Aptamer Probe Research Articles

Aptamer Probes Labeled with Lanthanide-Doped Carbon Nanodots Permit Dual-Modal Fluorescence and Mass Cytometric Imaging.

High‐dimensional imaging mass cytometry (IMC) enables simultaneous quantification of over 35 biomarkers on one tissue section. However, its limited resolution and ultralow acquisition speed remain major issues for general clinical application. Meanwhile, conventional immunofluorescence microscopy (IFM) allows sub‐micrometer resolution and rapid identification of the region of interest (ROI), but only operates with low multiplicity. Herein, a series of lanthanide‐doped blue‐, green‐, and red‐fluorescent carbon nanodots (namely, B‐Cdots(Ln1), G‐Cdots(Ln2), and R‐Cdots(Ln3)) as fluorescence and mass dual‐modal tags are developed. Coupled with aptamers, B‐Cdots(159Tb)‐A10‐3.2, G‐Cdots(165Ho)‐AS1411, and R‐Cdots(169Tm)‐SYL3C dual‐functional aptamer probes, which are then multiplexed with commercially available Maxpar metal‐tagged antibodies for analyzing clinical formalin‐fixed, paraffin‐embedded (FFPE) prostatic adenocarcinoma (PaC) tissue, are further synthesized. The rapid identification of ROI with IFM using fluorescence signals and subsequent multiplexed detection of in situ ROI with IMC using the same tissue section is demonstrated. Dual‐modal probes save up to 90% IMC blind scanning time for a standard 3.5 mm × 3.5 mm overall image. Meanwhile, the IFM provides refined details and topological spatial distributions for the functional proteins at optical resolution, which compensates for the low resolution of the IMC imaging.

A disposable gold foil paper-based aptasensor for detection of enteropathogenic Escherichia coli with SERS analysis and magnetic separation technology.

Rapid and sensitive detection of enteropathogenic Escherichia coli (EPEC) in fluids with complex background is an important task for safety quality control in the field of medicine, environment, and food. In this study, a gold foil paper-based aptasensor was developed for the detection of enteropathogenic EPEC O26:K60 with surface-enhanced Raman spectroscopy(SERS) and magnetic separation technology mediated by Fe3O4@Au composite. The gold foil paper was firstly modified with thiolated capture probe and SERS tag. Thethiolated aptamer probe for EPEC was immobilized onto a Fe3O4@Au composite. In the presence of EPEC, highly specific recognition between the aptamer probe and EPEC made the Fe3O4@Au composite partially dissociated from the gold foil paper. This led to a decreased Raman intensity response, which showed an obvious negative linear correlation with increasing concentration of EPEC over a wide concentration range from 10 to 107CFU/mL under an excitation wavelength of 633nm. The detection limit was about2.86CFU/mL in a buffer solution and a licorice extractum and the detection time was only 2.5h. Theresults demonstrate that the gold foil paper-based aptasensor can be an excellent biosensing platform that offers a reliable, rapid, and sensitive alternative for EPEC detection.

Anti-cancer adjuvant drug screening via epithelial-mesenchymal transition-related aptamer probe.

Epithelial-mesenchymal transition (EMT) is implicated in the pathological processes of cancer metastasis and drug resistance. Anti-cancer drugs may also potentially lead to EMT, resulting in their reduced therapeutic effect. Therefore, the combination of these anti-cancer drugs with anti-EMT agents has been promoted in clinic. Screening anti-EMT drugs and evaluation of EMT process are highly dependent on EMT biomarkers on cell membrane. At present, the detection of EMT biomarker is mainly by Western blot method, which is time-consuming and complicated. In this work, for effectively screening anti-EMT drugs by evaluation of the EMT process, a type of aptamer probe based on aggregation-induced emission (AIE) was designed. The aptamer SYL3C was employed to target the EMT biomarker EpCAM on cell membrane. Two fluorophores, FAM and tetraphenylethene (TPE, an AIE dye), were modified at the two ends of SYL3C, respectively. This aptamer probe (TPE-SYL3C-FAM) can monitor the EpCAM expression, which can be recovered by anti-EMT drugs. By observation of the change in TPE emission intensity, the anti-EMT effect of drugs can be evaluated. The FAM emission was used as internal reference to reduce environmental interferences. This probe can be potentially used to screen anti-EMT agents as anti-cancer adjuvant drugs with high throughput.

