Signal Amplification Effect Research Articles

A signal amplification of near-infrared electrochemiluminescence immunosensor for SFTSV determination based on SiO2 photonic crystals nanomembrane

In this work, a novel near-infrared electrochemiluminescence signal amplification strategy was fabricated based on SiO2 photonic crystals nanomembrane as the ECL electrodes for sensitive and specific detection of SFTSV. The photonic crystal structure of the SiO2 nanomembrane has an excellent light scattering effect, which significantly increases the photon flux in the luminescent molecule and improves the excitation of the ECL molecule (CdTe@CdS QDs) by the re-radiation dipole electromagnetic field. In addition, the increase of the effective electroactive area of the photonic crystals modified electrode also prominently contributes to the ECL intensity. The results manifested that the SiO2 photonic crystals nanomembrane modified electrode constructed in this work can effectively improve the ECL intensity by about 7 times. Subsequently, a sandwich-type SFTSV immunosensor with a very low limit of detection of 0.0014 fg/mL was constructed. Due to the low cost and easy preparation of SiO2 photonic crystal solid-state electrodes, this work can not only effectively detect the SFTSV virus, but also provides an effective ECL signal amplification strategy.

A Target-Driven Self-Feedback Paper-Based Photoelectrochemical Sensing Platform for Ultrasensitive Detection of Ochratoxin A with an In2S3/WO3 Heterojunction Structure

Currently, developing versatile, easy-to-operate, and effective signal amplification strategies hold great promise in photoelectrochemical (PEC) biosensing. Herein, an ultrasensitive polyvinylpyrrolidone-treated In2S3/WO3 (In2S3-P/WO3)-functionalized paper-based PEC sensor was established for sensing ochratoxin A (OTA) based on a target-driven self-feedback (TDSF) mechanism enabled by a dual cycling tactic of PEC chemical-chemical (PECCC) redox and exonuclease III (Exo III)-assisted complementary DNA. The In2S3-P/WO3 heterojunction structure with 3D open-structure and regulable topology was initially in situ grown on Au nanoparticle-functionalized cellulose paper, which was served as a universal signal transducer to directly record photocurrent signals without complicated electrode modification, endowing the paper chip with admirable anti-interference ability and unexceptionable photoelectric conversion efficiency. With the assistance of Exo III-assisted cycling process, a trace amount of OTA could trigger substantial signal reporter ascorbic acid (AA) generated by the enzymatic catalysis of alkaline phosphatase, which could effectively provoke the PECCC redox cycling among the tris(2-carboxyethyl)phosphine acid, AA, and ferrocenecarboxylic at the In2S3-P/WO3 photoelectrode, initiating TDSF signal amplification. Based on the TDSF process induced by the Exo III-assisted recycling and PECCC redox cycling strategy, the developed paper-based PEC biosensor realized ultrasensitive determination of OTA with persuasive selectivity, high stability, and excellent reproducibility. It is believed that the proposed paper-based PEC sensing platform exhibited enormous potential for the detection of other targets in bioanalysis and clinical diagnosis.

Sensitive and direct electrochemical detection of bisphenol S based on 1T&2H-MoS2/CNTs-NH2 nanocomposites

A voltammetric sensor was constructed for ultra-trace BPS detection based on the signal amplification effect of 1T&2H-MoS2 and CNTs-NH2 composites.

A switchable electrochemical hairpin-aptasensor for ochratoxin A detection based on the double signal amplification effect of gold nanospheres

An OTA electrochemical sensor based on h-DNA and the double effect of gold nanospheres that can be applied for actual sample detection.

Sandwich method-based sensitivity enhancement of Ω-shaped fiber optic LSPR for time-flexible bacterial detection

