Electrochemical Aptasensor Research Articles

Dually-amplified electrochemical aptasensor based on the self-linking AuPt nanoflowers for ultrasensitive determination of hemagglutinin

BackgroundInfluenza virus can spread from person to person and cause epidemics. Therefore, rapid and sensitive diagnosis of virus is essential in controlling influenza outbreaks. Conventional virus diagnostic techniques are time-consuming, labor-intensive and requires large instruments. In this work, a sandwich electrochemical assay by a pair of aptamers was developed for ultrasensitive determination of hemagglutinin (HA) protein, which is one of the two surface glycoproteins of influenza A (H1N1) virus, using dual signal amplification techniques. ResultsHA was captured and magnetically separated by Fe3O4@SiO2–NH2@Au attached to aptamer 1 (Apt1), creating a sandwich structure with AuPt nanoflowers (AuPtNFs) connected to aptamer 2 (Apt2). Herein, AuPtNFs could catalyze H2O2/hydroquinone (HQ) to generate 1,4-benzoquinone (BQ), and achieved amplification of electrochemical signal detection through differential pulse voltammetry (DPV). The constructed aptasensor expressed a wide linear range (10 pg/mL-100 ng/mL) with limit of detection (LOD) of 2.4 pg/mL. Moreover, a novel strategy for dual signal amplification was developed to further enhance sensitivity. The innovative electrochemical aptasensor could achieve secondary amplification of the detection signal with LOD of 0.3 pg/mL and linear concentration range from 0.5 pg/mL to 100 ng/mL. The secondary amplification could be achieved only through the self-linking process, which allowed for the retention of numerous AuPtNFs by simple complementary base pairing to connect more AuPtNFs onto the above-mentioned sandwich structure. SignificanceOverall, the constructed aptasensor exhibited favorable sensitivity and accuracy, indicating the potential expanded application for the clinical detection of numerous viruses.

Design of a triple-functional polyA-probe: Toward a facile electrochemical aptasensor for vanillin detection

This study successfully developed an electrochemical aptasensor based on a triple-functional biotin-modified poly-adenine (polyA) probe (Bio-TPP) for one-step ultrasensitive detection of vanillin (VAN). The Bio-TPP was self-assembled onto a gold electrode (AuE) via its polyA sequence, followed by conjugation with PtPd@Ni-Co layered double hydroxides labeled with streptavidin and thionine (SA/Thi/PtPd@Ni-Co LDH), which generated electrochemical signals. Importantly, in the presence of VAN, the specific substrate sequence of a Mg²⁺-dependent DNAzyme in the Bio-TPP hybridized with the output DNAzyme strand generated by the specific recognition between the aptamer (Apt) and VAN. And, with the help of Mg²⁺, the DNAzyme amplification reaction was triggered. This led to a significant reduction in the electrochemical signals, enabling highly sensitive detection of VAN. Benefiting from the precise and flexible design of the Bio-TPP, this aptasensor offered a new detection method, exhibiting excellent performance with a wide linear range of 1×10−6-10 μM and a detection limit of 0.30 pM. Additionally, this assay possessed good selectivity, reproducibility, and stability, which held great potential in bioanalysis.

A double-conducting polymer nanowire-based electrochemical aptasensor for highly sensitive and low fouling detection of cortisol in saliva.

Acortisol biosensor was developedbased on double-conducting polymer nanowires, which exhibits excellent conductivity, resistance to biological contamination, and outstanding sensing performance. The biosensor employs dual-mode electrochemical techniques, namely, differential pulse voltammetry (DPV) and chronoamperometry (CA), for the sensitive and low fouling detection of the glucocorticoid hormonecortisol. Experimental results demonstrated that the linear detection range of the biosensor in DPV mode was 1.0 × 10-14-1.0 × 10-8M, with a detection limit of 0.131 × 10-14M. In CA mode, the biosensor exhibited a detection range of 1.0 × 10-13-1.0 × 10-7M and a detection limit of 0.313 × 10-13M. The biosensor was successfully utilized for the rapid detection of cortisol in human saliva. The combination of a high-specificity cortisol aptamer and functionalized double-conducting polymer nanowires ensured the exceptional specificity and sensitivity of the biosensor in detecting real biological samples.