Introducing an SPRi-based titration assay using aptamers for the detection of Legionella pneumophila

Legionella are waterborne bacteria and the causative agents behind a severe respiratory disease known as Legionnaires Disease (LD). Standard ISO-certified methods to detect Legionella (plate count and qPCR) are time-consuming and tedious necessitating the development of novel detection strategies. This study introduces an SPRi-based titration assay that uses L. pneumophila (Lp) aptamers. Multiple parameters including buffer conditions, aptamer concentrations, complementary aptamer probe (cApt) concentrations and incubation times were investigated and evaluated for optimal signal response. For maximum hybridization efficiency on the sensor surface, high salt buffer (SSC) as well 500 nM of immobilized cApt resulted in a maximum signal response. Additionally, it was observed that of the three buffers (PBS, Fraquil, SSC) tested in this assay only SSC facilitated incremental binding of aptamers to Lp such that varying concentrations of Lp could be determined. We were able to detect varying concentrations of Lp down to a limit of 104 cells/ml without any sample processing or signal amplification steps, highlighting the potential of this system for SPRi-based Lp detection. The assay is also specific to Lp and shows negligible detection of Pseudomonas, a common inhabitant of water systems.

Label-free detection of Cu(II) in fish using a graphene field-effect transistor gated by structure-switching aptamer probes

Copper sulfate is a widely used agent to control insects, bacteria and algae for fishery. However, excess amount of copper ions in water accumulate in aquatic products through the ecological cycle system, highly threatening food safety and public health. Therefore, it is urgent to develop a rapid and efficient method for the determination of copper content in aquatic products. In this study, we developed a label-free biosensor for Cu(II) based on a graphene field-effect transistor gated by structure-switching aptamer probes (SSA-GFET) against Cu(II) we obtained before. The detection mechanism of the biosensor is attributed to the surface charge shift and the potential change of the gate electrode upon the specific binding of Cu(II). The SSA-GFET biosensor has a low detection limit of 10 nM and a linear range of 10 nM to 3 μM to Cu(II). In addition to the excellent selectivity to Cu(II), the biosensor also showes the advantage of high recovery rate for detection of Cu(II) in real fish samples. Because of the detection characteristics of label-free SSA-GFET, it has great advantages in the field of food safety and environmental detection.

Modular fluorescent nanoparticle DNA probes for detection of peptides and proteins

Fluorescently labeled antibody and aptamer probes are used in biological studies to characterize binding interactions, measure concentrations of analytes, and sort cells. Fluorescent nanoparticle labels offer an excellent alternative to standard fluorescent labeling strategies due to their enhanced brightness, stability and multivalency; however, challenges in functionalization and characterization have impeded their use. This work introduces a straightforward approach for preparation of fluorescent nanoparticle probes using commercially available reagents and common laboratory equipment. Fluorescent polystyrene nanoparticles, Thermo Fisher Scientific FluoSpheres, were used in these proof-of-principle studies. Particle passivation was achieved by covalent attachment of amine-PEG-azide to carboxylated particles, neutralizing the surface charge from − 43 to − 15 mV. A conjugation-annealing handle and DNA aptamer probe were attached to the azide-PEG nanoparticle surface either through reaction of pre-annealed handle and probe or through a stepwise reaction of the nanoparticles with the handle followed by aptamer annealing. Nanoparticles functionalized with DNA aptamers targeting histidine tags and VEGF protein had high affinity (EC50s ranging from 3 to 12 nM) and specificity, and were more stable than conventional labels. This protocol for preparation of nanoparticle probes relies solely on commercially available reagents and common equipment, breaking down the barriers to use nanoparticles in biological experiments.

