CLICK-FLISA Based on Metal-Organic Frameworks for Simultaneous Detection of Fumonisin B1 (FB1) and Zearalenone (ZEN) in Maize.

Mycotoxins are secondary products produced primarily by fungi and are pathogens of animals and cereals, not only affecting agriculture and the food industry but also causing great economic losses. The development of rapid and sensitive methods for the detection of mycotoxins in food is of great significance for livelihood issues. This study employed an amino-functionalized zirconium luminescent metal-organic framework (LOF) (i.e., UiO-66-NH2). Click chemistry was utilized to assemble UiO-66-NH2 in a controlled manner, generating LOF assemblies to serve as probes for fluorescence-linked immunoassays. The proposed fluoroimmunoassay method for Zearalenone (ZEN) and Fumonisin B1 (FB1) detection based on the UiO-66-NH2 assembled probe (CLICK-FLISA) afforded a linear response range of 1-20 µmol/L for ZEN, 20 µmol/L for FB1, and a very low detection limit (0.048-0.065 µmol/L for ZEN; 0.048-0.065 µmol/L for FB1). These satisfying results demonstrate promising applications for on-site quick testing in practical sample analysis. Moreover, the amino functionalization may also serve as a modification strategy to design luminescent sensors for other food contaminants.

Micro-/nanostructures for surface-enhanced Raman spectroscopy: Recent advances and perspectives.

Surface-enhanced Raman spectroscopy (SERS) has great potential for the analysis of molecules adsorbed on metals with rough surfaces or substrates with micro-/nanostructures. Plasmonic coupling between metal nanoparticles and the morphology of the rough metal surface can produce "hot spots" that enhance Raman scattering by adsorbed molecules, typically at micro- to nanomolar concentrations, although high enhancement factors can also facilitate single-molecule detection. This phenomenon is widely applicable for chemical analysis and sensing in various fields. In this review, the latest research progress on SERS micro-/nanosensors is evaluated, and the sensors are classified according to their individual functions. Furthermore, the design principles and working mechanisms of reported SERS-active micro-/nanostructured substrates are analyzed, and the design features adopted to overcome the difficulties associated with precision detection are explored. Finally, challenges and directions for future development in this field are discussed. This review serves as a design guide for novel SERS-active substrates.

Synthetic biology for the food industry: advances and challenges.

As global environmental pollution increases, climate change worsens, and population growth continues, the challenges of securing a safe, nutritious, and sustainable food supply have become enormous. This has led to new requirements for future food supply methods and functions. The use of synthetic biology technology to create cell factories suitable for food industry production and renewable raw material conversion into: important food components, functional food additives, and nutritional chemicals, represents an important method of solving the problems faced by the food industry. Here, we review the recent progress and applications of synthetic biology in the food industry, including alternatives to: traditional (artificial pigments, meat, starch, and milk), functional (sweeteners, sugar substitutes, nutrients, flavoring agents), and green (green fiber, degradable packing materials, green packaging materials and food traceability) foods. Furthermore, we discuss the future prospects of synthetic biology-based applications in the food industry. Thus, this review may serve as a reference for research on synthetic biology in the: food safety, food nutrition, public health, and health-related fields.

A facile dual-mode immunosensor based on speckle Ag-doped nanohybrids for ultrasensitive detection of Ochratoxin A.

Ochratoxin A (OTA) is a potent carcinogen, and is among the most dangerous mycotoxins in agricultural products. In this study, an ultrasensitive dual-mode immunosensor was developed for naked-eye and fluorescence detection of OTA based on Ag-doped core-shell nanohybrids (Ag@CSNH). Complete antigen-labeled Ag@CSNH (CA-Ag@CSNH) were used as a competitive bind and dual-mode probe. The diffused doping structure of CA-Ag@CSNH provided improved stability, color and fluorescence quencher performance. Antibodies modified magnetic beads were used as a capture probe. The competitive binding between OTA and CA-Ag@CSNH produced both color change and fluorescence quenching. Ultraviolet and fluorescence intensitie correlated linearly with OTA concentration ranges of 0.03-3 ng/mL and 10-10000 pg/mL, and limits of detection of 0.0235 ng/mL and 0.9921 pg/mL, respectively. The practical applicability of proposed strategy was demonstrated by analysis of OTA in spiked corn, soybean and flour samples. This study offers a new insight on multi-mode platforms for various applications.

