Hairpin Aptamer Research Articles

Ultrasensitive electrochemical biosensor for bacteria detection based on Fe3O4@COF-AuNPs and trigging isothermal circular amplification

Foodborne bacterial infections pose serious threats to human health, leading to illnesses and even fatalities. The key to solve this problem is to develop fast and effective sensor for sensitive detection of bacteria. In this study, an electrochemical biosensor based on magnetic nanocomposite material Fe3O4@COF-AuNPs and trigging isothermal circular amplification (TICA) was constructed for detection of foodborne bacteria. In this design, a hairpin aptamer probe (HAP)-bound DNA monolayer was designed, serving as the substrate for the electrode to increase surface area. The Escherichia coli (E. coli) was selected as the target. The presence of E. coli caused the configuration change of the HAP and allowed the occurrence of TICA with the use of the Klenow Fragment polymerase (KF-Poly), thus the HAPs were away from the DNA monolayer. After adding the catalytic magnetic nanocomposite material Fe3O4@COF-AuNPs, it was immobilized on the DNA monolayer of the electrode through DNA complementary pairing and exhibited high electrocatalytic behavior towards the reduction of 4-nitrophenol. This enabled ultrasensitive detection of E. coli, with the detection limit (LOD) as low as 10 CFU/mL. The detection time was 1 h. This designed electrochemical sensor showed high selectivity, stability, and reproducibility, which offered a new avenue for detection of foodborne bacteria.

Advancing Alzheimer’s biomarker Detection: A hairpin Aptamer-Based competitive electrochemical biosensor for trace Amyloid beta 40 analysis

Accurate detection of Alzheimer’s disease (AD) core biomarkers in body fluid is critical for identifying latent asymptomatic AD patients and prediction AD progress. Amyloid beta 1–40 (Aβ-40) as one of the core biomarkers, is expected to be an early diagnostic indicator for AD. Here, we establish a competitive method based on hairpin-structured aptamer for rapid and highly sensitive detection of Aβ-40 using square wave voltammetry (SWV). A hairpin aptamer with redox-active moiety methylene blue (MB) are immobilized on a gold working electrode as a recognition element. In the detection, a short complementary oligonucleotide strand is first introduced to switch on the hairpin, whereafter the targets Aβ-40 join in and selectively react with the aptamer in a competitive manner with the complementary strand, leading to the changes in the proximity of MB to the electrode and thereby in the electrochemical signal. Various parameters such as aptamer concentration, complementary chain length, incubation time, and temperature were optimized to achieve maximum sensing performance. By introducing a complementary chain, the sensor significantly reduces the detection limit to 7.14 pg/mL, with a dynamic range of 20 pg/mL to 200 ng/mL. Additionally, high selectivity of the sensor for Aβ-40 monomer (Aβ-40 m) over other forms of Aβ peptides has been validated. The hairpin-mediated detection method enables rapid, facile and highly sensitive detection of Aβ-40 using low-cost screen-printed electrode (SPE) platform, providing a wide range of possibilities for the early diagnosis of AD in point-of-care technologies (POCT).

Nanoenzyme Hydrogel Film-Based Portable Point-of-Care Testing Platform for Double-Signal Visual Detection of PSA.

The development of the Point-of-Care Testing (POCT) platform that combines convenience and cost-effectiveness is crucial for enabling the visual detection of disease biomarkers. In this work, a POCT platform for the sensitive in situ detection of prostate specific antigen (PSA) with dual-signal output was constructed by functionalizing the Eppendorf (EP) tube. This was achieved through the modification of aptamer hairpin probes (AHPs) on the lid of the EP tube and the assembly of a nanoenzyme hydrogel film on its inner wall. The target could trigger the release of Ag+ by AHP and subsequently activate Ag+-dependent DNAzyme (Ag-DNAzyme). This would initiate the cleavage of the DNA-Au/Pt NP hydrogel network, leading to the release of Au/Pt NPs. The released Au/Pt NPs exhibit both peroxidase (POD)-like and catalase (CAT)-like activity to produce a colorimetric response and induce liquid flow under pressure. Therefore, the target can be measured visually and quantitatively through colorimetric analysis and the measurement of total dissolved solids (TDS) using a pressure-triggered liquid flow device integrated into the platform. The designed platform is distinguished by its simplicity, specificity, cost-effectiveness, and remarkable sensitivity. It allows for the visual detection of PSA within concentration ranges of 0.5-100 ng/L (colorimetric) and 3-100 ng/L (TDS reading), boasting detection limits as low as 0.15 ng/L (colorimetric) and 0.57 ng/L (TDS reading). The strategy of target-triggered nanoenzyme release significantly enhances sensitivity and provides a guiding approach for visual biomarker detection.