Glutathione engineered gold nanoparticles-based electrochemical aptasensor for determination of arsenic ions in water, food, and soil samples

Arsenic, As(III) is one of the highly toxic metalloid ions present in the earth's crust. Its availability especially in groundwater, soil, and plant products can impose significant environmental and health risks. In order to examine aforementioned matrices for the presence of As(III), we have fabricated a sensitive electrochemical bionanoprobe using As(III) specific aptamer modified glutathione (GSH)-capped gold nanoparticles. The developed probe demonstrated remarkable sensing performance for a wide concentration range of As(III) (e.g., 0.1–1000 mg/L) with extremely high sensitivity of 0.0875 µA/(mg/L.mm2). Moreover, we have achieved an exceptionally low limit of detection (0.13 µg/L) for As(III) using the developed bionanoprobe. Furthermore, it displayed excellent performance in monitoring As(III) in groundwater, soil, and rice samples.

A Highly Sensitive Electrochemical Aptasensor for Kanamycin: Leveraging RecJf Exonuclease-Assisted Target Recycling and Hybridization Chain Reaction Signal Amplification

A Highly Sensitive Electrochemical Aptasensor for Kanamycin: Leveraging RecJf Exonuclease-Assisted Target Recycling and Hybridization Chain Reaction Signal Amplification

Portable label-free electrochemical aptasensor for sensitive detection of paraquat herbicide mediated by oxygen reduction reaction

Paraquat (PQ) is a highly toxic herbicide that has been prohibited in almost 70 countries, but remains in use worldwide. Thus, routine on-site PQ monitoring is a key mechanism to ensure safety and efficiently enforce regulations. Herein, a label-free portable electrochemical aptasensor for the detection of PQ was developed by utilizing aptamer designed to specifically recognize PQ. The aptasensor employs square-wave voltammetry (SWV) to quantify PQ binding on the aptamer-functionalized electrode surface by tracking the downstream oxygen reduction reaction. It provided a detection range spanning from 0.01 to 100.0 μg mL−1 PQ with a limit of detection (LOD) of 8.9 ng mL−1. Validation against spiked tap water, pomegranate juice, and orange juice revealed recovery rate performances of 75 %–130 %. The aptasensor demonstrates promising feasibility for PQ detection in real-world applications, offering remarkable portability and operational simplicity. Notably, it can operate without supplementary redox agents, requiring only sample incubation and subsequent washing steps.

Design and preparation of a sensitive electrochemical aptasensor based on hierarchical porous UiO-66 for detecting ampicillin

Hierarchical porous UiO-66 (HP-UiO-66) was rationally prepared using sodium acetate and polyvinylpyrrolidone as auxiliaries. The defective structure of HP-UiO-66 was confirmed by various characterization methods. Compared with the pristine UiO-66, HP-UiO-66 possessed more accessible Zr sites to load aptamers. The electrochemical aptasensor based on HP-UiO-66 was used to detect ampicillin (AMP) as an analytical model with a low limit of detection (3.0 fg mL−1) by differential pulse voltammetry. This fabricated aptasensor exhibited good reproducibility, stability, and specificity toward AMP. The sensor could be used to quantitatively detect AMP even in real samples (tap water and milk).

Comparative Analysis of QCM and Electrochemical Aptasensors for SARS-CoV-2 Detection.

The rapid and accurate detection of SARS-CoV-2, particularly its spike receptor-binding domain (S-RBD), was crucial for managing the COVID-19 pandemic. This study presents the development and optimization of two types of aptasensors: quartz crystal microbalance (QCM) and electrochemical sensors, both employing thiol-modified DNA aptamers for S-RBD detection. The QCM aptasensor demonstrated exceptional sensitivity, achieved by optimizing aptamer concentration, buffer composition, and pre-treatment conditions, with a limit of detection (LOD) of 0.07 pg/mL and a linear range from 1 pg/mL to 0.1 µg/mL, and a significant frequency change was observed upon target binding. The electrochemical aptasensor, designed for rapid and efficient preparation, utilized a one-step modification process that reduced the preparation time to 2 h while maintaining high sensitivity and specificity. Electrochemical impedance spectroscopy (EIS) enabled the detection of S-RBD concentrations as low as 132 ng/mL. Both sensors exhibited high specificity, with negligible non-specific interactions observed in the presence of competing proteins. Additionally, the QCM aptasensor's functionality and stability were verified in biological fluids, indicating its potential for real-world applications. This study highlights the comparative advantages of QCM and electrochemical aptasensors in terms of preparation time, sensitivity, and specificity, offering valuable insights for the development of rapid, sensitive, and specific diagnostic tools for the detection of SARS-CoV-2 and other viruses.

