ABSTRACT
CRISPR-based diagnostics use the CRISPR-Cas system trans-cleavage activity to identify specific target sequences. When activated, this activity cleaves surrounding reporter molecules, producing a detectable signal. This technique has great specificity, sensitivity, and rapid detection, making it an important molecular diagnostic tool for medical and infectious disease applications. Despite its potential, the present CRISPR/Cas system has challenges with its single-stranded DNA reporters, characterized by low stability and limited sensitivity, restricting effective application in complex biological settings. In this work, we investigate the trans-cleavage activity of CRISPR/Cas12a on substrates utilizing fluorescent polystyrene microspheres to detect tetracycline. This innovative discovery led to the development of microsphere probes addressing the stability and sensitivity issues associated with CRISPR/Cas biosensing. By attaching the ssDNA reporter to polystyrene microspheres, we discovered that the Cas12a system exhibits robust and sensitive trans-cleavage activity. Further work revealed that the trans-cleavage activity of Cas12a on the microsphere surface is significantly dependent on the concentration of the ssDNA reporters. Building on these intriguing discoveries, we developed microsphere-based fluorescent probes for CRISPR/Cas aptasensors, which showed stability and sensitivity in tetracycline biosensing. We demonstrated a highly sensitive detection of tetracycline with a detection limit of 0.1 µM. Finally, the practical use of a microsphere-based CRISPR/Cas aptasensor in spiked food samples was proven successful. These findings highlighted the remarkable potential of microsphere-based CRISPR/Cas aptasensors for biological research and medical diagnosis.
Subject(s)
Biosensing Techniques , CRISPR-Cas Systems , DNA, Single-Stranded , Microspheres , Polystyrenes , Tetracycline , Tetracycline/analysis , CRISPR-Cas Systems/genetics , Polystyrenes/chemistry , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/genetics , Biosensing Techniques/methods , Aptamers, Nucleotide/chemistry , Aptamers, Nucleotide/metabolism , CRISPR-Associated Proteins/genetics , CRISPR-Associated Proteins/metabolism , Fluorescent Dyes/chemistry , Bacterial Proteins , EndodeoxyribonucleasesABSTRACT
The country bean (Lablab purpureus), is a significant contributor of dietary protein and other essential components in human nutrition. Because of its elevated moisture content, it is susceptible to rapid decay, leading to losses after harvesting. The utilization of solar drying has attracted significant attention as a tactic to minimize nutrient depletion in dried goods and enhance their longevity. This study employed four solar drying techniques, namely long chimney, short chimney, box solar drying and open sun drying, along with pretreatments such as potassium metabisulfite, potassium-sodium tartrate, citric acid and ascorbic acid. The objective was to determine an effective solar drying method, combined with pretreatment, that can maintain the color and nutritional qualities of dried country bean seeds. The treatment combinations were organized in a factorial randomized complete block design (RCBD) with three replications. The data were subjected to a two-way analysis of variance (ANOVA) and a Duncan Multiple Range Test (DMRT) was conducted at a significance level of 5 % (p < 0.05). Results revealed that box solar dryer having the highest drying efficiency, retained the highest ß-carotene (82.94 %), vitamin C (90.15 %), protein (96.48 %), fat (11.63 %), and ash (90.50 %) with maximum DPPH radical scavenging activity (lowest IC50 209.49 µg/ml) compared to other driers. Besides, country bean seeds have noteworthy proximate compositions, antioxidant activity, and bioactive components treated with 1 % potassium metabisulfite. Furthermore, the country bean seeds dehydrated in box solar dryer after 1 % potassium metabisulfite treatment received the highest acceptance score on the five-point Hedonic scale (4.83-4.89 out of 5.00) and color appearance and the similar trend was further supported by principal component analysis. Thus, it can be inferred that using a box solar dryer with a 1 % potassium metabisulfite pretreatment is a feasible method for preserving the color and nutritional value of country bean seeds and reducing postharvest losses.