Noncompetitive Determination of Small Analytes by Sandwich-Type Lateral Flow Assay Based on an Aptamer Kissing Complex.

Here, we present the proof-of-concept of a lateral flow assay (LFA) that is capable of detecting small-molecule targets in a noncompetitive manner by deploying a sandwich-type format based on the aptamer kissing complex (AKC) strategy. A fluorescently labeled hairpin aptamer served as the signaling agent, while a specific RNA hairpin grafted onto the strip served as the capture element. The hairpin aptamer switched from an unfolded to a folded form in the presence of the target, resulting in kissing interactions between the loops of the reporter and the capture agents. This design triggered a target-dependent fluorescent signal at the test line. The AKC-based LFA was developed for the detection of adenosine, achieving a detection limit in the micromolar range. The assay revealed the presence of the same analyte in urine. The method also proved effective with another small molecule (theophylline). We believe that the AKC-based LFA approach could overcome many of the shortcomings associated with conventional signal-off methods and competitive processes.

Interferon gamma (IFN-γ)-sensitive TB aptasensor based on novel chitosan-indium nano-kesterite (χtCITS)-labeled DNA aptamer hairpin technology

There has been increasing interest in the use of biosensors for diagnosis of infectious diseases such as tuberculosis (TB) due to their simplicity, affordability, and potential for point-of-care application. The incorporation of aptamer molecules and nanomaterials in biosensor fabrication explores the advantages of high-binding affinity and low immunogenicity of aptamers as well as the high surface-to-volume ratio of nanomaterials, for increased aptasensor performance. In this work, we employed a novel microwave-synthesized copper indium tin sulfide (CITS) substituted-kesterite nanomaterial, together with a natural biopolymer (chitosan), for signal amplification and increased loading of aptamer molecules. Study of the optical properties of CITS nanomaterials showed strong absorption in the UV region characteristic of kesterite semiconductor nanomaterials. X-ray diffraction analysis confirmed the presence of the kesterite phase with average crystallite size of 6.188 nm. Fabrication of interferon-gamma (IFN-γ) TB aptasensor with a chitosan-CITS nanocomposite (χtCITS) increased the aptasensor's electrochemical properties by 77.5 % and improved aptamer loading by 73.7 %. The aptasensor showed excellent sensitivity to IFN-γ concentrations with limit of detection of 6885 fg/mL (405 fM) and linear range of 850–17000 fg/mL (50 – 1000 fM). The aptasensor also exhibited excellent storage and electrochemical stability, with good selectivity towards IFN-γ and possible real sample application.

One-step and label-free ratiometric fluorescence assay for the detection of plasma exosome towards cancer diagnosis

Exosomes are closely associated with tumor development and are regarded as viable biomarkers for cancer. Here, a ratiometric fluorescence method was proposed for the one-step and label-free detection of plasma exosomes. A bicolor streptavidin magnetic beads were specifically created with an immobilized Cy5-labeled hairpin aptamer for CD63 (Cy5-Apt) on its surface to identify exosome, and a green color SYBR Green I (SGI) embedded in the stem of Cy5-Apt to respond to exosomes. After exosome capture, the Cy5-Apt could undergo a conformational shift and release the encapsulated SGI, allowing exosome measurement based on the fluorescence ratio of Cy5 and SGI. The enrichment, separation and detection of exosomes in proposed method could be completed in one step (30 min), which is a significant improvement over previous method. Furthermore, the use of ratiometric fluorescence and magnetic separation allows for exosome enrichment and interference elimination from complex matrices, improving accuracy and sensitivity. Particularly, the assay could detect exosomes in plasma and has potential to distinguish lung cancer patients from healthy volunteers with an area under the receiver operator characteristic curve of 0.85. Besides, the study provided an efficient method for analyzing the various divisions of exosomes by merely modifying the aptamer, which holds great promise for point-of-care applications.