Facile and controllable hybrid-nanoengineering of MWCNTs/Au@ZIF-8 and AuPt@CeO2 based sandwich electrochemical aptasensor for AFB1 determination in foods and herbs

Herein, a sandwich electrochemical sensing strategy for aflatoxin b1 (AFB1) detection based on hybrid-nanoengineering was presented. First, Au nanoparticle was doped into zeolitic imidazolate framework-8 (ZIF-8) to form Au@ZIF-8 by in-situ growth method, followed by multi-walled carbon nanotubes (MWCNTs) addition to synthesize MWCNTs/Au@ZIF-8 via self-assembly. The structural “confinement effect” of ZIF-8 afforded a microenvironment for Au nanoparticles and SMCNTs in a certain spatial region, giving MWCNTs/Au@ZIF-8 excellent electrochemical property as the substrate material. In addition, Au-Pt bimetallic nanoparticle, which exhibited excellent stability and catalytic activity was loaded on the hollow cerium oxide (CeO2) to form AuPt@CeO2 nanoparticle through one-step aqueous phase reduction. Owning to its high surface-to-volume ratio, satisfied electron transfer efficiency and biocompatibility, massive toluidine blue (TB) and AFB1 antibody (Ab) could be modified on the AuPt@CeO2 to form AuPt@CeO2-Ab-TB, which acted as signal tag for the ultrasensitive assay of AFB1. The proposed electrochemical sensing system exhibited wide detection range (2 × 10-5 − 20 ng/mL) and low detection limit (2.13 fg/mL), which has been successfully applied to AFB1 determination in four real samples. The hybrid nanoengineering presented in this work is an active attempt to prepare high-performance substrate material and signal tag, which provides a new insight for the development of highly sensitive and specific electrochemical sensing systems.

Construction of a Heterostructure Composite of Fe-Doped CeO₂ With Buckypaper: Integration as an Electrochemical Aptasensing Platform for Ochratoxin A Detection

Construction of a Heterostructure Composite of Fe-Doped CeO₂ With Buckypaper: Integration as an Electrochemical Aptasensing Platform for Ochratoxin A Detection

Disposable electrochemical aptasensor with mesoporous carbon spheres modified screen-printed dual-working electrodes for Pb2+ and Hg2+ detection

Electrochemical biosensing combined with screen-printed paper technology provides an effective strategy for the quantitative analysis of metal ions, particularly suitable for in-situ determination. In this study, a disposable, economical and user-friendly screen-printed dual-working electrodes (SPDEs) aptasensor system for the determination of lead ions (Pb2+) and mercury ions (Hg2+) was described. The sensor system consists of dual-working electrodes, a counter electrode and a reference electrode fabricated using screen printing technology. The aptasensor demonstrates high sensitivity and selectivity towards Pb2+ and Hg2+, owing to the larger specific surface area (1334.7 m2/g) and excellent conductivity of mesoporous carbon nanoparticles (MCNs). Additionally, the specific recognition ability and high affinity of G-quadruplex and T-Hg2+-T structures for Pb2+ and Hg2+, respectively, further enhance the performance. Under optimized experimental conditions, the aptasensor achieves a low limit of detection (LOD) of 0.088 ng/mL for Pb2+ and 0.0007 ng/mL for Hg2+ (S/N = 3), a wide linear range (0.1–1000.0 ng/mL for Pb2+ and 0.001–100.0 ng/mL for Hg2+), and high accuracy (relative standard deviation, RSD≤10.0 %) and selectivity. Importantly, the SPDEs aptasensor can detect Pb2+ and Hg2+ in a sample without competition and interference, achieving accuracy comparable to that of commercial inductively coupled plasma mass spectrometry (ICP-MS). These advantages make the SPDEs aptasensor highly suitable for practical applications in environmental monitoring, food safety, and clinical diagnosis.

Catalytic hairpin assembly-driven DNA walker to develop a label-free electrochemical aptasensor for antibiotic detection.

