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The early diagnosis of diseases is of great importance for the effective treatment of patients. Biomarkers are one of the most promising medical approaches in the diagnosis of diseases and their progress and facilitate reaching this goal. Among the many methods developed in the detection of biomarkers, aptamer-based biosensors (aptasensors) have shown great promise. Aptamers are promising diagnostic molecules with high sensitivity and selectivity, low-cost synthesis, easy modification, low toxicity, and high stability. Electrochemical aptasensors with high sensitivity and accuracy have attracted considerable attention in the field of biomarker detection. In this review, we will summarize recent advances in biomarker detection using electrochemical aptasensors. The principles of detection, sensitivity, selectivity, and other important factors in aptasensor performance are investigated. Finally, advantages and challenges of the developed aptasensors are discussed. [ABSTRACT FROM AUTHOR]

This work reports on further development of an optical biosensor for the in vitro detection of mycotoxins (in particular, aflatoxin B1) using a highly sensitive planar waveguide transducer in combination with a highly specific aptamer bioreceptor. This sensor is built on a SiO2–Si3N4–SiO2 optical planar waveguide (OPW) operating as a polarization interferometer (PI), which detects a phase shift between p- and s-components of polarized light propagating through the waveguide caused by the molecular adsorption. The refractive index sensitivity (RIS) of the recently upgraded PI experimental setup has been improved and reached values of around 9600 rad per refractive index unity (RIU), the highest RIS values reported, which enables the detection of low molecular weight analytes such as mycotoxins in very low concentrations. The biosensing tests yielded remarkable results for the detection of aflatoxin B1 in a wide range of concentrations from 1 pg/mL to 1 μg/mL in direct assay with specific DNA-based aptamers. [ABSTRACT FROM AUTHOR]

Abstract: Electrochemical aptasensors appear as promising tools in food analysis, able to provide sensitive, fast and cost‐effective analysis, with the added advantage of portability. Carbon nanomaterials and in particular carbon nanotubes and graphene are among the nanomaterials most often used to build electrochemical aptasensors due to their good electrical conductivity, large surface area and multiple functionalisation possibilities. This review aims to give an overview of the types of carbon nanomaterials and their composites which have been used to enhance the performance of electrochemical aptasensors. Examples are detailed for the biosensors which were tested with real food samples. In these aptasensors, carbon nanomaterials have played different roles, from facilitating the immobilization of high amounts of aptamer and enhancing the electroactive area of the sensors to roles as nanocarrier for signaling probes in amplification schemes or even as electroactive probes generating the output signal. The survey of recent literature shows a positive evolution towards increased aptasensor testing with food samples. However, many challenges remain related to the better characterization of nanomaterials used, clarifying the roles of specific components in multi‐component nanocomposites and widening the types of food matrices and analytes tested with the aptasensors. Although we are still far from knowing when these novel tools will replace classic analytical methods in food analysis, carbon nanomaterials will certainly continue to play an important role in the design of future electrochemical aptasensors for food analysis. [ABSTRACT FROM AUTHOR]

Recent years have witnessed an overwhelming integration of nanomaterials in the fabrication of biosensors. Nanomaterials have been incorporated with the objective to achieve better analytical figures of merit in terms of limit of detection, linear range, assays stability, low production cost, etc. Nanomaterials can act as immobilization support, signal amplifier, mediator and artificial enzyme label in the construction of aptasensors. We aim in this work to review the recent progress in mycotoxin analysis. This review emphasizes on the function of the different nanomaterials in aptasensors architecture. We subsequently relate their features to the analytical performance of the given aptasensor towards mycotoxins monitoring. In the same context, a critically analysis and level of success for each nano-aptasensing design will be discussed. Finally, current challenges in nano-aptasensing design for mycotoxin analysis will be highlighted. [ABSTRACT FROM AUTHOR]

Various methodologies have been reported in the literature for the qualitative and quantitative monitoring of mycotoxins in food and feed samples. Based on their enhanced specificity, selectivity and versatility, bio-affinity assays have inspired many researchers to develop sensors by exploring bio-recognition phenomena. However, a significant problem in the fabrication of these devices is that most of the biomolecules do not generate an easily measurable signal upon binding to the target analytes, and signal-generating labels are required to perform the measurements. In this context, aptamers have been emerged as a potential and attractive bio-recognition element to design label-free aptasensors for various target analytes. Contrary to other bioreceptor-based approaches, the aptamer-based assays rely on antigen binding-induced conformational changes or oligomerization states rather than binding-assisted changes in adsorbed mass or charge. This review will focus on current designs in label-free conformational switchable design strategies, with a particular focus on applications in the detection of mycotoxins. [ABSTRACT FROM AUTHOR]