Study on the binding mode of aptamer to ampicillin and its electrochemical response behavior in two different reaction media.

A study was carried out to investigate the binding mode of aptamer to ampicillin (AMP) and its electrochemical response behavior. The binding mode was confirmed using the molecular dynamics (MD) simulation method to obtain the corresponding binding dynamic change process. Following the confirmed binding mode, a qualitative elucidation was provided on the electrochemical response characteristics of a single-probe aptamer-based folding sensor. The results show that there exist two different binding modes in two different solution systems, Phys2 and H2O (0.1M NaCl). These two binding modes can respectively induce two different contraction changes, thereby driving the methylene blue (MB)-modified aptamer probe to show a "close-to-interface" convergence behavior with different degrees on the actual electrode surface, which validates two apparently different electrochemical response behavior characteristics of "signal-on" for the sensor. By contrast, H2O (0.1M NaCl) as the reaction medium is more conducive to the formation of a stable aptamer/AMP complex and the development of a high-sensitivity analytical method with a low detection limit of 0.033μM. The simulation results effectively support the experimental results, which is helpful in gaining a deeper understanding of the relationship between the signaling mechanism and practical analytical performance for aptamer-based folding sensors at the molecular level.

Dual-ratiometric electrochemical aptasensor enabled by programmable dynamic range: Application for threshold-based detection of aflatoxin B1

Aptamer-based sensor with high-specificity typically has a fixed linear range due to the host-guest binding. To expand the dynamic range for multiple scenarios, we report here a dual-ratiometric electrochemical aptasensor that integrates two aptamer profiles with varied affinity into a single sensing interface for aflatoxin B1 (AFB1) detection. Using functional aptamer as recognition element and generator of signals, we fabricate the dual-ratiometric aptasensor with anthraquinone loaded reduced graphene oxide (AQ-rGO), methylene blue labeled hairpin DNA (MB-DNA), and ferrocene labeled linear aptamer (Fc-apt), using MB and Fc probes to work in tandem while AQ as reference. The current of Fc (IFc) is used to detect the low-concentration analyte, and that of MB (IMB) varies when the concentration of AFB1 reaches a threshold. The threshold switch for IMB could be tuned by engineering the quantity ratio of Fc-apt and MB-DNA (a ratio of 1:1 is used here). The aptasensor can rapidly achieve the positive (+)/negative (-) detection of AFB1 with a threshold concentration of 10 pg mL-1, while the quantitative detection employs the ratio of current of AQ, MB, and Fc (i.e., IAQ/IMB and IAQ/IFc) as yardsticks, offering two linear ranges of 10-106 and 10-2-5 × 104 pg mL-1, respectively. The aptasensor is successfully used to monitor the amount of AFB1 in a 7-days mildew process of peanut. Such a protocol opens a new way for the design of sensors with the programmable linear range in the advanced biological and chemical sensing.

A highly specific aptamer probe targeting PD-L1 in tumor tissue sections: Mutation favors specificity

Although DNA aptamers can show comparable affinity to antibodies and have the advantage of having high batch-to-batch consistency, they often suffer from unsatisfied specificity for complex samples. The limited library size used for aptamer in vitro isolation (SELEX) has been recognized as one of the major reasons. Programmed cell death-ligand 1 (PD-L1) is both a key protein in cancer diagnostics and also immunotherapy. We report here a DNA aptamer that highly specifically binds PD-L1 expressed on the surface of various cancer cells and multiple types of tissue sections. The aptamers were selected from a DNA library containing a type II restriction endonuclease Alu I recognition site in the middle of the 40-nt random sequences, against recombinant PD-L1 rather than the whole cell or tissue section. The library enrichment was achieved by Alu I mediated-SELEX, named as REase-SELEX, in which Alu I cut off the non-binders at the recognition site and, more importantly, induced library mutations to substantially increase the library diversity. 8–60, a representative aptamer with high affinity (KD = 1.4 nM determined by SPR) successfully detected four types of cancer cells with PD-L1 expression levels from low to high by flow cytometry, normal human tonsil (gold standard for PD-L1 antibody evaluation), clinical non-small cell lung cancer (high PD-L1 expression level), and malignant melanoma (low PD-L1 expression level) tissue sections by fluorescence microscopy imaging, showing unprecedented high specificity. The results demonstrate that 8–60 is an advanced probe for PD-L1 cancer diagnostics and mutations in SELEX greatly favor aptamer specificity.