Functionalized magnetic nanobeads for SERS-based detection of Staphylococcus aureus.

In this study, we introduced a Raman detection technique based on a combination of functionalized magnetic beads and surface-enhanced Raman scattering (SERS) tags to develop a rapid and sensitive strategy for the detection of Staphylococcus aureus (S. aureus), a typical foodborne pathogen. Polyethylene glycol (PEG) and bovine serum albumin (BSA) dual-mediated teicoplanin functionalized magnetic beads (TEI-BPBs) were prepared for separation of target bacteria. SERS tags were used to immobilize antibodies on gold surfaces with bifunctional linker proteins to ensure specific recognition of S. aureus. Under optimal conditions, the combination of TEI-BPBs and SERS tags showed reliable performance, exhibiting good capture efficiency even in the presence of 106 CFU mL-1 of non-target bacteria. The SERS tag provided an effective hot spot for subsequent Raman detection, presenting good linearity in the range of 102-107 CFU mL-1. Good performance has also been shown in detecting target bacteria in milk samples, where it has a recovery of 95.5-101.3%. Thus, the highly sensitive Raman detection technique combined with TEI-BPBs capture probes and SERS tags is a promising method for the detection of foodborne pathogens in food or clinical samples.

The development of a fluorescence/colorimetric biosensor based on the cleavage activity of CRISPR-Cas12a for the detection of non-nucleic acid targets.

Nano-biosensors are of great significance for the analysis and detection of important biological targets. Surprisingly, the CRISPR-Cas12a system not only provides us with excellent gene editing capabilities, it also plays an important role in biosensing due to its high base resolution and high levels of sensitivity. However, most CRISPR-Cas12a-based sensors are limited by their recognition and output modes, are therefore only utilized for the detection of nucleic acids using fluorescence as an output signal. In the present study, we further explored the potential application of CRISPR-Cas12a and developed a CRISPR-Cas12a-based fluorescence/colorimetric biosensor (UCNPs-Cas12a/hydrogel-MOF-Cas12a) that provides an efficient targeting system for small molecules and protein targets. These two sensors yield multiple types of signal outputs by converting the target molecule into a deoxyribonucleic acid (DNA) signal input system using aptamers, amplifying the DNA signal by catalyzed hairpin assembly (CHA), and then combining CRISPR-Cas12a with various nanomaterials. UCNPs-Cas12a/hydrogel-MOF-Cas12a exhibited prominent sensitivity and stability for the detection of estradiol (E2) and prostate-specific antigen (PSA), and was successfully applied for the detection of these targets in milk and serum samples. A major advantage of the hydrogel-MOF-Cas12a system is that the signal output can be observed directly. When combined with aptamers and nanomaterials, CRISPR-Cas12a can be used to target multiple targets, with a diverse array of signal outputs. Our findings create a foundation for the development of CRISPR-Cas12a-based technologies for application in the fields of food safety, environmental monitoring, and clinical diagnosis.

AuNP/Cu-TCPP(Fe) metal-organic framework nanofilm: a paper-based electrochemical sensor for non-invasive detection of lactate in sweat.

Effective and real-time detection of lactate (LA) content in human sweat has attracted considerable attention from researchers. In this work, a novel electrochemical paper-based analysis device (ePAD) was developed for the non-invasive detection of LA in sweat. The electrocatalytic properties of AuNP/Cu-TCPP(Fe) hybrid nanosheets, which were prepared by an optimised synthetic method, were studied by CV and EIS electrochemical methods for the first time and the working electrode can be fabricated using a drip coating method. The lactate sensor was optimised and validated for usability, adoptability and interpretability. To the best of our knowledge, this was the fastest, lowest detection line and widest linear range method reported to date for the detection of lactate. It achieved the detection limit of 0.91 pM and a linear range from 0.013 nM to 100 mM. The dual catalytic effects of the hybrid NSs shortened the detection time by nearly two times and enhanced the sensitivity approximately two times, an accuracy unmatched until now. Furthermore, this sensor was employed for LA analysis and validated by high performance liquid chromatography (HPLC). The ePAD shows superior biocompatibility, accuracy, and high sensitivity and can be easily manufactured. Hence, it is applicable for the long-term monitoring of sweat LA concentrations in point-of-care testing, athletic testing of athletes and military personnel and other subjects in different extreme environments.