G-Quadruplex/Hemin-Mediated Polarity-Switchable and Photocurrent-Amplified System for Escherichia coli O157:H7 Detection.

The contamination of food by pathogens is a serious problem in global food safety, and current methods of detection are costly, time-consuming, and cumbersome. Therefore, it is necessary to develop rapid, portable, and sensitive assays for foodborne pathogens. In addition, assays for foodborne pathogens must be resistant to interference resulting from the complex food matrix to prevent false positives and negatives. In this study, hemin and reduced graphene oxide-MoS2 sheets (GMS) were used to design a near-infrared (NIR)-responsive photoelectrochemical (PEC) aptasensor with target-induced photocurrent polarity switching based on a hairpin aptamer (Hp) with a G-quadruplex motif. A ready-to-use analytical device was developed by immobilizing GMS on the surface of a commercial screen-printed electrode, followed by the attachment of the aptamer. In the presence of Escherichia coli O157:H7, the binding sites of Hp with the G-quadruplex motif were opened and exposed to hemin, leading to the formation of a G-quadruplex/hemin DNAzyme. Crucially, after binding to hemin, the charge transfer pathway of GMS changes, resulting in a switch of the photocurrent polarity. Further, G-quadruplex/hemin DNAzyme enhanced the cathodic photocurrent, and the proposed sensor exhibited a wide linear range ((25.0-1.0) × 107 CFU/mL), a low limit of detection (2.0 CFU/mL), and good anti-interference performance. These findings expand the applications of NIR-responsive PEC materials and provide versatile PEC methods for detecting biological analytes, especially for food safety testing.

Low-Cost Full-Range Detection of C-Reactive Protein in Clinical Samples by Aptamer Hairpin Probes and Coprecipitation of Silver Ions and Gold Nanoparticles.

C-reactive protein (CRP) levels can vary widely related to diverse disease contexts. However, expensive antibodies have impeded the clinical utility of antibody-based full-range CRP assays, especially in developing countries. Herein, we established a low-cost, antibody-free, 96-well plate-based full-range CRP detection method by combining gold nanoparticles (AuNPs), silver iodide (AgI), Eosin Y, and the aptamer hairpin probe (AHP) with Ag+-mediated cytosine-cytosine mismatches, that is, the Au@AgI/Eosin Y-AHP method. After binding the target CRP, the AHP released Ag+, which subsequently induced the aggregation of AuNPs on the surface of AgI colloids, resulting in a significant increase in the adsorption of Eosin Y on the surface of AuNPs. The changes in fluorescence intensity (FI) of Eosin Y in the supernate without and with CRP were proportional to the concentration of the CRP in the wide range of 0.01-40 ng/mL (r = 0.9969), and 96 samples can be detected in 96-well plates simultaneously by a microplate reader within 45 min. Remarkably, the CRP levels of 100 clinical samples achieved with the Au@AgI/Eosin Y-AHP had a good correlation with those obtained with the latex-enhanced immune turbidimetry assay (r = 0.986). Furthermore, the kit based on the Au@AgI/Eosin Y-AHP method costs only $8.1 for 100 tests. Therefore, the new method is beneficial for less developed areas where expensive assays are not affordable.