Development of highly sensitive analytical procedures for food contaminants is one of the critical points in addressing new challenges related to food safety worldwide. Electrochemical aptamer-based sensors have been intensively investigated as potential analytical tools providing the desired portability, fast response, sensitivity and specificity in addition to lower cost and simplicity versus classical methods. The paper summarizes the aptasensors reported in the literature in the last 3 years, that have been used in applications related to food safety. New trends pertaining to increasing the sensitivity of detection by using nanomaterials and engineering of new aptamers are briefly discussed. With the recent development of new aptamers and following the lead of aptasensors devoted to biomedical field, the next years will witness an avalanche of new exciting electrochemical aptasensors for food safety. [ABSTRACT FROM AUTHOR]

The contamination of food and feed by mycotoxins has become an increasingly serious problem. Mycotoxins represent a major risk to human and animal health, as well as economics. Herein, we focus on Ochratoxin A (OTA), which is one of the most common mycotoxins contaminating feed and foodstuffs. OTA is a secondary metabolite produced by various Aspergillus and Penicillium strains. Upon ingestion, OTA has a number of acute and chronic toxic effects. It is nephrotoxic, teratogenic, immunosuppressive, and carcinogenic (group 2B). As a consequence, some regulatory limits have been introduced on the levels of OTA in several commodities. The toxic nature of OTA demands highly sensitive and selective monitoring techniques to protect human and animal health. As alternative to traditional analytical techniques, biochemical methods for OTA analysis have attained great interest in the last few decades. They are mainly based on the integration of antibodies or aptamers as biorecognition elements in sensing platforms. However, aptamers have gained more attention in affinity-based assays because of their high affinity, specificity, stability, and their easy chemical synthesis. In this brief review, we present an overview of aptamer-based assays and their applications in OTA purification and detection, appeared in the literature in the last five years. [ABSTRACT FROM AUTHOR]

Patulin contamination in fruits, vegetables, and their products is considered a serious health risk factor for food safety and human health. Thus, a rapid, simple detection method for patulin is becoming important, which could provide a tool for routine screening and food surveys. The objective of this study was to develop a sensitive aptamer-based lateral flow assay (FLA) using Streptavidin functionalized gold nanoparticles for sensitive patulin detection. An excellent dynamic range for patulin detection was obtained (2.7~139.8 ng/mL in the buffer and 7.07~359.5 ng/mL in the sample) with no affinity for other mycotoxins such as zearalenone (ZEN), ochratoxin A (OTA), aflatoxin B1 (AFB1), citrinin or tenuazonic acid (TEA). The limit of detection was 0.19 ng/mL in the buffer and 0.36 ng/mL in the real sample. The recoveries were 83.3% to 107.1%, with a satisfactory RSD value from 6.5% to 7.5%. Hence the established LFA could be used as a rapid, simple, on-site screening tool for PAT determination in apple juice. [ABSTRACT FROM AUTHOR]

The lateral flow assay (LFA) is an extensively used paper-based platform for the rapid and on-site detection of different analytes. The method is user-friendly with no need for sophisticated operation and only includes adding sample. Generally, antibodies are employed as the biorecognition elements in the LFA. However, antibodies possess several disadvantages including poor stability, high batch-to-batch variation, long development time, high price and need for ethical approval and cold chain. Because of these limitations, aptamers screened by an in vitro process can be a good alternative to antibodies as biorecognition molecules in the LFA. In recent years, aptamer-based LFAs have been investigated for the detection of different analytes in point-of-care diagnostics. In this review, we summarize the applications of aptamer technology in LFAs in clinical diagnostic rapid tests for the detection of biomarkers, microbial analytes, hormones and antibiotics. Performance, advantages and drawbacks of the developed assays are also discussed. [ABSTRACT FROM AUTHOR]