Amplification-free detection of HBV DNA mediated by CRISPR-Cas12a using surface-enhanced Raman spectroscopy.

Nucleic acid markers have been widely used in the detection of various virus-related diseases, including hepatitis B virus (HBV), which is spreading worldwide. The trans-activated CRISPR-Cas system has shown excellent sensitivity and specificity in nucleic acid detection. However, nucleic acid testing usually requires amplification of the target nucleic acid for more accurate and specific detection; furthermore, current nucleic acid assays are time-consuming, costly, and are limited by non-specific cross-reactivity. We developed an amplification-free viral DNA biosensor-based diagnostic method that uses a clustered regularly interspaced short palindromic repeats-associated system (CRISPR/Cas)-based approach with surface enhanced Raman spectroscopy. This method can specifically identify the target site by changing the crRNA sequence. In addition, the incubation period and development of the disease can be determined by quantitative detection of viral DNA. This system could achieve rapid and highly sensitive detection of HBV DNA within 50 min and vast detection range from 0.1 pM to 1 nM. Therefore, a combined CRISPR/Cas12a-SERS-based assay would improve the sensitivity of detection in assays using multiple biomarkers. In conclusion, our CRISPR/Cas12a-based biosensor would enable rapid, simple, and sensitive detection of HBV nucleic acids.

Wearable biosensors for human fatigue diagnosis: A review.

Fatigue causes deleterious effects to physical and mental health of human being and may cause loss of lives. Therefore, the adverse effects of fatigue on individuals and the society are massive. With the ever-increasing frequency of overtraining among modern military and sports personnel, timely, portable and accurate fatigue diagnosis is essential to avoid fatigue-induced accidents. However, traditional detection methods require complex sample preparation and blood sampling processes, which cannot meet the timeliness and portability of fatigue diagnosis. With the development of flexible materials and biosensing technology, wearable biosensors have attracted increased attention to the researchers. Wearable biosensors collect biomarkers from noninvasive biofluids, such as sweat, saliva, and tears, followed by biosensing with the help of biosensing modules continuously and quantitatively. The detection signal can then be transmitted through wireless communication modules that constitute a method for real-time understanding of abnormality. Recent developments of wearable biosensors are focused on miniaturized wearable electrochemistry and optical biosensors for metabolites detection, of which, few have exhibited satisfactory results in medical diagnosis. However, detection performance limits the wide-range applicability of wearable fatigue diagnosis. In this article, the application of wearable biosensors in fatigue diagnosis has been discussed. In fact, exploration of the composition of different biofluids and their potential toward fatigue diagnosis have been discussed here for the very first time. Moreover, discussions regarding the current bottlenecks in wearable fatigue biosensors and the latest advancements in biochemical reaction and data communication modules have been incorporated herein. Finally, the main challenges and opportunities were discussed for wearable fatigue diagnosis in the future.

Biodegradable Microneedle Array-Mediated Transdermal Delivery of Dimethyloxalylglycine-Functionalized Zeolitic Imidazolate Framework-8 Nanoparticles for Bacteria-Infected Wound Treatment.

Bacteria-infected skin wounds caused by external injuries remain a serious challenge to the whole society. Wound healing dressings, with excellent antibacterial activities and potent regeneration capability, are increasingly needed clinically. Here, we reported a novel functional microneedle (MN) array comprising methacrylated hyaluronic acid (MeHA) embedded with pH-responsive functionalized zeolitic imidazolate framework-8 (ZIF-8) nanoparticles to treat bacteria-infected cutaneous wounds. Antibacterial activity was introduced into Zn-ZIF-8 to achieve sterilization through releasing Zn ions, as well as increased angiogenesis by dimethyloxalylglycine (DMOG) molecules that were distributed within its framework. Furthermore, biodegradable MeHA was chosen as a substrate material carrier to fabricate DMOG@ZIF-8 MN arrays. By such design, DMOG@ZIF-8 MN arrays would not only exhibit excellent antibacterial activity against pathogenic bacteria but also enhance angiogenesis within wound bed by upregulating the expression of HIF-1α, leading to a significant therapeutic efficiency on bacteria-infected cutaneous wound healing. Based on these results, we conclude that this new treatment strategy can provide a promising alternative for accelerating infected wound healing via effective antibacterial activity and ameliorative angiogenesis.

