DA. SIGNIFICANCE: This study provides a strategy for sweat analysis and health monitoring. Moreover, the sensor also showed good performance in detecting real sweat samples. This study has shown great potential in future advances in sweat analysis and health monitoring.

Sulfhydryl-functionalized 3D MXene-AuNPs enabled electrochemical sensors for the selective determination of Pb2+, Cu2+ and Hg2+ in grain.

Herein, we made 3D MXene-AuNPs by in situ growth of gold nanoparticles (AuNPs) on the surface of MXene by chemical reduction method, and then introduced three sulfhydryl (-SH) compounds as functionalized modifiers attached to the AuNPs to form a highly selective composite material for the detection of Pb2+, Cu2+, and Hg2+, respectively. The doping of AuNPs changes the microstructure of 2D MXene and generates more active sites. On a sensing platform based on ITO array electrodes, the detection system was optimised with sensitivities up to 1.157, 0.846 and 0.799 µA·µg-1Lcm-2 (Pb2+, Cu2+, and Hg2+). The selectivity of MXene@AuNPs was effectively improved by sulfhydryl group modification. In the range of 1-1300 µg L-1, the detection limits of three ions were 0.07, 0.13 and 0.21 µg L-1. In addition, this method can efficiently and accurately detect heavy metal ions in four cereal samples with consistent results with inductively coupled plasma mass spectrometry.

Pattern-based colorimetric sensor array chip for discrimination of Baijiu aromas.

Gas mixtures are comprised of numerous complex components, making the accurate identification a continuing challenge due to the significant limitations of existing detection methods. Herein, we developed a low-cost and sensitive pattern-based colorimetric sensor array chip for the identification of typical gas mixtures - Baijiu aroma. Specifically, three nanomaterials (AuNPs, MoS2 and ZIF-8) were prepared to adsorb gas molecules and enhance the reaction of trace gases with sensor arrays. The colorimetric sensor array chip took only 5 min to complete the recognition of Baijiu aromas and effectively avoided recognition errors caused by sommelier olfactory fatigue. Notably, the hierarchical cluster analysis (HCA) revealed no confusion or errors in the results of 80 tests across the five trials involving 16 commercial Baijius. Even fake Baijius with similar ingredients could be easily identified, demonstrating the excellent analytical capabilities of the system in Baijiu identification and its significant potential for quality control of Baijius.

Wearable electrochemical patch based on iron nano-catalysts incorporated laser-induced graphene for sweat metabolites detection.

The development of wearable devices shows great application potential in health management. In this work, we propose the fabrication of a novel wearable electrochemical patch and prove its application in sweat metabolites detection. The patch is developed based on iron nano-catalysts incorporated laser-induced graphene (FeNCs/LIG), which is a newly integrated sensing electrode with unique three-dimensional nanostructure and good electrocatalytic activity. It shows desirable sensing performances for sweat metabolites including tyrosine (Tyr) and uric acid (UA) molecules. The detection limit of Tyr and UA can reach 5.11 µM and 1.37 µM, respectively. Besides, density functional theory calculation deeply reveals that the Fe active sites of FeNCs play an important role in molecule adsorption and electron transference, thus promoting sensing performance. To realize wearable application, a dual-channel hydrogel chip is designed and assembled with FeNCs/LIG. The developed patch is successfully utilized to accurately determination of Tyr and UA in sweat. This work is expected to provide a new non-invasive strategy for evaluating amino acid intake and metabolic level.

Ultra-sensitive electrochemical sensors through self-assembled MOF composites for the simultaneous detection of multiple heavy metal ions in food samples.

Using an assemble-able MOF material, we successfully constructed an ultra-sensitive electrochemical sensor based on Bi2CuO4@Al-MOF@UiO-67 nanocomposite material, in order to investigate the adsorption properties of the Bi2CuO4@Al-MOF@UiO-67 functional material on the heavy metal ion. The Cd2+, Cu2+, Pb2+ and Hg2+ can be detected at the same time. Selective recognition and enrichment of various metal ions on different substrates can be achieved through the assembly of a large number of Al-MOF and UiO-67-MOF nanomaterial composites with small particle sizes on the Bi2CuO4 surface. Based on this, a new type of sensor is researched and prepared, which has been shown to have good stability and reproducibility. Due to its unique assembly structure, large active surface area, excellent adsorption capacity, and high electrical conductivity, Bi2CuO4@Al-MOF@UiO-67 presents outstanding performance. In addition, the sensor also exhibits excellent electrocatalytic redox capacity and high selectivity. The adsorption capacity of Cd2+, Cu2+, Pb2+ and Hg2+ is also significantly improved under the action of the sensor electrode, however, this is not the case. The limits of detection for Cd2+, Cu2+, Pb2+ and Hg2+ were found to be 0.02 pM, 0.032 pM, 0.018 pM and 0.041 pM, respectively. In order to investigate the detection mechanism of Cd2+, Cu2+, Pb2+ and Hg2+ was adsorption properties as well as electrochemical accumulation of Bi2CuO4@Al-MOF@UiO-67 on the metal atoms were investigated. This method has been successfully applied to samples of rice, sorghum, maize, milk, honey, and tea, and has enabled the simultaneous detection of Cd2+, Cu2+, Pb2+ and Hg2+, which is of significant practical value.

A three-modal fluorescent sensor harnessing diverse luminescent mechanisms for the purpose of segmented Baijiu identification.

The classification and verification of segmented Baijiu hold significant importance as they profoundly influence the blending and overall quality of the Baijiu. Our scholarly investigation yielded a fluorescent sensor with three luminescent modes by integrating Tb3+ and RHB into UiO-66. The interplay between carboxyl-containing compounds and RHB/Tb@TLU-2 orchestrates a harmonious molecular association, where the convergence of carboxyl groups with Tb3+ yields a resonating impact on the antenna effect of BDC-SO3-. Furthermore, the acidity and alkalinity of reactants induced a charge transfer interaction between BDC-NH2 and Zr4+ and led to structural changes in RHB/Tb@TLU-2, resulting in observable fluorescence signal variations across the three emission centers. The sensor array successfully identified eight organic acids, achieving an impressive 97.5 % accuracy in discerning segmented Baijiu samples from four Baijiu pits. This meticulous methodology prioritizes simplicity, swiftness, and effectiveness, paving the path for comprehensive segmented Baijiu analysis in the esteemed realm of Brewing production.

A novel colorimetric and fluorometric dual-signal identification of organics and Baijiu based on nanozymes with peroxidase-like activity.

Nanozymes were nanomaterials with enzymatic properties. They had diverse functions, adjustable catalytic activity, high stability, and easy large-scale production, attracting interest in biosensing. However, nanozymes were scarcely applied in Baijiu identification. Herein, a colorimetric and fluorometric dual-signal determination mediated by a nanozyme-H2O2-TMB system was developed for the first time to identify organics and Baijiu. Since the diverse peroxidase-like activity of nanozymes, resulted in different degrees of oxidized TMB. Based on this, 21 organics were identified qualitatively and quantitatively by colorimetric method with a rapid response (<12 min), broad linearity (0.0005-35 mM), and low detection limits (a minimum of 30 nM for glutaric acids). Furthermore, the fluorometric method exhibited excellent potential for accurate determination of organics, with detection ranges of 2-200 µmol/L (LOD: 0.22 µmol/L) for l-ascorbic acid and 2-300 µmol/L (LOD: 0.59 µmol/L) for guaiacol. Finally, the sensor was successfully applied to identify fake Baijiu and Baijiu from 16 different brands.