ABSTRACT
Microbial-assisted rhizoengineering is a promising biotechnology for improving crop productivity. In this study, lettuce roots were bacterized with two lead (Pb) tolerant rhizobacteria including Pseudomonas azotoformans ESR4 and P. poae ESR6, and a consortium consisted of ESR4 and ESR6 to increase productivity, physiology and antioxidants, and reduce Pb accumulation grown in Pb-contaminated soil i.e., 80 (Pb in native soil), 400 and 800 mg kg-1 Pb. In vitro studies showed that these strains and the consortium produced biofilms, synthesized indole-3-acetic acid and NH3, and solubilized phosphate challenging to 0, 100, 200 and 400 mg L-1 of Pb. In static conditions and 400 mg L-1 Pb, ESR4, ESR6 and the consortium adsorbed 317.0, 339.5 and 357.4 mg L-1 Pb, respectively, while 384.7, 380.7 and 373.2 mg L-1 Pb, respectively, in shaking conditions. Fourier transform infrared spectroscopy results revealed that several functional groups [Pb-S, M - O, O-M-O (M = metal ions), S-S, PO, CO, -NH, -NH2, C-C-O, and C-H] were involved in Pb adsorption. ESR4, ESR6 and the consortium-assisted rhizoengineering (i) increased leaf numbers and biomass production, (ii) reduced H2O2 production, malondialdehyde, electrolyte leakages, and transpiration rate, (iii) augmented photosynthetic pigments, photosynthetic rate, water use efficiency, total antioxidant capacity, total flavonoid content, total phenolic content, and minerals like Ca2+ and Mg2+ in comparison to non-rhizoengineering plants grown in Pb-contaminated soil. Principal component analysis revealed that higher pigment production and photosynthetic rate, improved water use efficiency and increased uptake of Ca2+ were interlinked to increased productivity by bacterial rhizoengineering of lettuce grown in different levels of Pb exposures. Surprisingly, Pb accumulation in lettuce roots and shoots was remarkably decreased by rhizoengineering than in non-rhizoengineering. Thus, these bacterial strains and this consortium could be utilized to improve productivity and reduce Pb accumulation in lettuce.
Subject(s)
Lactuca , Lead , Plant Roots , Soil Pollutants , Lactuca/growth & development , Lactuca/metabolism , Lead/metabolism , Soil Pollutants/metabolism , Plant Roots/metabolism , Plant Roots/microbiology , Soil Microbiology , Biodegradation, Environmental , Soil/chemistry , Pseudomonas/metabolism , Pseudomonas/physiology , Antioxidants/metabolismABSTRACT
In this study, a nanocomposite was synthesized by incorporating graphitic carbon nanosheets, carboxyl-functionalized multiwalled carbon nanotubes, and zirconium oxide nanoparticles. The resulting nanocomposite was utilized for the modification of a glassy carbon electrode. Subsequently, matrix metalloproteinase aptamer (AptMMP-9) was immobilized onto the electrode surface through the application of ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride-N-hydroxysuccinimide (EDC-NHS) chemistry. Morphological characterization of the nanomaterials and the nanocomposite was performed using field-emission scanning electron microscopy (FESEM). The nanocomposite substantially increased the electroactive surface area by 205%, facilitating enhanced immobilization of AptMMP-9. The efficacy of the biosensor was evaluated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Under optimal conditions, the fabricated sensor demonstrated a broad range of detection from 50 to 1250 pg/mL with an impressive lower limit of detection of 10.51 pg/mL. In addition, the aptasensor exhibited remarkable sensitivity, stability, excellent selectivity, reproducibility, and real-world applicability when tested with human serum and saliva samples. In summary, our developed aptasensor exhibits significant potential as an advanced biosensing tool for the point-of-care quantification of MMP-9, promising advancements in biomarker detection for practical applications.
