Electrochemical Nanoaptasensor Based on Graphitic Carbon Nitride/Zirconium Dioxide/Multiwalled Carbon Nanotubes for Matrix Metalloproteinase-9 in Human Serum and Saliva.

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.

A novel and ultrasensitive electrochemical immunosensor based on nanocellulose-Ti3C2Tx@ZrO2 nano framework for the detection of ovalbumin.

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.

A CRISPR/Cas12a-based fluorescence aptasensor for the rapid and sensitive detection of ampicillin.

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.

An electrochemical aptasensor based on AuNRs/AuNWs for sensitive detection of apolipoprotein A-1 (ApoA1) from human serum.

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.

Nanozyme-based sensors for detection of food biomarkers: a review.

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.

A solid-state electrochemiluminescence aptasensor for ß-lactoglobulin using Ru-AuNP/GNP/Naf nanocomposite-modified printed sensor.

An electrochemiluminescence (ECL) aptasensor for the detection of the milk protein allergen ß-lactoglobulin (ß-LG) using nanocomposite as luminophore was fabricated. The Ru-AuNPs/GNP/Naf complex was formed by combining the Rubpy32+-AuNPs complex (Ru-AuNPs), prepared by modifying the negatively charged surface of gold nanoparticles (AuNPs) with positively charged Rubpy32+ through electrostatic interactions and the graphene nanoplatelets-Nafion (GNP/Naf) at a ratio of 2:1. The nanocomposite was coated on the surface of the screen-printed electrode (SPCE) through the film-forming properties of Nafion. A layer of chitosan (CS) was coated onto this modified electrode, and later amine-terminated ß-LG aptamers were covalently attached to the CS/Ru-AuNP/GNP/Naf via glutaraldehyde (GLUT) cross-linking. When ß-LG was incubated with the aptasensor, a subsequent decrease in ECL intensity was recorded. Under the optimal conditions, the ECL intensity of the aptasensor changed linearly with the logarithmic concentration of ß-LG, in the range 0.1 to 1000 pg/ml, and the detection limit was 0.02 pg/mL (3σ/m). The constructed aptasensor displayed simple and fast determination of ß-LG with excellent reproducibility, stability, and high specificity. Additionally, the proposed ECL aptasensor displayed high recoveries (92.5-112%) and low coefficients of variation (1.6-7.8%), when ß-LG fortified samples were analyzed. Integrating Ru-AuNPs/GNP/Naf nanocomposite in the ECL aptasensor paves the way towards a cost-effective and sensitive detection of the milk allergen ß-LG.

Recent trends in nanomaterial-based signal amplification in electrochemical aptasensors.

Ultrasensitive biosensors have become a necessity in the world of scientific research, and several signal enhancement strategies have been employed to attain exceptionally low detection limits. Nanotechnology turns out to be a strong contender for signal amplification, as they can be employed as platform modifiers, catalysts, carriers or labels. Here, we have described the most recent advancements in the utilization of nanomaterials as signal amplification components in aptamer-based electrochemical biosensors. We have briefly reviewed the methods that utilized nanomaterials, namely gold and carbon, as well as nanocomposites such as: graphene, carbon nanotubes, quantum dots, and metal-organic frameworks.

A Highly Sensitive Label-free Aptasensor Based on Gold Nanourchins and Carbon Nanohorns for the Detection of Lipocalin-2 (LCN-2).

A synergistic nanocomposite film composed of gold nanourchins (AuNU), oxidised carbon nanohorns (CNH), and chitosan functioned as an electrode modifier in the fabrication of the sensitive lipocalin-2 (LCN-2) aptasensor. The AuNUs/CNH/CS composite increased the surface area and thereby amplified the signal transduction. The amine-terminated LCN-2 aptamer was immobilised through the amide bond formed between the carboxyl group of polyglutamic acid (PGA) and the amine group of aptamer. Interaction of LCN-2 with the aptamer caused conformational changes in the structure of the aptamer. This generated higher conductivity, resulting in increased DPV peak current. The DPV signal increased with increasing concentration of LCN-2, and the change in signal was used for quantitative detection. The proposed aptasensor was able to detect LCN-2 in the linear range of 0.1 - 100.0 pg mL-1, with a low detection limit of 10 fg mL-1. The aptasensor showed high sensitivity, selectivity, reproducibility, and was able to detect LCN-2 in serum samples.