Anelectrochemical aptasensor was developed by utilizing a DNA walker driven by catalytic hairpin assembly (CHA) with kanamycin as the model analyte. Kanamycin bound to the aptamer, causesthe release of DNA walker, triggers the CHA reaction, leadsto the cyclic movement of the walker's long arm, and resultsin cascade amplification of thesignal. The guanine-rich sequences of the double-stranded products produced by CHA were folded to form G-quadruplex structures, with electrochemical active molecules Hemin embedded, formsG-quadruplex/Hemin complexes in situ on the electrode surface, thereby achieving sensitive, efficient, and label-free detection of kanamycin with a limit of detection (LOD) of 0.27pM (S/N = 3). Meaningfully, the aptasensor demonstrated high sensitivity and reliability in the detection of kanamycin in milk and livestock wastewater samples, suggesting that it has great potential for application in detecting antibiotics in food products and water samples from the environment.

Plasmon-enhanced fluorescence and electrochemical aptasensor for SARS-CoV-2 Spike protein detection

In this work, we combined plasmon-enhanced fluorescence and electrochemical (PEF-EC) transduction mechanisms to realize a highly sensitive dual-transducer aptasensor. To implement two traducers in one biosensor, a novel large-scale nanoimprint lithography process was introduced to fabricate gold nanopit arrays (AuNpA) with unique fringe structures. Light transmitting through the AuNpA samples exhibited a surface plasmon polariton peak overlapping with the excitation peak of the C7 aptamer-associated fluorophore methylene blue (MB). We observed a five and seven-times higher average fluorescence intensity over the AuNpA and fringe structure, respectively, in comparison to a plane Au film. Furthermore, the MB fluorophore was simultaneously utilized as a redox probe for electrochemical investigations and is described here as a dual transduction label for the first time. The novel dual transducer system was deployed for the detection of SARS-CoV-2 Spike protein via a C7 aptamer in combination with a strand displacement protocol. The PEF transducer exhibited a detection range from 1 fg/mL to 10 ng/mL with a detection limit of 0.07 fg/mL, while the EC traducer showed an extended dynamic range from 1 fg/mL to 100 ng/mL with a detection limit of 0.15 fg/mL. This work provides insights into an easy-to-perform, large-scale fabrication process for nanostructures enabling plasmon-enhanced fluorescence, and the development of an advanced but universal aptasensor platform.

A dual-targeted electrochemical aptasensor for neuroblastoma-related microRNAs detection

Neuroblastoma (NB) is a significant pediatric cancer associated with high mortality rates, demanding innovative and appropriate approaches for its accurate detection. This paper described the design of a dual-target electrochemical aptasensor capable of simultaneously detecting neuroblastoma-associated microRNAs (miRNA-181 and miRNA-184) with exceptional sensitivity. Screen-printed carbon electrodes (SPCEs) were utilized with gold nanorods (AuNRs), and aptamers functionalized gold nanoparticles (AuNPs) to improve sensitivity, specificity, and portable detection ability. The detection method employed in this study includes differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Our aptasensor exhibited remarkable limits of detections (LODs) of 5.10 aM for miRNA-181 and 9.39 aM for miRNA-184, respectively, along with a broad linear range spanning from 0.1 fM to 100 pM for both miRNAs. The practical significance of neuroblastoma diagnosis was shown through the validation of serum samples and comparison with quantitative polymerase chain reaction (qPCR). Our electrochemical aptasensor is user-friendly, easy to engineer, and offers a promising approach for accurately and selectively detecting important miRNA biomarkers in cancer screening and diagnosis, showing potential application in various clinical scenarios.

Electrochemical Aptasensor with Antifouling Properties for Label-Free Detection of Oxytetracycline.