Subject(s)
Aptamers, Nucleotide , Graphite , Nanocomposites , Nanotubes, Carbon , Nitrogen Compounds , Zirconium , Humans , Nanotubes, Carbon/chemistry , Matrix Metalloproteinase 9 , Electrochemical Techniques/methods , Saliva , Reproducibility of Results , Limit of Detection , Aptamers, Nucleotide/chemistry , Nanocomposites/chemistryABSTRACT
Discharge of untreated dyeing wastewater nearby water-bodies is one of major causes of water pollution. Generally, bacterial strains isolated from industrial effluents and/or contaminated soils are used for the bioremediation of Methyl orange (MO), a mutagenic recalcitrant mono-azo dye, used in textiles and biomedical. However, MO degradation by biofilm producing plant growth-promoting rhizobacteria (BPPGPR) was not studied yet. In this study, 19 out of 21 BPPGPR strains decolorized 96.3-99.9% and 89.5-96.3% MO under microaerophilic and aerobic conditions, respectively from Luria-Bertani broth (LBB) followed by yeast-extract peptone and salt-optimized broth plus glycerol media within 120 h of incubation at 28 °C. Only selected BPPGPR including Pseudomonas fluorescens ESR7, P. veronii ESR13, Stenotrophomonas maltophilia ESR20, Staphylococcus saprophyticus ESD8, and P. parafulva ESB18 were examined for process optimization of MO decolorization using a single factor optimization method. This study showed that under optimal conditions (e.g., LBB, 100 mg L-1 MO, pH 7, incubation of 96 h, 28 °C), these strains could remove 99.1-99.8% and 97.6-99.5% MO under microaerophilic and aerobic conditions, respectively. Total azoreductase and laccase activities responsible for biodegradation were also remarkably activated in the biodegraded samples under optimal conditions, while these activities were repressed under unfavorable conditions (e.g., 40 °C and 7.5% NaCl). This study confirmed that MO was degraded and detoxified by these bacterial strains through breakage of azo bond. So far, this is the first report on bioremediation of MO by the BPPGPR strains. These BPPGPR strains are highly promising to be utilized for the bioremediation of dyeing wastewater in future.
Subject(s)
Coloring Agents , Wastewater , Coloring Agents/chemistry , Mutagens , Biodegradation, Environmental , Bacteria/metabolism , Azo Compounds/chemistryABSTRACT
Enzyme mimicking nanomaterials, nanozymes, have gained considerable interest in the scientific community because of their superior properties compared to natural enzymes, including their high stability at extreme conditions, cheaper availability, and ease of synthesis. Herein, we report novel colloidal gold nanoparticles - graphene nanoplatelets - chitosan (CS) with peroxidase mimicking properties used to carry out highly sensitive and selective immunoassay for porcine gelatin detection. The interaction between anti-gelatin antibody conjugated nanozyme with porcine gelatin proteins produced an ultrasensitive immunoassay response in the form of a colourimetric signal directly proportional to the porcine gelatin protein concentration. The nanozyme produced a colourimetric response in the presence of its substrate, 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2), demonstrating its peroxidase mimicking properties. The results revealed that the nanozyme exhibited remarkable selectivity and sensitivity in the assay, detecting proteins at concentrations as low as 86.42 pg/mL. Additionally, the immunosensor demonstrated a broad linear detection range spanning from 200 pg/mL to 2 ng/mL.
Subject(s)
Biosensing Techniques , Metal Nanoparticles , Swine , Metal Nanoparticles/chemistry , Gelatin , Hydrogen Peroxide/chemistry , Colorimetry/methods , Biosensing Techniques/methods , Gold , Immunoassay/methods , Peroxidase/chemistry , AnimalsABSTRACT
A versatile and highly sensitive sensing platform based on nanocellulose/MXene/ZrO2 nano framework has been developed at the surface of a glassy carbon electrode (GCE) for detecting ovalbumin (Ova). To create this innovative nano framework, dialdehyde groups were introduced onto the surface of cellulose nanofibers (CNFs), which were then decorated with MXene nanosheets and nanostructured zirconia. Nanocellulose/MXene/ZrO2 nano framework was used as electrochemical mediator and immobilization environment that provided the large surface area and 197 % increment in the electrochemical signal which allowed the Ova detection in the femtomolar range. Ovalbumin antibody was immobilized on the surface of dialdehyde cellulose nanofiber through covalent bonding between amino groups of Ova and dialdehyde groups of CNFs. The assembly process of nano framework, anti-Ova, and Ova antigen were characterized using electrochemical approaches (CV and DPV). The fabricated immunosensor is further applied to DPV detection of Ova and it demonstrated a linear response to Ova antigen in the linear range of 0.01-1000 pg/mL. With optimal experimental conditions, the detection limit, quantification limit and sensitivity of Ova were found to be 1.1 fg/mL, 0.01 pg/mL and 0.1497 µA pg/mL cm-2, respectively. The fabricated immunosensor exhibited high selectivity, reproducibility, and interference resistance and achieved excellent recoveries in real food samples spiked with Ova, indicating its potential applicability in food safety monitoring.