The combination between the specific recognition abilities of aptamers and the catalytic power of enzymes represents a powerful tool in bioanalysis to achieve the highly selective and sensitive detection of various molecules. Enzymes attached to biomolecules and nanomaterials facilitate the highly amplified detection of electroactive compounds. Meanwhile DNAzymes, small DNA strands showing enzyme-like activity are increasingly used as replacements of natural enzymes in electrochemical aptasensors. Moreover, nucleic acid catalysts have been integrated together with aptamer domains in aptazymes, where the aptamer-ligand interaction acts as a switch for the enzyme-like activity. The review discusses enzyme and DNAzyme based amplification, as the preferred approaches in electrochemical aptasensors. DNA and nanomaterial based signal enhancement is briefly mentioned, while a larger discussion is dedicated to aptasensors for the measurement of enzymatic activity and for the development of "universal" detection tools based on the aptamer-enzyme combination. This includes assays combining aptamers with glucometers for the detection of various molecules and promising "aptameric enzyme subunits"-yet to be integrated with electrochemical detection. The outlooks of the electrochemical biosensors coupling aptamers and enzymes are presented. [ABSTRACT FROM AUTHOR]

In this work, we describe the use of a new truncated aptamer for the determination of ofloxacin (OFL), being a principal quinolone commonly used in both human and animal healthcare. Since the affinity of a 72-mer ssDNA sequence has been previously described without further investigations, this paper demonstrates the first computational prediction of the binding motif between this aptamer and OFL through in silico molecular docking studies. Besides, we suggest the application of the characterized recognition mechanism in a simple FRET (Förster Resonance Energy Transfer) pattern for the rapid aptasensing of the quinolone of interest. Accordingly, our approach harnesses the fluorescence quenching of the fluorescein-tagged aptamer (FAM-APT) induced by its partial hybridization to a tetramethyl rhodamine-labelled complementary ssDNA (TAMRA-cDNA). In such a structure, dye labels brought into close proximity act as a FRET pair. Upon ofloxacin addition, an affinity competition occurs to form a more stable FAM-APT/OFL complex, thus unquenching the FAM-APT signal. Interestingly, the recovered fluorescence intensity was found to correlate well with the antibiotic's concentrations in the range of 0.2–200 μM in HEPES buffer, with a linear response that ranged between 0.2 and 20 μM. The rapid apta-assay achieved limits of detection and quantification of 0.12 and 0.40 μM, respectively. The truncated aptamer has also shown an improved specificity toward OFL than other quinolones, compared to the original full-length aptamer described in previous works. Finally, the practical application of the developed apta-assay was successfully confirmed to detect OFL quinolone in spiked milk samples, with satisfactory recoveries ranging between 97.4% and 111.4%. [ABSTRACT FROM AUTHOR]

This work focuses on the development of the novel label-free optical apta-sensors for detection of mycotoxins. A highly sensitive analytical method of total internal reflection ellipsometry (TIRE) combined with Localized Surface Plasmon Resonance (LSPR) phenomenon in nano-structured gold films was exploited here for the first time for detection of aflatoxin B1 and M1 in direct assay with specific aptamers immobilized on the surface of gold. The achieved detection of low molecular weight molecules, such as aflatoxin B1 and M1, in a wide range of concentrations from 100 ng/mL down to 0.01 ng/mL is remarkable for the LSPR method. The study of binding kinetics of aflatoxin molecules to their respective aptamers using dynamic TIRE measurements yielded the values of affinity constants in the range of 10−8–10−7 mol, which is characteristic for highly specific aptamer/target interactions similar to that for monoclonal antibodies. The effect of aptamers’ DNA chain length on their binding characteristics was analyzed. [ABSTRACT FROM AUTHOR]

This work demonstrates the development of electrochemical aptasensor using ochratoxin A (OTA) aptamers. Different aptamer coupling strategies were tested using polythiophene-carboxylic acid (PT3C) and polypyrrole-3-carboxylic acid (PP3C). The best sensitivity was recorded by polythiophene-3-carboxylic acid (PT3C) on screen-printed carbon electrode (SPCE) to attain the direct detection of OTA. The quantification of OTA was achieved by using electrochemical impedance spectroscopy. A good dynamic range 0.125–2.5 ng ml −1 was obtained for OTA with limit of detection (LOD) 0.125 ng ml −1 and Limit of quantification (LOQ) 0.3 ng ml −1 respectively. The good reproducibility was recorded with RSD% of 3.68. The obtained straight line equation was y = 0.4061 × + 1.03, r = 0.99. For real sample applications, the developed aptasensors were demonstrated in coffee samples. The aptasensor displayed good recovery values in the range 88–89%, thus exhibited the effectiveness of proposed aptasensor for such complex matrices. [ABSTRACT FROM AUTHOR]