Rational design of hairpin aptamer using intrinsic disorder mechanism to enhance sensitivity of aptamer folding-based electrochemical sensor for tobramycin

Folding-based electrochemical sensors employing electrode-immobilized, structure-switching aptamers represent a promising platform for rapid and selective detection of a variety of target analytes. Sensitivity of this class of sensors usually needs, however, to be improved through re-engineering the parent aptamer probes to undergo large conformation change upon target binding. Here, we report the development of a sensitive folding-based electrochemical sensor for tobramycin by re-engineering a DNA hairpin aptamer based on the concept of intrinsic disorder. Through incorporating disordered poly-thymine base spacers of various lengths into the loop of the parent hairpin aptamer, structure-switching aptamers exhibiting large target binding-induced conformation change for high signal sensitivity were re-engineered and an optimal hairpin aptamer containing a disordered spacer of 24 thymine bases (probe T24) was obtained. Folding-based electrochemical sensor modified with the probe T24 is sensitive for tobramycin with a maximum percent signal change of 767.1% for signal-saturating target concentration compared to 54.7% obtained using the parent aptamer. This sensor is also selective enough and applicable to detection of tobramycin in real samples with satisfactory results. We believe that the intrinsic disorder-based concept for re-engineering of structure-switching aptamers will help guide further design and application of highly sensitive aptamer folding-based electrochemical sensors.

Electrochemical Detection of Tumor Cell-Derived Exosomes Based on Cyclic Enzyme Scission and Hybridization Chain Reaction Dual-Signal Amplification

Tumor cell-derived exosomes are considered a potential source of cancer biomarkers. Here, we developed an electrochemical sensing platform for the rapid and simple detection of exosomes, using the CCRF-CEM exosome as a model. The platform utilizes cyclic nicking enzyme cleavage and a hybridization chain reaction (HCR) for dual-signal amplification. A hairpin aptamer probe (HAP) containing an aptamer was designed for the assay. The specific binding between the aptamer and PTK7, present on the exosome surface, causes a conformational change in the HAP. This facilitates hybridization between the HAP and the linker DNA, which subsequently triggers cyclic cleavage of the nicking endonuclease towards the linker DNA. Therefore, exosome detection is transformed into DNA detection. By combining this approach with HCR signal amplification, we achieved high-sensitivity electrochemical detection of CCRF-CEM exosomes, down to 1.1 × 104 particles/mL. Importantly, this assay effectively detected tumor exosomes in complex biological fluids, demonstrating the potential for clinical diagnosis.

High electrochemical active Au-NP/2D zinc-metal organic frameworks heterostructure-based ECL sensor for the miRNA-522 detection in triple negative breast cancer

In this work, a novel electrochemiluminescence (ECL) sensor has been developed to detect the miRNA-522 in the tumor tissues of triple-negative breast cancer (TNBC) patients. Au NPs/Zn MOF heterostructure was obtained by in situ growth and used as novel luminescence probe. Firstly, zinc-metal organic framework nanosheets (Zn MOF NSs) were synthesized with Zn2+ as the central metal ion and 2-aminoterephthalic acid (NH2-BDC) as the ligand. 2D MOF nanosheets with ultra-thin layered structure and relatively large specific surface areas can enhance the catalytic activity in the ECL generation. Furthermore, the electron transfer capacity and the electrochemical active surface area of MOF were greatly improved by the growth of Au NPs. Therefore, Au NPs/Zn MOF heterostructure showed the significant electrochemical activity in the sensing process. In addition, the magnetic Fe3O4@SiO2@Au microspheres were used as capture units in the magnetic separation step. The magnetic spheres with hairpin aptamer H1 can capture target gene. Then the captured miRNA-522 triggered the target catalyzed hairpin assembly (CHA) sensing process and linked Au NPs/Zn MOF heterostructure. The concentration of miRNA-522 can be quantified by the ECL signal enhancement of the Au NPs/Zn MOF heterostructure. Due to the high catalytic activity of Au NPs/Zn MOF heterostructure and their unique structural and electrochemical properties, the prepared ECL sensor achieved high-sensitive detection of miRNA-522 in the range of 1 fM to 0.1 nM with the detection limit of 0.3 fM. This strategy can provide a potential alternative for miRNA detection in medical research and clinical diagnosis of triple negative breast cancer.