The development of rapid and sensitive detection methods for pathogenic bacteria is crucial for the therapy and prevention of related diseases. However, the rapid and ultrasensitive assays are difficult to be realized simultaneously. To solve the problem, a sandwich method based on Ω-shaped fiber optic localized surface resonance (Ω-FOLSPR) was constructed, where poly adenine-tailed aptamer (PolyA-apt) and SH modified gold nanoparticles tags (AuNPs tags) were chosen as the capturing aptamer and amplifying tags, respectively. The small AuNPs were modified on the surface of fiber-optic (FO) rapidly, which saved the preparation time. Then, the PolyA-apt was modified on the AuNPs surface to capture the bacteria effectively due to its ability to adjust the density and conformation of aptamer on the AuNPs surface. Finally, the large AuNPs tags were used to generate intense signal enhancement. It is found that the sandwich method enables the unique characteristic of a time-dependent sensitivity enhancement. Specifically, the LOD of 108.0 CFU/mL and 7.4 CFU/mL was achieved with the analysis time of 10 min and 100 min, respectively. Besides, the Ω-FOLSPR sensor exhibits excellent selectivity against the other bacteria and good performance for detecting the spiked and natural samples. This sandwich method provides a time-flexible strategy due to the combination of effective signal amplification and real-time analysis for bacterial detection, displaying great potential for practical bacterial detection.

A magnetic separation-assisted cascade hybridization chain reaction amplification strategy for sensitive detection of flap endonuclease 1

Sensitive and accurate detection of flap endonuclease 1 (FEN1) is essential to understand its roles in DNA replication and repair and explore its functions in tumor diagnosis and prognosis. Here, we designed a magnetic separation-assisted cascade hybridization chain reaction (HCR) amplification strategy for enzyme-free and sensitive detection of FEN1. Firstly, the forked dsDNAs contained the recognition site (5′ overhanging DNA flap) of FEN1 were modified on magbeads surface to form magnetic recognition probes. By the recognition and catalytic cleavage of FEN1, the 5′ flaps were removed into solution and then purified by magnetic separation. Then, the obtained flaps served as triggers to activate the linear-assembly of hairpin probes H1 and H2 (primary HCR), forming nicked double helices with split fragments. Finally, the fragments acted as new triggers to activate the branched-assembly of hairpin probes H3 and H4 (secondary HCR), causing fluorophore/quencher labeled on H3 far away and fluorescence recovery. Benefiting from the purification of triggers by magnetic separation and the signal amplification effect of cascade HCR, this strategy achieved a low detection limit of 3.6 × 10−5 U μL−1 for FEN1, and enabled the assay of it in actual samples. The results demonstrate that our strategy will provide a potent approach for sensitive detection of FEN1 in early tumor diagnosis and prognosis.

In situ hand-in-hand DNA tile assembly and CRISPR/Cas12a-based ultrasensitive detection for leukemia diagnosis

CRISPR/Cas technology has attracted increasing attention for use in accurate and sensitive nucleic acid detection, and has innovated the field of molecular diagnostics. Because it is programmability and shows high fidelity and effective signal amplification capabilities. Specifically, Cas12a has been evaluated through in-depth research because it triggers collateral trans-cleavage activity for the target. Here, we report a novel CRISPR/Cas12a fluorescent biosensor by in situ hand-in-hand DNA tile assembly for leukemia diagnosis, and it allows an active isothermal amplification for multi-channel collection of CRISPR/cas12a transducers for achieving three-stage signal amplification. When the target gene concentration was in a range from 1 fM to 50 nM, the fluorescence intensity exhibited an excellent linear relationship with the logarithm of the target gene concentration, and the detection limit reached 1 fM. Furthermore, good detection performance can still be maintained in actual sample analysis. The CRISPR/Cas biosensor for leukemia diagnosis is more advanced than the traditional CRISPR/Cas detection systems, which can be used in cancer gene analyses, kinship judgment, environmental monitoring, and so on.

Simultaneous detection of dual biomarkers using hierarchical MoS2 nanostructuring and nano-signal amplification-based electrochemical aptasensor toward accurate diagnosis of prostate cancer

Accurate and reliable quantification of tumor biomarkers in clinical samples is of vital importance for early stage diagnosis and treatment of cancer. However, a poor specificity of prostate specific antigen (PSA) testing alone fostering overdetection and overtreatment, remains a great controversy in prostate cancer (PCa) screening. Here we report an electrochemical aptasensor using hierarchical MoS2 nanostructuring and SiO2 nano-signal amplification for simultaneous detection of dual PCa biomarkers, PSA and sarcosine, to enhance the diagnostic performance of PCa. In this strategy, hierarchical flower-like MoS2 nanostructures as functional interface accelerated intermolecular accessibility and improved DNA hybridization efficiency. Moreover, the spherical SiO2 nanoprobe that conjugated with both electroactive tags and DNA probes, allowed effective electrochemical signal amplification. By deliberately designing different hybridization modes, we individually implemented the optimization of PSA and sarcosine sensing system. Based on this, simultaneous determination of PSA and sarcosine was achieved, with limit of detection (LOD) down to 2.5 fg/mL and 14.4 fg/mL, respectively, as well as excellent selectivity. More importantly, using this approach, we could directly differentiate cancer patients with healthy ones for clinical serum samples. The ultrasensitive biosensor provides single-step analysis with simple operation and a small sample volume (∼12 μL), shedding new light on accurate diagnosis and early-detection of cancer in clinical applications.