Proximity ligation assay mediated rolling circle amplification strategy for in situ amplified imaging of glycosylated PD-L1.

Glycosylated PD-L1 is a more reliable biomarker for immune checkpoint therapy and plays important roles in tumor immunity. Glycosylation of PD-L1 hinders antibody-based detection, which is partially responsible for the inconsistency between PD-L1 immunohistochemical results and therapeutic treatment response. Herein, we present a proximity ligation assay mediated rolling circle amplification (PLA-RCA) strategy for amplified imaging of glycosylated PD-L1 in situ. The strategy relies on a pair of DNA probes: an aptamer probe to specifically recognize cellular surface protein PD-L1 and a glycan conversion (GC) probe for metabolic glycan labeling. Upon proximity ligation of sequence binding to the two probes, the proximity ligation-triggered RCA occurs. The feasibility of the as-proposed strategy has been validated as it realized the visualization of PD-L1 glycosylation in different cancer cells and the monitoring of the variation of PD-L1 glycosylation during drug treatment. Thus, we envision the present work offers a useful alternative to track protein-specific glycosylation and potentially advances the investigation of the dynamic glycan state associated with the disease process.

Target-receptive structural switching of ssDNA as selective and sensitive biosensor for subsequent detection of toxic Pb2+ and organophosphorus pesticide

A structural switching in Single-stranded DNA (ssDNA) fluorescence biosensor for quick turn-on/off detection of Pb2+ ions and pesticide was reported. The design strategy of Hex-labelled ssDNA consists of two types of aptamer probe, G-rich base pair sequence forms G-quadruplex confirmation with Pb2+ ions. While other part of base pair sequence exhibits affinity to fold isocarbophos pesticide. MoS2 nanosheets were identified as quick quencher of Hex fluorescence intensity via Vander-Waals interaction and its significance was compared with other nanomaterials. This sensing mechanism proposes a specific affinity of GA-rich ssDNA with Pb2+ to form G-quadruplex via G-Pb2+-G sequences. Consequently, ssDNA relived from MoS2 nanosheets to restore the fluorescence intensity (turn-on). Subsequent addition of pesticide shows stronger affinity towards unfolded aptamer probe to form a random coil like structure. This causes Hex-labelled 5′ end closer to the G-quadruplex connected at the 3’ end of ssDNA resulting in a remarkable fluorescence quenching (turn-off) owing to PET process. Moreover, the sensing probe (Hex-labelled GA-rich ssDNA) was recycled by introducing acetylcholinesterase enzyme and thiocoline into the reaction mixture. The detection limits of Pb2+ and isocarbophos pesticide was estimated to be 0.6 nM and 0.018 μg/L respectively. Moreover, this study reveals a high sensitivity and selectivity towards target molecules in environmental samples.

Dual-Multivalent-Aptamer-Conjugated Nanoprobes for Superefficient Discerning of Single Circulating Tumor Cells in a Microfluidic Chip with Inductively Coupled Plasma Mass Spectrometry Detection.