Sensitive colorimetric detection of antibiotic resistant Staphylococcus aureus on dairy farms using LAMP with pH-responsive polydiacetylene.

Rapidly and accurately detecting antibiotic-resistant pathogens in agriculture and husbandry is important since these represent a major threat to public health. While much attention has been dedicated to detecting now-common resistant bacteria, such as methicillin-resistant Staphylococcus aureus, fewer methods have been developed to assess resistance against macrolides in Staphylococcus aureus (SA). Here, we report a visual on-site detection system for macrolide resistant SA in dairy products. First, metagenomic sequencing in raw milk, cow manure, water and aerosol deposit collected from dairy farms around Tianjin was used to identify the most abundant macrolide resistance gene, which was found to be the macB gene. In parallel, SA housekeeping genes were screened to allow selective identification of SA, which resulted in the selection of the SAOUHSC_01275 gene. Next, LAMP assays targeting the above-mentioned genes were developed and interpreted by agarose gel electrophoresis. For on-site application, different pH-sensitive colorimetric LAMP indicators were compared, which resulted in selection of polydiacetylene (PDA) as the most sensitive candidate. Additionally, a semi-quantitative detection could be realized by analyzing the RGB information via smartphone with a LOD of 1.344 × 10-7 ng/µL of genomic DNA from a milk sample. Finally, the proposed method was successfully carried out at a real farm within 1 h from sample to result by using freeze-dried reagents and portable devices. This is the first instance in which PDA is used to detect LAMP products, and this generic read-out system can be expanded to other antibiotic resistant genes and bacteria.

Ultrasensitive Automatic Detection of Small Molecules by Membrane Imaging of Single Molecule Assays.

Determination of trace amounts of targets or even a single molecule target has always been a challenge in the detection field. Digital measurement methods established for single molecule counting of proteins, such as single molecule arrays (Simoa) or dropcast single molecule assays (dSimoa), are not suitable for detecting small molecule, because of the limited category of small molecule antibodies and the weak signal that can be captured. To address this issue, we have developed a strategy for single molecule detection of small molecules, called small molecule detection with single molecule assays (smSimoa). In this strategy, an aptamer is used as a recognition element, and an addressable DNA Nanoflower (DNF) attached on the magnetic beads surface, which exhibit fluorescence imaging, is employed as the output signal. Accompanied by digital imaging and automated counting analysis, E2 at the attomolar level can be measured. The smSimoa breaks the barrier of small molecule detection concentration and provides a basis for high throughput detection of multiple substances with fluorescence encoded magnetic beads.

Design and synthesis of DNA hydrogel based on EXPAR and CRISPR/Cas14a for ultrasensitive detection of creatine kinase MB.

Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas systems exhibit significant potential in developing biosensing technology due to their collateral cleavage capabilities. Herein, we introduced the collateral cleavage activity of CRISPR/Cas14a to activate DNA hydrogel for ultrasensitive detection of the myocardial infarction biomarker creatine kinase MB (CK-MB). In this strategy, the designed CRISPR/Cas14a system can be activated by introducing complementary DNA (cDNA) derived from competitive dissociation and exponential amplification (EXPAR), which is positively correlated with creatine kinase isoenzyme (CK-MB) concentration. Then the activated Cas14a protein can be utilized to indiscriminately cleave the DNA hydrogel cross-linker strand, leading to the degradation of the gel matrix and thus releasing the pre-encapsulated PtNPs/Cu-TCPP(Fe). PtNPs/Cu-TCPP(Fe) can trigger the TMB reaction, leading to an increase in absorbance value at 450 nm, thus enabling the quantitative detection of CK-MB. The proposed strategy combines CRISPR/Cas14a with DNA hydrogel for the first time, improving the programmability of DNA hydrogel and providing a reliable, sensitive, and versatile detection platform for trace non-nucleic acid targets.