Subject(s)
Biosensing Techniques , Nitrites , Transition Elements , Ovalbumin , Immunoassay , Reproducibility of Results , Titanium , Electrochemical Techniques , Limit of Detection , Gold/chemistryABSTRACT
This study reports a novel biosensing system that leverages recombinase polymerase amplification (RPA) in conjunction with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a technology, integrated with a nanozyme (NZ) based on cerium dioxide (CeO2). With the integration of CeO2 NZ, a dual-mode detection platform could be developed for Salmonella detection using fluorometric and colourimetric assays. The CRISPR/Cas12a system, when activated in the presence of target DNA, could cleave the FAM-labelled probe to lead to a fluorometric response. Also, when the CeO2 NZ was introduced in the presence of H2O2, a colourimetric response was generated, directly proportional to the concentration of target DNA present. We hypothesise that adding highly reactive H2O2 within the post-CRISPR/Cas12a reaction system allows for increased release of hydroxyl free radicals within the mixture. Thus, the double recognition through NZ and the CRISPR/Cas12a system provided enhanced selectivity and sensitivity to the method. The proposed biosensor could successfully detect Salmonella at concentrations as low as 0.88 pg/µL and 1.28 pg/µL for fluorometric and colourimetric responses, respectively. Furthermore, the developed biosensor could be applied in real sample analysis of raw food samples (chicken, egg, and beef) to give a good recovery in the spiked food samples with varying concentrations of cultured bacterial DNA.
Subject(s)
Biosensing Techniques , Recombinases , Cattle , Animals , CRISPR-Cas Systems/genetics , Hydrogen Peroxide , DNA, BacterialABSTRACT
We propose a CRISPR/Cas12a-mediated recombinase polymerase amplification (RPA) detection method that combines RPA with Cas12a cleavage for the detection of halal food adulteration, which is of global concern, particularly for Muslim consumers. We optimized the reagent concentrations for the Cas12a cleavage steps and designed and screened gRNA targeting a conserved area of the mitochondrial cytochrome C oxidase subunit I (COX1) gene. This procedure successfully detected the presence of porcine components as low as 5 pg/µL in the linear range of 5-1000 pg/µL. The assay's detection limit was 500 times lower than CRISPR-based approaches that exclude a preamplification step, allowing the detection of trace porcine DNA in food samples. The assay additionally showed no cross-reaction with nontarget species. Therefore, this detection platform shows tremendous potential as a method for the quick, sensitive, and specific detection of porcine-derived components.
ABSTRACT
This study introduces CRISPR/Cas-based aptasensor for the highly sensitive and specific detection of the antibiotic, ampicillin. Ampicillin (AMPI) is a commonly used antibiotic for treating pathogenic bacteria and is additionally added to livestock feed in agriculture. This study can enable early detection of antibiotic residues, prevent their accumulation in the environment, and ensure compliance with food safety regulations. Herein, the aptasensor was developed with the CRISPR/Cas system by utilizing three different ampicillin-specific aptamers, each conjugated with a biotin at the 5'-end. The ssDNA activator was bound to the aptamers through complementary base pairings. The attraction of the aptamers to the ampicillin target released the bound ssDNA, causing the activation of the CRISPR/Cas system. The DNA reporter probe, labelled with Cy3 and a quencher, turns on the fluorescence signal when cleaved by the activated Cas12a through trans-cleavage measured using a fluorescence spectrophotometer at 590 nm. The fluorescence signal was linearly proportional to the ampicillin target concentration with a 0.01 nM limit of detection and a read-out time of 30 min. This aptasensor showed high sensitivity towards ampicillin even in the presence of other antibiotics. The method was also successfully implemented for ampicillin detection in spiked food samples.
Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , CRISPR-Cas Systems/genetics , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Ampicillin , Anti-Bacterial Agents/pharmacology , DNA, Single-StrandedABSTRACT
This review highlights the recent advancements in the field of nanozymes and their applications in the development of point-of-care biosensors. The use of nanozymes as enzyme-mimicking components in biosensing systems has led to improved performance and miniaturization of these sensors. The unique properties of nanozymes, such as high stability, robustness, and surface tunability, make them an attractive alternative to traditional enzymes in biosensing applications. Researchers have explored a wide range of nanomaterials, including metals, metal oxides, and metal-organic frameworks, for the development of nanozyme-based biosensors. Different sensing strategies, such as colorimetric, fluorescent, electrochemical and SERS, have been implemented using nanozymes as signal-producing components. Despite the numerous advantages, there are also challenges associated with nanozyme-based biosensors, including stability and specificity, which need to be addressed for their wider applications. The future of nanozyme-based biosensors looks promising, with the potential to bring a paradigm shift in biomolecular sensing. The development of highly specific, multi-enzyme mimicking nanozymes could lead to the creation of highly sensitive and low-biofouling biosensors. Integration of nanozymes into point-of-care diagnostics promises to revolutionize healthcare by improving patient outcomes and reducing costs while enhancing the accuracy and sensitivity of diagnostic tools.