The efficient recognition of circulating tumor cells (CTCs) with an aptamer probe confers numerous benefits; however, the stability and binding affinity of aptamers are significantly hampered in real biological sample matrices. Inspired by the efficient preying mechanism by multiplex tubing feet and endoskeletons of sea urchins, we engineered a superefficient biomimetic single-CTC recognition platform by conjugating dual-multivalent-aptamers (DMAs) Sgc8 and SYL3C onto AuNPs to form a sea urchin-like nanoprobe (sea urchin-DMA-AuNPs). Aptamers Sgc8 and SYL3C selectively bind with the biomarker proteins PTK7 and EpCAM expressed on the surface of CTCs. CTCs were captured with 100% efficiency, followed by sorting on a specially designed multifunctional microfluidic configuration, integrating a single-CTC separation unit and a hydrodynamic filtrating purification unit. After sorting, background-free analysis of biomarker proteins in single CTCs was undertaken with inductively coupled plasma mass spectrometry by measuring the amount of 197Au isotope in sea urchin-DMA-AuNPs. With respect to a single-aptamer nanoprobe/-interface, the dual-aptamer nanoprobe improves the binding efficiency by more than 200% (Kd < 0.35 nM). The microchip facilitates the recognition of single CTCs with a sorting separation rate of 93.6% at a flow rate of 60 μL min-1, and it exhibits 73.8 ± 5.0% measurement efficiency for single CTCs. The present strategy ensures the manipulation and detection of a single CTC in 100 μL of whole blood within 1 h.

Ratiometric G-Quadruplex Assay for Robust Lead Detection in Food Samples.

Lead (Pb2+) pollution is a serious food safety issue, rapid detection of Pb2+ residual in food is vital to guarantee food quality and safety. Here we proposed ratiometric aptamer probes, allowing robust Pb2+ supervision in food samples. Pb2+ specific aptamer can bolster a transition of G-quadruplex structural response to Pb2+; this process can be monitored by N-methyl mesoporphyrin IX (NMM), which is highly specific to G-quadruplex. Particularly, the utilization of G-quadruplex specific dye and terminal-labeled fluorophore allowed to endue ratiometric signal outputs towards Pb2+, dramatically increase the robustness for lead detection. The ratiometric G-quadruplex assay allowed a facile and one-pot Pb2+ detection at room temperature using a single-stranded DNA aptamer. We demonstrated its feasibility for detecting lead pollution in fresh eggs and tap water samples. The ratiometric G-quadruplex design is expected to be used for on-site Pb2+ testing associated with food safety.

Aptamer-Sensitized Nanoribbon Biosensor for Ovarian Cancer Marker Detection in Plasma

The detection of CA 125 protein in buffer solution with a silicon-on-insulator (SOI)-based nanoribbon (NR) biosensor was experimentally demonstrated. In the biosensor, sensor chips, bearing an array of 12 nanoribbons (NRs) with n-type conductance, were employed. In the course of the analysis with the NR biosensor, the target protein was biospecifically captured onto the surface of the NRs, which was sensitized with covalently immobilized aptamers against CA 125. Atomic force microscopy (AFM) and mass spectrometry (MS) were employed in order to confirm the formation of the probe–target complexes on the NR surface. Via AFM and MS, the formation of aptamer–antigen complexes on the surface of SOI substrates with covalently immobilized aptamers against CA 125 was revealed, thus confirming the efficient immobilization of the aptamers onto the SOI surface. The biosensor signal, resulting from the biospecific interaction between CA 125 and the NR-immobilized aptamer probes, was shown to increase with an increase in the target protein concentration. The minimum detectable CA 125 concentration was as low as 1.5 × 10−17 M. Moreover, with the biosensor proposed herein, the detection of CA 125 in the plasma of ovarian cancer patients was demonstrated.

Aptasensing luteinizing hormone to determine gynecological endocrine complications on graphene oxide layered sensor.

Luteinizing hormone (LH)/lutropin is an interstitial cell-stimulating hormone playing a predominant role in the reproductive system, and highly correlated with the infertility treatment in both men and women. This research was concentrated to quantify LH level by using interdigitated electrode sensor. To improve the electric current flow, sensing electrode was modified with graphene oxide (GO) and the aptamer probe was attached on GO through biotin-streptavidin linker. Current responses were measured with aptamer-LH interaction at the target concentrations between 7.5nM and 1μM and the detection limit of LH was calculated as 60nM with the determination coefficient (R2 ) value, 0.9229 [y = 1.296x - 2.8435] on a linear range from 30nM to 1μM. Further, biofouling effect on sensing electrode surface was analyzed with complementary aptamer sequence, control proteins (albumin and globulin). The above GO-aptamer-modified interdigitated electrode sensor helps to quantify LH level and diagnose gynecological endocrinology-related complications.