Target/aptamer binding-induced inhibition of enzyme activity for amplified electrochemical detection of Sonic Hedgehog protein

The high expression of Sonic Hedgehog (SHh) is responsible for the known biological activity of Hedgehog, which closely correlates with cancer metastasis, drug resistance and poor prognosis of hepatocellular carcinoma. In this study, with a new target/aptamer binding-induced inhibition of enzyme activity strategy, we report the construction of an amplified and sensitive electrochemical biosensor for detecting SHh. The analytes bind hairpin aptamer probes to form SHh/aptamer complexes, which inhibit cleavage of aptamers by exonuclease III via the steric hindrance effect not to yield the displacement strands that can displace the biotin-modified signal probes from the sensor electrode. The streptavidin-horseradish peroxidase (STV-HRP) subsequently bind the biotin-probes and are confined on electrode surface through biotin/streptavidin interactions to show highly enhanced catalytic currents with the presence of 3, 3′, 5, 5′-tetramethylbenzidine (TMB)/H2O2 for realizing SHh detection down to 3.34 pM with high selectivity. The interrogation of SHh in diluted serums by this sensor has also been demonstrated. With the verification for SHh assay and the advantages in terms of aptamer recognition, simplicity by electrochemical transduction and sensitivity by catalytic current signal amplification, our strategy can be a convenient aptamer-based approach for detecting different proteins for various biological studies and disease diagnosis purposes.

Self-Assembly of Multivalent Aptamer-Tethered DNA Monolayers Dedicated to a Fluorescence Polarization-Responsive Circular Isothermal Strand Displacement Amplification for Salmonella Assay.

Pathogenic bacteria are pathogens widely spread that are capable of causing mild to life-threatening diseases in human beings or other organisms. Rationally organizing the simple helical motif of double-stranded DNA (dsDNA) tiles into designed ensemble structures with architecturally defined collective properties could lead to promising biosensing applications for pathogen detection. In this work, we facilely engineered multivalent hairpin aptamer probe-tethered DNA monolayers (MHAP-DNA monolayers) and applied them to build a fluorescence polarization-responsive circular isothermal strand displacement amplification (FP-CSDA) for Salmonella assay. In this system, the MHAP-DNA monolayers were constructed based on a dsDNA tile-directed self-assembly. A FAM-labeled reporting probe (RPFAM) with an inherent low FP signal serves as the signaling unit. The presence of target Salmonella leads to the trapping of F RPFAM into the super DNA monolayers via a target-triggered CSDA to peel off the tethered hairpin-structured aptamer probes (HAPs) responsible for the binding of RPFAM. As a result, the FP signal of the FAM fluorophore can be remarkably amplified due to the recycling of target Salmonella and the capacity of structural DNA materials to strongly restrict the free rotation of the FAM fluorophore but without a fluorescence quenching effect. Experimental results demonstrate that the FP assay is able to detect Salmonella with a low limit of detection (LOD) of 7.2 × 100 CFU/mL and high specificity. As a proof-of-concept study, we envision our study using DNA nanoarchitecture as the foundation to modulate CSDA-based FP assays, promising to open up a new avenue for disease diagnosis, food safety detection, and biochemical studies.

Detection of saxitoxin by a SERS aptamer sensor based on enzyme cycle amplification technology.

Saxitoxin (STX) is a typical toxic guanidinium neurotoxin, one of the paralytic shellfish poisons (PSP), which poses a serious threat to human health. In this paper, a simple and sensitive SERS aptamer sensor (abbreviated as AuNP@4-NTP@SiO2) for the quantitative determination of STX was developed. Hairpin aptamers of saxitoxin are modified on magnetic beads and used as recognition elements. In the presence of STX, DNA ligase, and the rolling circle template (T1), a rolling circle amplification reaction was triggered to produce long single-stranded DNA containing repetitive sequences. The sequence can be hybridized with the SERS probe to realize the rapid detection of STX. Due to the inherent merits of its components, the obtained AuNP@4-NTP@SiO2 SERS aptamer sensor manifests excellent sensing performance for STX detection with a wide linear range from 2.0 × 10-10 mol L-1 to 5.0 × 10-4 mol L-1 and a lower detection limit of 1.2 × 10-11 mol L-1. This SERS sensor can provide a strategy for the micro-detection of other biological toxins by changing the aptamer sequence.