Subject(s)
Biosensing Techniques , Metal-Organic Frameworks , Nanostructures , Humans , Nanostructures/chemistry , Metals , Oxides , Metal-Organic Frameworks/chemistry , CatalysisABSTRACT
For early detection and diagnosis of cancer, it is essential to develop an electrochemical biosensor that is quick, accurate, and sensitive. Here, we use gold nanorod (AuNR) and gold nanowire (AuNW) nanocomposites (AuNR/AuNW/CS) as electrode modifiers on a glassy carbon electrode (GCE) to construct a sensitive label-free electrochemical aptasensor to detect ApoA1. The thiolated ApoA1-specific aptamers were immobilized onto the modified electrode surface through self-assembled monolayers. Electrochemical techniques, such as cyclic voltammetry (CV) and differential pulse voltammetry (DPV), were used to analyze the fabrication steps. The concentration of ApoA1 was measured with DPV on the aptasensor, with a linear range of 0.1 to 1000 pg mL-1 and a detection limit of 0.04 pg mL-1. When compared to results from ELISA tests (which have a detection limit of 80 pg mL-1), the results achieved here were over 2000 times better. The aptasensor's performance was successfully evaluated using human serum spiked with ApoA1, suggesting that it has great potential for practical application. The electrochemical apatsensor additionally demonstrated outstanding selectivity responses and strong stability toward the target analyte.
ABSTRACT
The tropical estuarine ecosystem is fascinating for studying the dynamics of water quality and phytoplankton diversity due to its frequently changing hydrological conditions. Most importantly, phytoplankton is the main supplier of ω3 polyunsaturated fatty acids (PUFA) in the coastal food web for fish as they could not synthesize PUFA. This study evaluated seasonal variations of water quality parameters in the Meghna River estuary (MRE), explored how phytoplankton diversity changes according to hydro-chemical parameters, and identified the major phytoplankton groups as the main source of PUFA for hilsa fish. Ten water quality indicators including temperature, dissolved oxygen, pH, salinity, dissolved inorganic nitrogen (DIN = nitrate, nitrite, ammonia) and phosphorus, dissolved silica and chlorophyll-a were evaluated. In addition, phytoplankton diversity was assessed in the water and hilsa fish gut. Principal component analysis (PCA) was used to analyze the spatio-temporal changes in the water quality conditions, and the driving factors in the MRE. Four main components were extracted and explained 75.4% variability of water quality parameters. The most relevant driving factors were dissolved oxygen, salinity, temperature, and DIN (nitrate, nitrite and ammonia). These variabilities in physicochemical parameters and dissolved inorganic nutrients caused seasonal variations in two major groups of phytoplankton. Peak abundance of Chlorophyta (green algae) occurred in water in nutrient-rich environments (nitrogen and phosphorus) during the wet (36%) season, while Bacillariophyta (diatoms) were dominant during the dry (32%) season that depleted dissolved silica. Thus, the decrease of green algae and the increase of diatoms in the dry season indicated the potential link to seasonal changes of hydro-chemical parameters. The green algae (53.7%) were the dominant phytoplankton group in the hilsa gut content followed by diatoms (22.6%) and both are contributing as the major source of PUFAs for hilsa fish according to the electivity index as they contain the highest amounts of PUFAs (60 and 28% respectively).