A photo-electrochemical aptasensor for the determination of severe acute respiratory syndrome coronavirus 2 receptor-binding domain by using graphitic carbon nitride-cadmium sulfide quantum dots nanocomposite

Herein, a photoelectrochemical aptasensor for the quantitive measurement of the severe acute respiratory syndrome coronavirus-2 receptor-binding domain (Sars-Cov-2 RBD) has been reported for the first time. For this purpose, first, graphitic carbon nitride and (gC3N4) and cadmium sulfide (CdS) quantum dots were fabricated and characterized. After that, gC3N4 and CdS were mixed well. The fabricated nanomaterials were characterized by scanning transmission electron microscopy. Then, the CdS QDs-gC3N4 nanocomposite was added to the solution containing chitosan as an amine-rich polymer to generate a Chitosan/CdS-gC3N4 nanocomposite. Subsequently, the surface of the ITO electrode was modified with Chitosan/CdS-gC3N4. After that, the amine-terminal aptamer probes were immobilized on the surface of the Chitosan/CdS QDs-gC3N4/ITO electrode by using glutaraldehyde as an amine-amine crosslinker. The electrochemical performances of the electrodes were studied using cyclic voltammetry (CV), electrochemical Impedance Spectroscopy (EIS), and photo-electrochemistry (PEC). The surface coverage of the immobilized aptamer probe was founded to be 26.2 pmol.cm−2. The obtained results demonstrated that the proposed photo-electrochemical aptasensor can be used for the measurement of Sars-Cov-2 RBD within 0.5−32.0 nM. The limit of detection (LOD) was obtained to be 0.12 nM (at 3σ/slope). The affinity of the Aptamer/Chitosan/CdS QDs-gC3N4/ITO was also founded to be 3.4 nM by using Langmuir-typical adsorption systems. The proposed photo-electrochemical aptasensor was applied for the measurement of the spiked Sars-Cov-2 RBD in human saliva samples at two concentrations. The effect of the interfering biomaterials such as human immunoglobulin G human immunoglobulin A, human immunoglobulin M, and human serum albumin was also studied.

Spectrally filtered photodiode pairs for on-chip ratiometric aptasensing of cytokine dynamics

Miniaturized, cost-effective cytokine sensing devices are essential for health monitoring and critical care of patients with acute immune responses. Here we report an on-chip ratiometric aptasensing device to monitor the dynamics of an essential immune response biomarker, interferon-gamma. Our device is formed by pairs of spectrally filtered silicon photodiodes that are surface functionalized with aptamer probes. These photodiode pairs are able to rapidly detect two-color fluorescence changes transduced from aptamer-cytokine binding events within c.a. 6 min at picomolar levels, which are sufficient to capture the cytokine transients in point-of-care settings. Furthermore, our device can be reset to baseline by simple washing steps, yielding consistent readout in consecutive runs. Combined with their chip-scale construct and real-time operation, our cytokine sensors may have possible use in point-of-care and therapeutic screening applications.