Sandwich-type photoelectrochemical immunosensor for procalcitonin detection based on Mn2+ doped CdS sensitized Bi2WO6 and signal amplification of NaYF4:Yb, Tm upconversion nanomaterial

In this paper, we constructed a sandwich-type photoelectrochemical (PEC) immunosensor for the quantitative detection of procalcitonin (PCT) based on the sensitization of Mn2+ doped CdS (CdS:Mn) nanocomposites to Bi2WO6 and the signal amplification effect of an upconversion material NaYF4:Yb,Tm. Bi2WO6 was synthesized with a three-dimensional flowered structure. CdS:Mn reduced the recombination of photogenerated carriers, and significantly improved photocurrent response. Lanthanide-doped upconversion nanomaterials were used as the label of secondary antibody. NaYF4:Yb,Tm have two functions, not only connected with the secondary antibody, but also can further amplify the photocurrent response. The proposed immunosensor for detecting PCT provided a desired linear range of 0.5 pg mL−1-100 ng mL−1 and a detection limit of 0.13 pg mL−1 under optimal experimental conditions. Besides, the PEC immunosensor demonstrated good reproducibility, specificity and stability. The results of determination of PCT in real human serum samples were satisfactory. Thus, the immunosensor may be applied in the clinical diagnosis of PCT and other biomarkers.

Ultrasensitive prostate specific antigen monitoring based on electrochemiluminescent immunesystem with synergistic signal amplification effect of resonance energy transfer coupling with K2S2O8-H2O2 dual coreactants

• The synergistic effect of coupling dual coreactants with the surface plasmon resonance of AgNPs enhancement was used as an effective signal amplification strategy. • OH • arising in the process was able to induce the decomposition of S 2 O 8 2− into SO 4 • − resulting in a vigorous ECL enhancement. • The amplified ECL immunosensor was fabricated for highly-sensitive detection of PSA. • The sensing system provides an alternative charming method for other biomolecules assays. In this work, an effective signal amplification strategy was used for the fabrication of the ultrasensitive sandwich-typed electrochemiluminescence (ECL) immunosensing platform. The combining of K 2 S 2 O 8 -H 2 O 2 dual coreactants with the localized surface plasmon resonance of silver nanoparticles (AgNPs) resulted in the remarkable increase of ECL response. Noticeably, because of the perfect overlap between the plasmon spectra of AgNPs and the ECL emission spectra of graphitic carbon nitride quantum dots (g-CNQDs), a plasmon-enhanced ECL phenomenon was proved in g-CNQDs/AgNPs system result from high efficient energy transfer. Besides, the ECL signal of the ECL immunosensor was greatly improved in the presence of K 2 S 2 O 8 -H 2 O 2 dual coreactants. Consequently, this synergistic amplified ECL sensing platform was applied for the specific prostate specific antigen (PSA) analysis. Under the optimum conditions, the immunoassay strategy demonstrated excellent sensing performance with a wide linear concentration of PSA from 1.0 × 10 − 5 ng mL −1 to 500 ng mL −1 and a detection limit of 0.0046 pg mL −1 (S/N = 3). Considering the favorable analytical feedback in actual serum samples assay, the developed strategy indicated an enormous potential for biomarkers analysis.

Pressure-Based Immunoassays with Versatile Electronic Sensors for Carcinoembryonic Antigen Detection.