Electrochemiluminescence immunosensor for tropomyosin using carbon nanohorns/Nafion/Fe3O4@Pd screen-printed electrodes.

This study focuses on developing a highly sensitive immunosensor by immobilizing oxidized carbon nanohorns/Nafion/Fe3O4@Pd nanocomposite on carbon screen-printed electrodes (SPEs) for the detection of tropomyosin (Tro-Ag). The performance of the fabricated immunosensor was investigated via electrochemiluminescence (ECL) method, resulting from the chemical reaction between tris(bipyridine)ruthenium(II) chloride ([Ru(bpy)3]Cl2) and tripropylamine (TPrA), and the peak intensity is recorded at 1.0 V. The nanocomposite is able to enhance the ECL intensity of [Ru(bpy)3]2 +/TPrA system and achieves high sensitivity of 28.16 fg/mL with a dynamic working range of 28.16 fg/mL to 100 ng/ml.. Furthermore, the immunosensor demonstrated a decent stability and good repeatability for Tro-Ag detection in food products. Graphical abstract The schematic representation of the modified carbon SPE with CNHs-OH/Nafion/Fe3O4@Pd and the signal produced in the (a) absence and (b) presence of Tro-Ag.

Trends and Advances in Electrochemiluminescence Nanobiosensors.

The rapid and increasing use of the nanomaterials (NMs), nanostructured materials (NSMs), metal nanoclusters (MNCs) or nanocomposites (NCs) in the development of electrochemiluminescence (ECL) nanobiosensors is a significant area of study for its massive potential in the practical application of nanobiosensor fabrication. Recently, NMs or NSMs (such as AuNPs, AgNPs, Fe3O4, CdS QDs, OMCs, graphene, CNTs and fullerenes) or MNCs (such as Au, Ag, and Pt) or NCs of both metallic and non-metallic origin are being employed for various purposes in the construction of biosensors. In this review, we have selected recently published articles (from 2014-2017) on the current development and prospects of label-free or direct ECL nanobiosensors that incorporate NCs, NMs, NSMs or MNCs.

Trends in Paper-based Electrochemical Biosensors: From Design to Application.

Electrochemical bio-sensing using paper-based detection systems is the main focus of this review. The different existing designs of 2-dimensional and 3-dimensional sensors, and fabrication techniques are discussed. This review highlights the effect of adopting different sensor designs, distinct fabrication techniques, as well as different modification methods, in order to produce reliable and reproducible reading. The use of various nanomaterials have been demonstrated in order to modify the surface of electrodes during fabrication to further enhance the signal for subsequent analysis. The reviewed sensors were classified into categories based on their applications, such as diagnostics, environmental and food testing. One of the major advantages of using paper-based electrochemical sensors is the potential for miniaturization, which only requires relatively small amount of samples, and the low cost for the purpose of mass production. Additionally, most of the devices reviewed were made to be portable, making them well-suited for on-site detection. Finally, paper-based detection is an ideal platform to fabricate cost-effective, user-friendly and sensitive electrochemical biosensors, with large capacity for customization depending on functional needs.

Colorimetric Nucleic Acid Detection on Paper Microchip Using Loop Mediated Isothermal Amplification and Crystal Violet Dye.

Nucleic acid detection is of paramount importance in monitoring of microbial pathogens in food safety and infectious disease diagnostic applications. To address these challenges, a rapid, cost-effective label-free technique for nucleic acid detection with minimal instrumentations is highly desired. Here, we present paper microchip to detect and quantify nucleic acid using colorimetric sensing modality. The extracted DNA from food samples of meat as well as microbial pathogens was amplified utilizing loop-mediated isothermal amplification (LAMP). LAMP amplicon was then detected and quantified on a paper microchip fabricated in a cellulose paper and a small wax chamber utilizing crystal violet dye. The affinity of crystal violet dye toward dsDNA and positive signal were identified by changing the color from colorless to purple. Using this method, detection of Sus scrofa (porcine) and Bacillus subtilis (bacteria) DNA was possible at concentrations as low as 1 pg/µL (3.43 × 10 -1 copies/µL) and 10 pg/µL (2.2 × 103 copies/µL), respectively. This strategy can be adapted for detection of other DNA samples, with potential for development of a new breed of simple and inexpensive paper microchip at the point-of-need.