Aptamer-decorated porous microneedles arrays for extraction and detection of skin interstitial fluid biomarkers

The detection of biomarkers in body fluids plays a great role in the diagnosis, treatment, and prognosis of diseases. Here, we present novel aptamer-decorated porous microneedles (MNs) arrays to realize the extraction and detection of biomarkers in skin interstitial fluid (ISF) in situ. The porous MNs arrays are fabricated by replicating the negative molds comprising glass microspheres with a UV-curable ethoxylated trimethylolpropane triacrylate (ETPTA). As the MNs arrays combine the superiorities of porous structure and aptamers, their specific surface area increased significantly to 6.694 m2/g, thus vast of stable aptamer probes with a concentration of 0.9459 μM could be immobilized. In addition, the MNs arrays could extract skin ISF into their porous structure on the basis of the capillarity principle, and subsequently capture and detect skin ISF biomarkers without sample post-process. Taking advantage of these features, we further demonstrated a highly sensitive and rapid detection of ISF endotoxin in the concentration ranges of 0.0342 EU/mL to 8.2082 EU/mL from rats model injected with endotoxin via tail vein by using such aptamer-decorated porous MNs arrays, with the limit of detection (LOD) of 0.0064 EU/mL. These results indicated that the aptamer-decorated porous MNs arrays possess great potential for non-invasive extraction and detection of biomarkers in clinical applications.

The role of band structure in Co- and Fe-co-doped Ba0.5Sr0.5Zr0.1Y0.1O3-δ perovskite semiconductor to design an electrochemical aptasensing platform: application in label-free detection of ochratoxin A using voltammetry.

Nanocomposites can offer a platform to conjugate biorecognition features of aptamer with unique size-dependent properties of a given material, which can autoprobe the binding event based on their electroactive characteristics. Herein, we design electroactive switchable aptamer probes based on co-doped single-phase semiconducting materials employing the cyclic voltammetry method to record the current signal at each step of electrochemical characterization. To do so, we utilized a facile hydrothermal method assisted by co-precipitation method such as Co-Fe-co-doped Ba0.5Sr0.5Zr0.1Y0.1O3-δ (CF-BSZY) and tuned the alignment of the energy band structure of the material to amplify the output of theelectrochemical signal. Atvarious steps, changes occurred in the electrochemical properties at the surface of CF-BSZY. The binding of the ssDNA with prepared materials enhances the current signal by the interaction with the target (ochratoxin A (OTA)) depressing the current signal and facilitating the construction of a novel design of electrochemical aptasensor. As a proof of concept, an electrochemical aptasensor for the detection of ochratoxin A (OTA) in rice samples has been developed. The electrochemical aptasensor provides a limit of detection (LOD) of 0.00012μM (0.12nM), with a linear range from 0.000247 to 0.74μM and sound OTA recovery in real samples. The developed aptasensor is simply designed andis free of oligonucleotide labeling or decorative nanoparticle modifications. The proposed mechanism is generic in principle with the potential to translate any type of aptamer and target binding event into a detectable signal; hence, it can be largely applied to various bioreceptor recognition phenomena for subsequent applications.

Electrochemical aptasensor based on Mo2C/Mo2N and gold nanoparticles for determination of chlorpyrifos.

Two-dimensional Mo2C/Mo2N composites were synthesized by high temperature ball milling and used as support materials for fabricating achlorpyrifos (CPF) aptasensor. Gold nanoparticles (Au NPs) were electrodeposited on the surface of aMo2C/Mo2N-modified electrode to connect with the ferrocene (Fc) probe via Au-S bonds. The Fc probe can hybridize with the aptamer probe to form a double-stranded structure. The addition of CPF made the double strands melt and the Fc probe approached the surface of the electrode, thereby resulting in amplification of the electrochemical response. The current response of the aptasensor for detecting CPF in solutions linearly varied from 0 to 400ngmL-1 (with a maximum at 0.98V vs. Ag/AgCl). The Au NPs/Mo2C/Mo2N composites exhibited satisfactory electrochemical behavior due to their excellent electrical conductivity and large surface area. This ultrasensitive aptasensor showed a low limit of detection of 0.036ngmL-1. It was applied to determine CPF in real samples with acceptable recoveries from 94.7 to 116.7%, and the relative standard deviation was from 2.57 to 7.08%.Graphical abstract Schematic diagram of the manufacturing process of the aptasensor. Electrochemical aptasensor based on Mo2C/Mo2N/Au NP composites show excellent performance in detecting CPF.