Pressure-based immunoassays have been studied for point-of-care testing for which increasing the sensitivity is still a challenge. In this study, we described an enhanced pressure-based immunoassay with a versatile electronic sensor for the sensitive biological analysis. The versatile electronic sensor had multifunctional sensing capabilities with temperature and pressure recording. Magnetic bead-modified capture antibody and platinum nanoparticle-labeled detection antibody were used as the biorecognition element of the target carcinoembryonic antigen (CEA) (as a model analyte) and would form a sandwich-type immune complex with CEA. After simple magnetic separation, this complex was transferred into the detection chamber, which contained both hydrogen peroxide (H2O2) and 3,3',5,5'-tetramethylbenzidine (TMB). With the catalytic ability of PtNPs, the "H2O2-TMB-PtNPs" system was catalyzed to generate a large amount of oxygen (O2) and photothermal agent of oxidizer TMB (ox-TMB). Meanwhile, in a sealed chamber, further irradiation with an 808 nm near-infrared laser led to a triple-step signal amplification strategy of pressure increase, temperature increase, and gas thermal expansion to receive a strong electrical signal through the electronic sensor in real time. Thus, the amplified electrical signal from the electronic sensor could reveal the target concentration. In addition, we also verified that the synergistic system with two physical quantities had a lower limit of detection and a wider detection range compared to the detection system with a single physical quantity. In general, this immunoassay not only helped in exploring an effective signal amplification pathway but also offered an opportunity for the development of versatile electronic sensors in point-of-care settings.

Mass Spectrometric Biosensing: A Powerful Approach for Multiplexed Analysis of Clinical Biomolecules.

Rapid and sensitive detection of clinical biomolecules in a multiplexed fashion is of great importance for accurate diagnosis of diseases. Mass spectrometric (MS) approaches are exceptionally suitable for clinical analysis due to its high throughput, high sensitivity, and reliable qualitative and quantitative capabilities. To break through the bottleneck of MS technique for detecting high-molecular-weight substances with low ionization efficiency, the concept of mass spectrometric biosensing has been put forward by adopting mass spectrometric chips to recognize the targets and mass spectrometry to detect the signals switched by the recognition. In this review, the principle of mass spectrometric sensing, the construction of different mass tags used for biosensing, and the typical combination mode of mass spectrometric imaging (MSI) technique are summarized. Future perspectives including the design of portable matching platforms, exploitation of novel mass tags, development of effective signal amplification strategies, and standardization of MSI methodologies are proposed to promote the advancements and practical applications of mass spectrometric biosensing.

Enzyme-free sandwich-type electrochemical immunosensor for CEA detection based on the cooperation of an Ag/g-C3N4-modified electrode and Au@SiO2/Cu2O with core-shell structure

Effective signal amplification is a prerequisite for electrochemical immunosensors to achieve ultra-sensitive detection. In this work, we prepared a sandwich-type electrochemical immunosensor for the quantitative detection of carcinoembryonic antigen (CEA). As a base platform, Ag NPs modified aminated two-dimensional nitrogen carbide nanosheets (Ag/g-C3N4) have good biocompatibility and conductivity. In addition, with the layered structure of Au@SiO2/Cu2O as the signal label, the response current value of H2O2 was monitored by the Amperometric i-t Curve (i-t), so as to realize the accurate measurement of CEA. The presence of SiO2 nanoframes not only reduces the agglomeration of Au NPs and Cu2O but also provides good biocompatibility to facilitate the connection of secondary antibodies. Finally, we also verified the signal amplification mechanism of the immunosensor through XPS and other means, and calculated the kinetic parameters of the signal tag, which proved the good peroxidase-like activity of Au@SiO2/Cu2O. Under the best test conditions, the prepared immunosensor has a detection range from 0.01 pg/mL to 80 ng/mL, and the detection limit is as low as 0.0038 pg/mL. The results show that the immunosensor has good analytical performance and it can provide a new method for the clinical diagnosis of CEA.

A SERS-LFA biosensor combined with aptamer recognition for simultaneous detection of thrombin and PDGF-BB in prostate cancer plasma