CRISPR/dCas9-surface-enhanced Raman scattering for the detection of drug resistance gene macB.

Antibiotics have brought many benefits to public health systems worldwide since their first use in the last century, yet with their overuse in clinical treatment and livestock farming, new public health issues have arisen. Previously, we found in our experiments that the levels of macB genes in bovine raw milk ranked among the top of many drug resistance genes. In this paper, we present an analysis of regularly interspaced clustered short palindromic repeats (CRISPR) combined with surface-enhanced Raman scattering (SERS) technology for the detection of the drug resistance gene macB. The analysis was accomplished through the collaboration of the CRISPR system's ability to specifically identify genes and the more sensitive performance of the SERS. The analysis detects the drug resistance gene macB and does not yet require complex steps such as nucleic acid amplification. This method may prove to be an effective method for accurate detection of the drug-resistant gene macB, thus enabling more effective prevention of contamination of drug-resistant genes in food hygiene.

Quercetin improves rapid endothelialization and inflammatory microenvironment in electrospun vascular grafts.

There is a great need for small diameter vascular grafts among patients with cardiovascular diseases annually. However, continuous foreign body reactions and fibrosis capsules brought by biomaterials are both prone to poor vascular tissue regeneration. To address this problem, we fabricated a polycaprolactone (PCL) vascular graft incorporated with quercetin (PCL/QCT graft) in this study.In vitrocell assay showed that quercetin reduced the expressions of pro-inflammatory genes of macrophages while increased the expressions of anti-inflammatory genes. Furthermore,in vivoimplantation was performed in a rat abdominal aorta replacement model. Upon implantation, the grafts exhibited sustained quercetin release and effectively enhanced the regeneration of vascular tissue. The results revealed that quercetin improved endothelial layer formation along the lumen of the vascular grafts at four weeks. Furthermore, the thickness of vascular smooth muscle layers significantly increased in PCL/QCT group compared with PCL group. More importantly, the presence of quercetin stimulated the infiltration of a large amount of M2 phenotype macrophages into the grafts. Collectively, the above data reinforced our hypothesis that the incorporation of quercetin may be in favor of modulating the inflammatory microenvironment and improving vascular tissue regeneration and remodeling in vascular grafts.

Magnetic Halloysite Nanotube-Based SERS Biosensor Enhanced with Au@Ag Core-Shell Nanotags for Bisphenol A Determination.

Bisphenol A (BPA) has emerged as a contaminant of concern because long-term exposure may affect the human endocrine system. Herein, a novel aptamer sensor based on magnetic separation and surface-enhanced Raman scattering (SERS) is proposed for the extremely sensitive and specific detection of trace BPA. Moreover, the capture unit was prepared by immobilizing thiolated (SH)-BPA aptamer complementary DNA on AuNP-coated magnetic halloysite nanotubes (MNTs@AuNPs), and SH-BPA aptamer-modified Au@4-MBA@Ag core-shell SERS nanotags acted as signal units. By the complementary pairing of the BPA aptamer and the corresponding DNA, MNTs@AuNPs and Au@4-MBA@AgCS were linked together through hybridization-ligation, which acted as the SERS substrate. In the absence of BPA, the constructed aptamer sensor generated electromagnetic enhancement and plasmon coupling to improve the sensitivity of SERS substrates. Owing to the high affinity between BPA and the aptamer, the aptamer probe bound to BPA was separated from the capture unit by an externally-induced magnetic field. Thus, the Raman intensity of the MNTs@AuNP-Ag@AuCS core-satellite assemblies was negatively correlated with the BPA concentration. High sensitivity measurements of BPA might be performed by determining the decline in SERS signal strength together with concentration variations. The proposed aptasensor is a promising biosensing platform for BPA detection.

Amorphous poly-N-vinylcarbazole polymer as a novel matrix for the determination of low molecular weight compounds by MALDI-TOF MS.