Abstract: This work presents an aptasensor for Ochratoxin A (OTA) using unmodified gold nanoparticles (AuNPs) indicator. The assay method is based on the conformation change of OTA''s aptamer in phosphate buffered saline (PBS) containing Mg2+ and OTA, and the phenomenon of salt-induced AuNPs aggregation. A single measurement took only five minutes. Circular dichroism spectroscopic experiments revealed for the first time that upon the addition of OTA, the conformation of OTA''s aptamer in PBS buffer changed from random coil structure to compact rigid antiparallel G-quadruplex structure. This compact rigid G-quadruplex structure could not protect AuNPs against salt-induced aggregation, and thus the color change from red to blue could be observed by the naked eye. The linear range of the colorimetric aptasensor covered a large variation of OTA concentration from 20 to 625nM and the detection limit of 20nM (3σ) was obtained. [Copyright &y& Elsevier]

The current work is a continuation of our research targeted the development of novel optical sensing technologies for detection of mycotoxins. The method of (TIRE) was developed in the last decade as a combination of spectroscopic ellisometry and SPR and was proved to be a highly sensitive analytical tool in bio-sensing particularly attractive for detection of low molecular weight analytes, such as mycotoxins. The use of aptamers as highly specific artificial molecular receptors to ochratoxin A (OTA) in conjunction with the method Total Internal Reflection Ellipsometry (TIRE) is reported here for the first time. Our results showed a possibility of label-free optical detection of OTA down to 0.01 ppb in concentration in direct assay with specific aptamer. The kinetics of aptamer/OTA binding was studied with dynamic TIRE spectral measurements and allowed evaluating the affinity constant K D  = 1.8 10 −8  Mol which is characteristic for highly specific aptamer/OTA binding. [ABSTRACT FROM AUTHOR]

In this work, we developed an electrochemical aptasensor by using methylene blue (MB) redox probe labeled aptamer as a signaling fragment and functional graphene oxide (FGO) as the signal-enlarging platform. The role of functionalized graphene oxide was not mere to serve as a covalent immobilization support for aptamer sequences, but its catalytic signal amplification behavior towards methylene blue was demonstrated for the first time in the present work. The functionalized graphene oxide was cast on screen-printed carbon electrodes (SPCE), and then the MB-tagged aptamer was covalently immobilized on SPCE by using hexamethylenediamine (HMDA) as a spacer via carbodiimide amide-bonding chemistry. Aflatoxin B1 (AFB1) analyte molecule detection was accomplished by the aptamer conjugated redox probe, which undergoes/involves a conformational change in the complex structure of aptamer consequent to AFB1 binding. The proposed assay permitted to detect AFB1 in the linear range of 0.05–6.0 ng mL −1 with a very low limit of detection (LOD) (0.05 ng mL −1 ). The present principle aptasensor was tested to screen the alcoholic beverage samples for AFB1 detection and good recovery values were obtained. [ABSTRACT FROM AUTHOR]

The discovery of in-vitro systematic evolution of ligands by exponential enrichment (SELEX) process has considerably broaden the utility of aptamer as bio-recognition element, providing the high binding affinity and specificity against the target analytes. Recent research has focused on the development of structure switching signaling aptamer assay, transducing the aptamer- target recognition event into an easily detectable signal. In this paper, we demonstrate the development of structure switching aptamer assay for determination of aflatoxin M1 (AFM1) employing the quenching-dequenching mechanism. Hybridization of fluorescein labelled anti-AFM1 aptamer (F-aptamer) with TAMRA labelled complementary sequences (Q-aptamer) brings the fluorophore and the quencher into close proximity, which results in maximum fluorescence quenching. On addition of AFM1, the target induced conformational formation of antiparallel G-quadruplex aptamer-AFM1 complex results in fluorescence recovery. Under optimized experimental conditions, the developed method showed the good linearity with limit of detection (LOD) at 5.0ngkg-1 for AFM1. The specificity of the sensing platform was carefully investigated against aflatoxin B1 (AFB1) and ochratoxin A (OTA). The developed assay platform showed the high specificity towards AFM1. The practical application of the developed aptamer assay was verified for detection of AFM1 in spiked milk samples. Good recoveries were obtained in the range from 94.40% to 95.28% (n=3) from AFM1 spiked milk sample. Graphical abstractfx1Highlights· Structure switching aptamer assay for detection of aflatoxin M1 (AFM1). · Successfully investigated for detection of AFM1 in spiked milk samples. · Analytical performance of aptamer assay proves its applicability in milk analysis. · No complex milk sample preparation procedure. · Obtained recoveries were in the range of 94.40-95.28% (n=3). [ABSTRACT FROM AUTHOR]