Oxytetracycline (OTC) is a widely employed antibiotic in veterinary treatment and in the prevention of infections, potentially leaving residues in animal-derived food products, such as milk, that are consumed by humans. Given the detrimental effects of prolonged human exposure to antibiotics, it has become imperative to develop precise and sensitive methods for monitoring the presence of OTC in food. Herein, we describe the development and results of a preliminary label-free electrochemical aptasensor with antifouling properties designed to detect OTC in milk samples. The sensor was realized by modifying a gold screen-printed electrode with α-lipoic acid-NHS and an amine-terminated aptamer. Different electrochemical techniques were used to study the steps of the fabrication process and to quantify OTC in the presence of the Fe(CN)64-/Fe(CN)63- redox couple The detectable range of concentrations satisfy the maximum residue limits set by the European Union, with an limit of detection (LOD) of 14 ng/mL in phosphate buffer (BP) and 10 ng/mL in the milk matrix, and a dynamic range of up to 500 ng/mL This study is a steppingstone towards the implementation of a sensitive monitoring method for OTC in dairy products.

A novel electrochemical aptasensor based on NrGO-H-Mn3O4 NPs integrated CRISPR/Cas12a system for ultrasensitive low-density lipoprotein determination.

Atherosclerosis cardiovascular disease (ASCVD) has become one of the leading death causes in humans. Low-density lipoprotein (LDL) is an important biomarker for assessing ASCVD risk level. Thus, monitoring LDL levels can be an important means for early diagnosis of ASCVD. Herein, a novel electrochemical aptasensor for determination LDL was designed based on nitrogen-doped reduced graphene oxide-hemin-manganese oxide nanoparticles (NrGO-H-Mn3O4 NPs) integrated with clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR/Cas12a) system. NrGO-H-Mn3O4 NPs not onlyhave alarge surface area and remarkable enhancedelectrical conductivity but also the interconversion of different valence states of iron in hemin can provide an electrical signal. Nonspecific single-stranded DNA (ssDNA) was bound to NrGO-H-Mn3O4 NPs to form a signaling probe and was immobilized on the electrode surface. The CRISPR/Cas12a system has excellent trans-cleavage activity, which can be used to cleave ssDNA, thus detaching the NrGO-H-Mn3O4 NPs from the sensing interface and attenuating the electrical signal. Significant signal change triggered by the target was ultimately obtained, thus achieving sensitive detection of the LDL in range from 0.005 to 1000.0nM with the detection limit of 0.005nM. The proposed sensor exhibited good stability, selectivity, and stability and achieved reliable detection of LDL in serum samples, demonstrating its promising application prospects for the diagnostic application of LDL.

Label-free SELEX of aptamers for ultra-sensitive electrochemical aptasensor detection of amanitin in wild mushrooms

BackgroundMushroom poisoning poses a significant global health concern, with high morbidity and mortality rates. The primary lethal toxins responsible for this condition are alpha-amanitin (ɑ-AMA) and beta-amanitin (β-AMA). As a promising bio-recognition molecules in biosensors, aptamers, have been broadly used in the field of food detection. However, the current SELEX-based methods for screening aptamers for structurally similar small molecules were limited by the labelling or salt ion induction. In this study, we aimed to develop a novel label-free SELEX strategy for the screening of aptamers with high affinity and constructed new aptasensors for the detection of ɑ-AMA and β-AMA. ResultsA novel label-free SELEX strategy based on the positively charged gold nanoparticles (AuNPs) was proposed to simultaneous screening of aptamers for ɑ-AMA and β-AMA. Only 18 rounds of SELEX were required to obtain new aptamers. The candidate aptamers were analyzed by colloidal gold assay, and the sequences of ɑ-30 and β-37 displayed great affinity with Kd values of 22.26 nM and 23.32 nM, respectively, without interference from botanical toxins. Notably, the truncated aptamers ɑ-30-2 (50 bp) and β-37-2 (57 bp) exhibited higher affinity than their original counterpart (79 bp). Subsequently, the selected aptamers were utilized to construct recognition probes for electrochemical aptasensors based on hairpin cyclic cleavage of substrates by Cu2+ dependent DNAzyme and Exo I-triggered recycling cascades. The detection platform showed excellent analytical performance with limits of detection as low as 4.57 pg/mL (ɑ-AMA) and 8.49 pg/mL (β-AMA). Moreover, the aptasensors exhibited superior performance in mushroom and urine samples. SignificanceThis work developed a simple and efficient label-free SELEX method for screening new aptamers for ɑ-AMA and β-AMA, which employed the positively charged AuNPs as the screening medium, without the need for chemical labelling of libraries or induction of salt ions. Furthermore, two novel electrochemical aptasensors were developed based on our newly obtained aptamers, which offer the new biosensing tool for ultrasensitive detection of the AMA poisoning, showing great potential in practical applications.