Subject(s)
Chlorophyta , Diatoms , Animals , Phytoplankton , Estuaries , Ecosystem , Nitrates/analysis , Nitrites/analysis , Ammonia/analysis , Bangladesh , Environmental Monitoring , Phosphorus/analysis , Seasons , Nitrogen/analysis , Fishes , Oxygen/analysis , Fatty Acids, Unsaturated , Silicon Dioxide/analysisABSTRACT
The Pasur River estuary (PRE), the largest estuary in the Sundarbans mangrove area, provides vital fishery resources and supports millions of livelihoods in the southwestern coastal region of Bangladesh. This study focused on the tidal and run-off effects on the outwelling of nutrients from the Sundarbans mangrove creeks to the PRE. Spatial and temporal variations of nutrient and chlorophyll-a concentrations were assessed by water sampling at 11 stations in the study area from January to December 2019. Dissolved inorganic nutrients and chlorophyll-a were analyzed by standard methods using a spectrophotometer. In the tidal mangrove creeks, dissolved inorganic nitrogen, phosphate, and silica concentrations were significantly higher (p < 0.05) during the spring tide than those during the neap tide, suggesting that these nutrients were flushed from the mangrove area by the inundation and tidal mixing of the spring tide. In general, chlorophyll-a (mean ± SD) concentrations in the PRE and the tidal mangrove creeks were 5.62 ± 1.30 µg/L and 9.03 ± 0.59 µg/L in the wet season, respectively. During the dry season, the chlorophyll-a decreased to 4.37 µg/L ± 0.68 and 4.94 ± 1.52 µg/L in the PRE and the tidal mangrove creek, respectively. The amount of nutrients outwelled from the mangrove creeks to the estuary was 1.53 ± 0.67 mg/L DIP, 0.001 ± 0.0004 mg/L DIN, and 1.38 ± 0.48 mg/L dissolved silica. DIP, silica, and chlorophyll-a concentrations were significantly higher (p < 0.05) during the spring tide compared to the neap tide, but salinity was not significantly (p > 0.05) different between the two tidal levels. This study showed that the mangrove creeks formed an important link in transporting nutrients from the mangrove forest to the estuary.
ABSTRACT
A molybdenum sulfide/zirconium oxide/Nafion (MoS2/ZrO2/Naf) based electrochemiluminescence (ECL) aptasensor for the selective and ultrasensitive detection of ApoA1 is proposed, with Ru(bpy)3 2+ as the luminophore. The chitosan (CS) modification on the nanocomposite layer allowed glutaraldehyde (GLUT) cross-linking, resulting in the immobilization of ApoA1 aptamers. Scanning electron microscopy, tunneling electron microscopy, and energy dispersive X-ray spectroscopy were used to characterize the nanocomposite, while electrochemiluminescence (ECL), cyclic voltammetry, and electrochemical impedance spectroscopy were used to analyze the aptasensor assembly. The nanocomposite was used as an electrode modifier, which increased the intensity of the ECL signal. Due to the anionic environment produced on the sensor surface following the specific interaction of the ApoA1 biomarker with the sensor, more Ru(bpy)3 2+ were able to be electrostatically attached to the aptamer-ApoA1 complex, resulting in enhanced ECL signal. The ECL aptasensor demonstrated outstanding sensitivity for ApoA1 under optimal experimental conditions, with a detection limit of 53 fg/mL and a wide linear dynamic range of 0.1-1000 pg/mL. The potential practical applicability of this aptasensor was validated by analyzing ApoA1 in human serum samples, with recovery rates of 94-108% (n = 3). The proposed assay was found to be substantially better compared to the commercially available enzyme-linked immunosorbent assay method, as reflected from over 1500 times improvement in the detection limit for ApoA1.
ABSTRACT
Nanozymes have piqued the curiosity of scientists in recent years because of their ability to demonstrate enzyme-like activity combined with advantages such as high stability, inexpensive availability, robust activity, and tunable properties. These attributes have allowed the successful application of nanozymes in sensing to detect various chemical and biological target analytes, overcoming the shortcomings of conventional detection techniques. In this review, we discuss recent developments of nanozyme-based sensors to detect biomarkers associated with food quality and safety. First, we present a brief introduction to this topic, followed by discussing the different types of sensors used in food biomarker detection. We then highlight recent studies on nanozyme-based sensors to detect food markers such as toxins, pathogens, antibiotics, growth hormones, metal ions, additives, small molecules, and drug residues. In the subsequent section, we discuss the challenges and possible solutions towards the development of nanozyme-based sensors for application in the food industry. Finally, we conclude the review by discussing future perspectives of this field towards successful detection and monitoring of food analytes.