This work demonstrates the development of a folding based electrochemical aptasensor using methylene blue (MB) tagged anti-Ochratoxin A (OTA) aptamers. Different aptamer coupling strategies were tested using Hexamethylenediamine, polyethylene glycol, simple adsorption and diazonium coupling mechanism. The best sensitivity was recorded by oxidation of amines using hexamethylenediamine (HDMA) on screen printed carbon electrode (SPCE). To achieve the direct detection of OTA, aptamer conjugated redox probe was used and detection was demonstrated based on the conformational changes in aptamer structure upon OTA sensing. Signaling in this class of sensors arises from changes in electron transfer efficiency upon target-induced changes in the conformation/flexibility of the aptamer probe. These changes can be readily recorded electrochemically. The developed aptasensor is unique in its own mechanism as redox probe tagged aptamer coupling such as MB has never been tried to immobilize using long carbon chain spacers as, addition of spacers would provide more sensitive detection methods. A good dynamic range 0.01–5 ng/ml was obtained for OTA with Limit of detection (LOD) 0.01 ng/ml and Limit of quantification (LOQ) of 0.03 ng/ml respectively. The good reproducibility was recorded with RSD% of 3.75. The obtained straight line equation was y =0.4035 x +0.90311, r =0.9976. We believe that the sensor design guidelines outlined here represents a general strategy for developing new folding-based electrochemical aptasensors. The developed aptasensor was extended to screen cocoa samples for OTA contamination. The cocoa samples were extracted and purified using molecular imprinted polymer (MIP) columns. The aptasensor displayed good recovery values in the range 84–85% thus, exhibited the effectiveness of proposed aptasensor for such complex matrices. [ABSTRACT FROM AUTHOR]

Abstract: We report a new label-free colorimetric aptasensor based on DNAzyme–aptamer conjugate for rapid and high-throughput detection of Ochratoxin A (OTA, a possible human carcinogen, group 2B) in wine. Two oligonucleotides were designed for this detection. One is N1 for biorecognition, which includes two adjacent sequences: the OTA-specific aptamer sequence and the horseradish peroxidase (HRP)-mimicking DNAzyme sequence. The other is a blocking DNA (B2), which is partially complementary to a part of the OTA aptamer and partially complementary to a part of the DNAzyme. The existence of OTA reduces the hybridization between N1 and B2. Thus, the activity of the non-hybridized DNAzyme is linearly correlated with the concentration of OTA up to 30nM with a limit of detection of 4nM (3σ). Meanwhile, a double liquid–liquid extraction (LLE) method is accordingly developed to purify OTA from wine. Compared with the existing HPLC-FD or immunoassay methods, the proposed strategy presents the most appropriate balance between accuracy and facility, resulting in a considerable improvement of real-time quality control, and thereby, preventing chronic poisoning caused by OTA contained red wine. [Copyright &y& Elsevier]

Abstract: We report an aptasensor for biosensing of Ochratoxin A (OTA) using aptamer-DNAzyme hairpin as biorecognition element. The structure of this engineered nucleic acid includes the horseradish peroxidase (HRP)-mimicking DNAzyme and the OTA specific aptamer sequences. A blocking tail captures a part of these sequences in the stem region of the hairpin. In the presence of OTA, the hairpin is opened due to the formation of the aptamer–analyte complex. As a result, self-assembly of the active HRP-mimicking DNAzyme occurs. The activity of this DNAzyme is linearly correlated with OTA concentration up to 10nM, showing a limit of detection of 2.5nM. [Copyright &y& Elsevier]