Design of a facile and highly sensitive electrochemical impedimetric aptasensor for detection of 17β-estradiol

In this work, a facile, rapid and label-free electrochemical (EC) impedimetric aptasensor was developed for detecting 17β-estradiol (E2) based on gold particles (Au NPs) signal amplification. The aptamer was used as the recognition and capture unit to label on the surface of Au NPs. The anti-E2 aptamer labelled Au NPs (apt-Au NPs) were then anchored on the complementary DNA-immobilized gold electrode surface by hybridization. Owing to the large surface, good biocompatibility and high conductivity, Au NPs can not only provide a wonderful microenvironment for aptamer, but also play a signal amplification role, improve analytical performance of the EC sensing platform. Due to the high affinity and specificity of aptamer to E2, the formed aptamer-E2 complexes with poor conductivity increase the electron transfer resistance (Ret). Quantitative analysis for E2 was performed based on the correlation between the change of the Ret and E2 concentrations. The developed sensor exhibited excellent analytical performance with high sensitivity, selectivity and good stability for the detection of E2. The detection limit for E2 is 0.053 pM, with a linear response ranging 0.1–200 pM. Meanwhile, its application in real environmental samples was evaluated and satisfying results were achieved.

DNA Aptamer Integrated Hydrogel Nanocomposites on Screen Printed Gold Electrodes for Point-of-Care Detection of Testosterone in Human Serum.

This work describes the development and evaluation of a novel electrochemical aptasensor for testosterone detection. The sensor utilizes a specifically designed DNA immobilized on a screen-printed gold electrode (SPGE) modified with a conductive hydrogel and gold nanoparticles (HG/NP) composite. The aptasensor exhibited a dose-dependent response to testosterone (0.05 to 50 ng/mL) with a detection limit of 0.14 ng/mL and a good sensitivity of 0.23 μA ng-1 mL cm-2. The sensor displayed excellent selectivity towards testosterone compared to structurally similar steroid hormones. Importantly, the incorporation of HG/NP not only improved the sensor's conductivity but also acted as an antifouling layer, minimizing signal interference from non-specific biomolecule interactions in complex biological samples like human serum. The results obtained from the aptasensor showed good correlation with a standard ELISA method, demonstrating its effectiveness in real-world scenarios.

Aptamer and aptasensor technology for diagnosis of infectious diseases: A mini review

BackgroundAptamers are not so new a concept, however, it is scarcely discussed by medical fraternity. Aptamers are potent, new identification molecules set to rope in a new technique in the diagnostic arena. Aptamers have started almost a revolution in diagnostic assays since their discovery in the 90s. (Radu S. Current and previous disease outbreaks around the world, U.S. News & World Report. 2020 Mar 13 [cited 2024 Jun 17]. Available from: https://www.usnews.com/news/best-countries/slideshows/20-pandemic-and-epidemic-diseases-according-to-who) provides an overview of pandemics and epidemics as reported by the WHO. It is interesting to note that several endemic and epidemic diseases viz. Chikungunya, Cholera, Crimean-Congo haemorrhagic fever, Ebola virus disease, Hendra virus infection, Influenza, Lassa fever, Marburg virus disease, Meningitis, MERS-CoV (Middle East Respiratory Syndrome Corona Virus), Monkeypox, Nipah virus infection, Novel coronavirus, Plague, Rift Valley fever, SARS (Severe Acute Respiratory Syndrome), Smallpox, Tularaemia, Yellow fever, and Zika virus disease have been identified by the WHO and are being explored for applicability of aptamer technology in their identification. ObjectivesOne of the most important necessities to control epidemic or pandemic diseases is early diagnosis. However, the majority of the diagnostic tests for these diseases are available only in tertiary care centres. The objective of this review is to discuss the potential of aptamer technology to provide undemanding, simple, specific, sensitive, and cost-effective diagnostic assays that are useable in remote and field conditions. ContentHere, we discuss recent advances and approaches in aptamer and aptamer engineering useful in the diagnosis of infectious and non-infectious conditions. This review also discusses a few sensing discoveries which are a gift of advanced engineering and technology using optical and electrochemical aptasensors. It's still a long way to go, and we need to take into account the technological challenges being faced by aptamer-aptasensor technology.