ABSTRACT
In recent years, rapid detection methods such as polymerase chain reaction (PCR) and quantitative real-time PCR (qPCR) have been continuously developed to improve the detection of food-borne pathogens in food samples. The recent developments of PCR and qPCR in the detection and identification of these food-borne pathogens are described and elaborated throughout this review. Specifically, further developments and improvements of qPCR are discussed in detecting Salmonella and norovirus. Promising advances in these molecular detection methods have been widely used to prevent human food-borne illnesses and death caused by the food-borne pathogens. In addition, this review presents the limitations and challenges of the detection methods which include conventional culture method and conventional PCR method in detecting Salmonella and norovirus. Furthermore, several advances of qPCR such as viability PCR (vPCR) and digital PCR (dPCR) have been discussed in the detection of Salmonella and norovirus. Good practice of analysis of the food-borne pathogens and other contaminants in the food industry as well as the advancement of molecular detection methods will help improve and ensure food safety and food quality.
ABSTRACT
Biosensor technology is experiencing an unprecedented momentum that is driven by rapid advances in material science for utilization in various fields, and is constantly evolving to develop fast, simple and sensitive sensors. Here, we present a comprehensive and concise review on electrochemiluminescent (ECL) nanomaterial-based aptasensors including challenges and applications. ECL aptasensor combines the merits of a biosensor, selectivity of an aptamer, and sensitivity of ECL to construct sensitive and selective sensors. Signal amplification processes through the incorporation of nanomaterials are essential to further enhance the sensitivity of ECL sensing systems. Nanomaterials are commonly utilized in sensor fabrication as (i) electrode modifiers, (ii) nanocarriers, (iii) nanocatalysts, and (iv) luminophores. The review highlights the effects of integrating nanomaterials such as gold nanoparticles, metal nanoparticles, metal oxides, carbon nanomaterials, metal organic frameworks, and quantum dots using various research strategies that address signal amplification to enhance the ECL signal. Furthermore, various concepts have been demonstrated with relevant figures and explanations to serve as a general overview for the design of ECL aptasensors. Lastly, we discuss the underlying problems and possibilities for ECL aptasensor research and development.
Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Metal Nanoparticles , Nanostructures , Electrochemical Techniques , Gold , Luminescent MeasurementsABSTRACT
This work reports on the first electrochemiluminescence (ECL) immunosensor employing a novel nanostructured composite of titanium dioxide (TiO2) and gold nanourchins (AuNU) to detect protein allergen Ovomucoid (Ovm) found in eggs. TiO2 and AuNU were dispersed in Nafion and drop-casted onto SPGE with Tris(2,2'-bipyridyl)-ruthenium (II) ([Ru(bpy)3]2+) and tri-n-propylamine (TPrA) served as strong luminophore/co-reactant pairs as a source of ECL signals. The linear range, limit of detection, reproducibility and practical applications of the sensor were assessed. The fabricated ECL immuosensor produced a promising limit of detection of as low as 0.01 pg/mL. Two linear ranges of 0.01-50 pg/mL and 100-750 pg/mL with corresponding correlation coefficients of R2 = 0.99136 and R2 = 0.97829) respectively, were determined. Despite its simple fabrication method, this label-free immunosensor also showcased excellent selectivity, reproducibility, interference-resistance and yielded outstanding recoveries between 97.42 and 104.05% of Ovm analysis in spiked real food samples.
Subject(s)
Biosensing Techniques , Nanocomposites , Biosensing Techniques/methods , Electrochemical Techniques/methods , Fluorocarbon Polymers , Gold , Immunoassay/methods , Limit of Detection , Luminescent Measurements/methods , Ovomucin , Reproducibility of Results , TitaniumABSTRACT
Energy harvesting and storage is highly demanded to enhance the lifetime of autonomous systems, such as IoT sensor nodes, avoiding costly and time-consuming battery replacement. However, cost efficient and small-scale energy harvesting systems with reasonable power output are still subjects of current development. In this work, we present a mechanically and magnetically excitable MEMS vibrational piezoelectric energy harvester featuring wafer-level integrated rare-earth micromagnets. The latter enable harvesting of energy efficiently both in resonance and from low-g, low-frequency mechanical energy sources. Under rotational magnetic excitation at frequencies below 50 Hz, RMS power output up to 74.11 µW is demonstrated in frequency up-conversion. Magnetic excitation in resonance results in open-circuit voltages > 9 V and RMS power output up to 139.39 µW. For purely mechanical excitation, the powder-based integration process allows the realization of high-density and thus compact proof masses in the cantilever design. Accordingly, the device achieves 24.75 µW power output under mechanical excitation of 0.75 g at resonance. The ability to load a capacitance of 2.8 µF at 2.5 V within 30 s is demonstrated, facilitating a custom design low-power ASIC.