An innovative surface-enhanced Raman spectroscopy and lateral flow assay (SERS-LFA) biosensor combined with aptamer recognition had been developed for the convenient, rapid, sensitive and accurate detection of thrombin and platelet-derived growth factor-BB (PDGF-BB) associated with prostate cancer simultaneously. During the biosensor operation, thrombin and PDGF-BB in the sample were recognized and combined by thiol-modified aptamers immobilized on Au–Ag hollow nanoparticles (Au–Ag HNPs) surface and biotinylated aptamers immobilized on the test lines of the biosensor. Thus, thrombin and PDGF-BB were simultaneously captured between detection aptamers and capture aptamers in a sandwich structure. Finite difference time domain simulation confirmed that ‘hot spots’ appeared at the gaps of Au–Ag HNPs dimer in the enhanced electromagnetic field compared to that of a single Au–Ag HNP, indicating that the aggregated Au–Ag HNPs owned a good SERS signal amplification effect. The detection limits of thrombin and PDGF-BB in human plasma were as low as 4.837 pg ml−1 and 3.802 pg ml−1, respectively. Moreover, the accuracy of the biosensor which was applied to detect thrombin and PDGF-BB in prostate cancer plasma had been verified. This designed biosensor had broad application prospects in the clinical diagnosis of prostate cancer.

Photo-gated and self-powered three-dimensional DNA motors with boosted biostability for exceptionally precise and efficient tracing of intracellular survivin mRNA

Benefiting from the outstanding signal amplification effect and the admirable construction flexibility, the currently proposed DNA motors (particularly DNA walkers) based biosensing concepts have provided a forceful fluorescence imaging tool for intracellular detection. Even so, this promising sensing means is not only subject to poor controllability and prone to produce false signals but also requires exogenous powering forces owing to the common employment of DNAzyme. In response to these challenges, we are herein motivated to present some meaningful solving strategies. For one thing, the surfaces of gold nanoparticles are conducted with a photo-gated walking behavior by introducing a photocleave mode, under which the light-switchable DNA walkers are capable of being selectively activated via an external ultraviolet source to faultlessly prevent the sensing frame from being pre-initiated during cellular uptake and intracellular delivery. For another, the intracellular biothiols are consumed by MnO2 nanosheets to effectively avoid the competitions to Au–S bonds to eliminate potential false outputs and also self-supply sufficient cofactors (Mn2+) to actualize a self-powered operation pattern as well as facilitate the endocytosis process. Following these breakthroughs, a favorable analysis performance towards a model tumor biomarker (survivin mRNA) is endowed with the newly raised biosensor, whose sensitivity is low to pM level with a sound specificity for identifying single base mismatching. Moreover, the significantly improved autonomous three-dimensional DNA walkers can be used to determine and dynamically trace the targets in live cancer cells with an exceptional precise and efficient manner, commendably impelling the sensing ability of DNA motors in biological specimens.

Ultra-sensitive detection of Cr2O72− in farmland achieved by an electrosynthesized fluorescent poly(Fmoc-succinimide)

Hexavalent chromium (Cr(VI)) pollution is related to human health, and the detection of its concentration has a decisive effect on pollution treatment. In this work, we designed a Cr2O72− sensor P(Fmoc-Osu) and applied it in farmland. In the boron trifluoride diethyl etherate solution, the monomer Fmoc-Osu was electropolymerized and obtained its conjugated polymer P(Fmoc-Osu). The introduction of succinimide and carbonic ester group in side chain enabled it to show high selectivity to Cr2O72− in comparison to anions and twenty amino acids. Furthermore, by taking advantage of the signal amplification effect of a conjugated polymer, P(Fmoc-Osu) could analyze Cr2O72− in trace amounts and the limit of detection was 8.3 fM. For agricultural application, we employed it in farmland water, soil, Chinese cabbage, carrot and the results showed a satisfactory result. All these results showed that P(Fmoc-Osu) is an efficient tool to detect Cr2O72− in practical application.

Minimalist Design for a Hand-Held SARS-Cov-2 Sensor: Peptide-Induced Covalent Assembly of Hydrogel Enabling Facile Fiber-Optic Detection of a Virus Marker Protein.

The rampaging COVID-19 needs bioassaying methods of low cost and high robustness for those living in the poorly developed regions. Here, we propose such a method that does not need expensive and complicated equipment. Only a set of hand-held small devices is sufficient. A section along an optic fiber cable is stripped, so that laser light travelling through it will leak outside, while biosensing process taken place on this stripped section can form a new cladding layer of hydrogel, restoring the laser output of the fiber. A short peptide probe immobilized on the stripped section of the fiber can covalently capture a biomarker protein of SARS-Cov-2 from the serum sample. Through the cross-linking of the target protein with the interfering proteins in the serum sample, a hydrogel is covalently immobilized around the stripped section, highly resistant to detergent rinsing that is indispensable for removing nonspecific interference from the clinical sample. Using this "covalent biosensing" strategy, only one peptide probe is sufficient to simultaneously achieve ultrahigh affinity toward the biomarker protein of SARS-Cov-2 and effective signal amplification.