Dual-amplification colorimetric detection of bisphenol A based on catalytic hairpin assembly and DNAzyme-caused fragment self-assembly hybridization chain reaction.

An efficient and innovative strategy for colorimetric detection of bisphenol A (BPA) is shown here based on target-induced catalytic hairpin assembly (CHA) and DNAzyme-caused fragment self-assembly hybridization chain reaction (HCR). BPA can bind with its aptamer hairpin to trigger CHA, thus forming Y-shaped DNA nanostructures with an enzyme-strand (E-DNA) tail. Subsequently, the E-DNA can cyclically cleave the substrate hairpin, generating many fragments which can cause self-assembly HCR to form long strand DNA. Finally, the formed long strand DNA can hybridize with short single strand DNA on AuNPs, causing the color change of AuNPs from red to blue. Six important detection conditions of the proposed aptasensor were optimized. Under optimal conditions, the biosensor has high sensitivity for BPA detection at concentrations ranging from 0.8 pM to 500 pM and the detection limit is as low as 0.2 pM, providing a promising prospective ultrasensitive detection of BPA.

A biodegradable and cofactor self-sufficient aptazyme nanoprobe for amplified imaging of low-abundance protein in living cells

Despite the progress on the analysis of proteins either in vitro or in vivo, detection and imaging of low-abundance proteins in living cells still remains challenging. Herein, a novel biodegradable and cofactor self-sufficient DNAzyme nanoprobe has been deve-loped for catalytic imaging of protein in living cells with signal amplification capacity. This DNAzyme nanoprobe is constructed by assembling a DNAzyme subunit-containing aptamer hairpin (HP), another DNAzyme subunit strand (DS), and the molecular beacon (MB) substrate strand onto pH-sensitive ZnO@polydopamine nanorods (ZnO@PDA NRs) that work as DNAzyme cofactor suppliers. Such a nanoprobe can facilitate cellular uptake of DNA molecules and protection of them from nuclease degradation as well as release of them in cells by lysosomal acid-triggered dissolution of ZnO@PDA NRs into Zn2+ as DNAzyme cofactor. Upon recognition and binding with the intracellular protein target, the stem of HP is opened, after which the opened HP hybridizes with DS and generates activated DNAzymes. Each activated DNAzyme can catalyze the cleavage of many MB substrates through true enzymatic multiple turnovers, resulting in the separation of the quenched fluorophore/quencher pair labeled in MB and the generation of significantly amplified fluorescence. Using nucleolin (NCL) as a model protein, this nanoprobe enables the analysis of NCL with a detection limit of 1.8 pM, which are at least two orders of magnitude lower than that of non-catalytic imaging probe. Moreover, it could accurately distinguish tumor cells and normal cells by live cell NCL imaging. And the experimental results are also further verified by flow cytometry assays. The developed nanoprobe can be easily extended to detect other biomolecules by the change of their corresponding aptamer sequences, thus providing a promising tool for highly sensitive imaging of low-abundance biomolecules in living cells.

The binding model of adenosine-specific DNA aptamer: Umbrella sampling study

We report a novel model of the selective binding mechanism of adenosine-specific DNA aptamer. Our theoretical investigations of AMP (Adenosine monophosphate) dissociation from aptamer-AMP complexes reveals new details of aptamer molecular specificity and stabilisation factors. Umbrella sampling MD calculations using parmbsc1 force field shows that the disordered structure of the internal loop of the unbound aptamer hairpin has a characteristic packing of guanines, which prevents barrier-free penetration of ligands into the site cavity. Also, this disordered structure of the unbound aptamer has a network of hydrogen bonds stabilising the cavity near the target guanines within the binding sites during the whole binding process. We suggested that the first AMP molecule binds to the disordered structure of the site closest to the aptamer hairpin stem and spends some free energy on ordering of the internal loop. Then the second AMP molecule binds to the ordered site closest to the aptamer hairpin loop with a lower energy gain. As a result, the induced-fit binding model is the most applicable for this aptamer and does not contradict the modern experimental NMR and calorimetry data.