Traditional matrices for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) are usually crystalline small molecules. The heterogeneous co-crystallization of the analyte and the matrix creates a sweet spot effect and reduces point-to-point reproducibility. In this study, an amorphous poly-N-vinylcarbazole polymer (PVK) was studied as a novel matrix for MALDI-TOF MS to detect various low molecular weight compounds (LMWCs) in the negative ion mode. The PVK achieved excellent matrix action and showed high sensitivity, good salt tolerance, and reproducibility. These results significantly broaden the design rules for new and efficient polymeric MALDI matrices.

Aptamer-based photonic crystals enable ultra-trace detection of staphylococcal enterotoxin B without labels.

Staphylococcal enterotoxin B (SEB) is one of the most common serotypes in staphylococcal food-poisoning cases. A rapid, sensitive, and simple method for SEB detection is crucial for public health. A photonic crystal (PC) sensing material for label-free detection of ultra-trace SEB was proposed in this study. Gold nanoparticle-doped silica microspheres were stacked to form an opal PC through self-assembly, and SEB aptamers, as the recognition element, were modified onto the PC. When the target protein of SEB came in contact with the PC sensing material, the reflection peak intensity of PCs decreased accordingly. The detection range was 1 × 10-6 to 1 ng mL-1, and the detection limit was 0.103 × 10-6 ng mL-1. Furthermore, the PC sensing material had great specificity and accuracy, which can be used for real sample monitoring. This PC sensing material achieved ultra-sensitive detection, which did not involve complicated preparation processes and reporter labelling.

A highly sensitive fluorometric biosensor for Fumonisin B1 detection based on upconversion nanoparticles-graphene oxide and catalytic hairpin assembly.

Fumonisin (FB) is a common mycotoxin in corn, wheat, oats, and their related products. FB1 is the most predominant among fumonisins and is responsible for severe food contamination that may have deleterious effects on public health. Therefore, the demand for achieving sensitive detection of FB1 is becoming more and more pressing. In the present study, a creative biosensor for FB1 detection was developed based on fluorescence resonance energy transfer between upconversion nanoparticles (UCNPs) and graphene oxide (GO) with catalytic hairpin assembly (CHA) target recycling and amplification. In the presence of FB1, UCNPs were bound to the CHA amplification products away from the GO surface. Simultaneously, a strong fluorescence signal was produced at 980 nm (near-infrared light). The correlation between the concentration of FB1 and the fluorescence intensity exhibited a high relevance (R2 = 0.9971) ranging from 0.032 to 500 ng mL-1, and the method reached a considerably low limit of detection (0.0121 ng mL-1). It could be applied for the detection of FB1 in corn, oats, and infant supplements. The sensitive detection of FB1 proves the application potential of this method on food safety detection. This biosensor can enable high-throughput fungal toxin detection by allowing the use of various aptamers.

Fabrication of Magnetic Al-Based Fe3O4@MIL-53 Metal Organic Framework for Capture of Multi-Pollutants Residue in Milk Followed by HPLC-UV.

The efficient capture of multi-pollutant residues in food is vital for food safety monitoring. In this study, in-situ-fabricated magnetic MIL-53(Al) metal organic frameworks (MOFs), with good magnetic responsiveness, were synthesized and applied for the magnetic solid-phase extraction (MSPE) of chloramphenicol, bisphenol A, estradiol, and diethylstilbestrol. Terephthalic acid (H2BDC) organic ligands were pre-coupled on the surface of amino-Fe3O4 composites (H2BDC@Fe3O4). Fe3O4@MIL-53(Al) MOF was fabricated by in-situ hydrothermal polymerization of H2BDC, Al (NO3)3, and H2BDC@Fe3O4. This approach highly increased the stability of the material. The magnetic Fe3O4@MIL-53(Al) MOF-based MSPE was combined with high-performance liquid chromatography-photo diode array detection, to establish a novel sensitive method for analyzing multi-pollutant residues in milk. This method showed good linear correlations, in the range of 0.05-5.00 µg/mL, with good reproducibility. The limit of detection was 0.004-0.108 µg/mL. The presented method was verified using a milk sample, spiked with four pollutants, which enabled high-throughput detection and the accuracies of 88.17-107.58% confirmed its applicability, in real sample analysis.