Visual and quantitative detection of E. coli O157:H7 by coupling immunomagnetic separation and quantum dot-based paper strip.

Simple and visual quantitative detection of foodborne pathogens can effectively reduce the outbreaks of foodborne diseases. Herein, we developed a simple and sensitive quantum dot (QD)-based paper device for visual and quantitative detection of Escherichia coli (E. coli) O157:H7 based on immunomagnetic separation and nanoparticle dissolution-triggered signal amplification. In this study, E. coli O157:H7 was magnetically separated and labeled with silver nanoparticles (AgNPs), and the AgNP labels can be converted into millions of Ag ions, which subsequently quench the fluorescence of QDs in the paper strip, which along with the readout can be visualized and quantified by the change in length of fluorescent quenched band. Owing to the high capture efficiency and effective signal amplification, as low as 500cfumL-1 of E. coli O157:H7 could be easily detected by naked eyes. Furthermore, this novel platform was successfully applied to detect E. coli O157:H7 in spiked milk samples with good accuracy, indicating its potential in the detection of foodborne pathogens in real samples.

Ultrasensitive Microfluidic Paper-Based Electrochemical/Visual Analytical Device via Signal Amplification of Pd@Hollow Zn/Co Core-Shell ZIF67/ZIF8 Nanoparticles for Prostate-Specific Antigen Detection.

An effective signal amplification strategy is essential to enhance the analytical performance of microfluidic paper-based analytical devices (μPADs) for tracing biomarkers. Here, a simple but efficient approach with superior electrocatalytic performance of Pd@hollow Zn/Co core-shell ZIF67/ZIF8 nanoparticles for regulating the efficacious signal amplification process was utilized to realize the detection of prostate-specific antigen (PSA). By rationally designing the core-shell structure of ZIF67/ZIF8 with hollow characteristics on the nanoscale and introducing the noble metal element Pd into the cavity, the diffusion limitation and porous confinement reduction of the obtained nanomaterials with uniform morphology and satisfactory chemical stability could be realized, which endowed it with better catalytic performance than solid metal-organic frameworks (MOFs) and ensured effective signal amplification of H2O2 reduction for achieving enhanced electrochemical signals. Moreover, with the assistance of signal probes, the remaining H2O2 could flow to the color area to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine to form a colored product by changing the spatial configuration of the μPAD, thus realizing the visual detection of PSA. On the basis of this novel analytical device, dual-mode ultrasensitive detection of PSA could be achieved with a lower limit of detection of 0.78 pg/mL (S/N = 3) and a wider linear range from 5 pg/mL to 50 ng/mL. This work provided the opportunity of introducing the noble metal element Pd into the cavity of the MOF hollow structure to improve its electrocatalytic efficiency and construct a high-performance μPAD for clinical detection of other biomarkers.

Periodically programmed building and collapse of DNA networks enables an ultrahigh signal amplification effect for ultrasensitive nucleic acids analysis

Analysisof molecular species is needed for applications in diagnosis of infections and genetic diseases. Herein, we demonstrate a target DNA-responsive ultrahigh fluorescence signal-on DNA amplification system via periodically programmed building and collapse of DNA networks. In this system, a pair of oligonucleotides of padlock probe (PP) and palindromic hairpin probe (PHP) are utilized. The presence of target DNA firstly hybridizes with PP, allowing the occurrence of rolling circle amplification (RCA) to produce RCA products with tandem repeats in abundance to bind and unfold numbers of PHPs. The conformational change of PHPs enables the building of DNA networks via the intermolecular palindrome pairing, but then makes the DNA networks collapsed via the palindrome-induced strand displacement polymerization. The displaced RCA products are dynamically reused to undergo periodically programmed multiple rounds of DNA network building and collapse. Depend on the bidirectional DNA assembly and disassembly, a strikingly amplified fluorescence can be collected to ultrasensitive and specific detection of target DNA. The practicability has been demonstrated by evaluating target-spiked human serum, saliva, and urine samples with acceptable recoveries and reproducibility. Therefore, this newly explored method opens a promising avenue for the detection of nucleic acids with low abundance in biochemical analysis and diseases diagnosis.