Colorimetric aptasensor for on-site detection of acetamiprid with hybridization chain reaction-assisted amplification and smartphone readout strategy

An instruments-free colorimetric assay is established for acetamiprid detection with hybridization chain reaction (HCR)-based amplification strategy. Hairpin probes help gold nanoparticles (AuNPs) to resist salt-induced aggregation through sticky ends. In the presence of acetamiprid, aptamer hairpin probe HP opens due to the specifical recognition, then triggers an explosive hybridization between hairpin probes HA-1/2 and numerous hairpin probes are consumed. With the depletion of hairpin probes, the protection weakens and AuNPs suffer from salt-induced aggregation, which results in a color change from pinkish purplish to blue-purple. Under optimal conditions, the assay exhibits high sensitivity for acetamiprid analysis with a detection limit at 1.74 μM. Meanwhile, the aptasensor displays excellent selectivity that only acetamiprid can trigger the HCR and achieve enhanced detection. Worthwhile, based on the color variation, a smartphone readout-based strategy is proposed for visual quantification of acetamiprid, and good practical performance is obtained in real agricultural samples. The above visual method is portable and economic that get rid of advanced instruments, and shows a promising application prospect for on-site detection of acetamiprid.

A dual-recognition-controlled electrochemical biosensor for accurate and sensitive detection of specific circulating tumor cells

Accurate and sensitive assay of specific circulating tumor cells (CTCs) is of importance for the diagnosis, treatment, and metastasis monitoring of cancer. Herein, we have proposed a dual-recognition-controlled electrochemical biosensor in this work for the detection of specific CTCs. To this sensor, two aptamer hairpin probes are designed to be able to separately bind to two adjacent proteins on the cell membrane to activate the associative toehold for strand displacement reaction, which will then trigger a dimer-like rolling cycle amplification reaction, and finally produce significantly amplified electrochemical signals for sensitive quantification of target CTCs. In our design, only the case that the two proteins are simultaneously expressed on the cell membrane can result in obvious signal responses, which may greatly improve the accuracy of CTCs analysis. The proposed biosensor can possess excellent selectivity to distinguish target cells from different cancer cells. Moreover, the combination of rolling cycle amplification and DNA nanostructure capture probes can effectively lower the detection limit to 3 cells mL−1. Notably, our biosensor can be applied to the assay of the target CTCs in the complex whole blood matrixes, verifying its strong stability and anti-interference. Thus, the as-proposed dual-recognition-controlled electrochemical biosensor may exhibit great promise in clinical cancer diagnosis and personalized medicine.

Luminous MoS2 nanosheet-based electrochemiluminescence biosensor with biomimetic vesicle for miRNA-210 detection

In this work, a novel electrochemiluminescence (ECL) sensor has been developed to detect miRNA-210 in the serum of triple negative breast cancer (TNBC) patients. The luminous MoS2 nanosheets were synthesized via the solvothermal method and served as ECL emitters for the first time. As a result, the ECL properties of as-prepared MoS2 nanosheets were significantly improved. Furthermore, the biomimetic magnetic vesicles were used as capture platform in the ECL sensing strategy. Due to the highly efficient fluidity and magnetic property, the biomimetic vesicles with hairpin aptamers can capture target gene in the serum. After magnetic separation, the captured miRNA-210 can trigger the target-catalyzed hairpin assembly (CHA) sensing process on the magnetic electrode and hybridize MoS2 nanosheets labeled probe DNA. The concentration of miRNA-210 can be quantified by the ECL enhancement of the MoS2 nanosheets. This approach has achieved the sensitive detection for miRNA-210 in a range from 1 fM to 100 pM with the detection limit of 0.3 fM. The luminous MoS2 nanosheets-based ECL sensing system with the biomimetic vesicles would provide a new pathway to explore 2D nanomaterials for developing a wide range of bioanalytical applications.