Amplification Strategy Research Articles

Integrated triple signal amplification strategy for ultrasensitive electrochemical detection of gastric cancer-related microRNA utilizing MoS2-based nanozyme, hybridization chain reaction, and horseradish peroxidase

Early diagnosis and treatment of gastric cancer (GC) play a vital role in improving efficacy, reducing mortality and prolonging patients’ lives. Given the importance of early detection of gastric cancer, an electrochemical biosensor was developed for the ultrasensitive detection of miR-19b-3p by integrating MoS2-based nanozymes, hybridization chain reaction (HCR) with enzyme catalyzed reaction. The as-prepared MoS2-based nanocomposites were used as substrate materials to construct nanoprobes, which can simultaneously load probe DNA and HCR initiator for signal amplification. Moreover, the MoS2-based nanocomposites are also employed as nanozymes to amplify electrochemical response. The presence of miR-19b-3p induced the assembly of MoS2-based nanoprobes on the electrode surface, which can activate in-situ HCR reaction to load a large number of horseradish peroxidase (HRP) for signal amplification. Coupling with the co-catalytic ability of HRP and MoS2-based nanozymes, the designed electrochemical biosensor can detect as low as 0.7 aM miR-19b-3p. More importantly, this biosensor can efficiently analyze miR-19b-3p in clinical samples from healthy people and gastric cancer patients due to its excellent sensitivity and selectivity, suggesting that this biosensor has a potential application in early diagnosis of disease.

Intelligent nanocatalyst mediated lysosomal ablation pathway to coordinate the amplification of tumor treatment

The production of reactive oxygen species (ROS) is susceptible to external excitation or insufficient supply of related participants (e.g., hydrogen peroxide (H2O2) and sensitizer), liming ROS-driven tumor treatment. Additionally, the lysosomal retention effect severely hinders the utilization of ROS-based nanosystems and severely restricted the therapeutic effect of tumors. Therefore, first reported herein an intelligent nanocatalyst, TCPP-Cu@MnOx ((MnII)1(MnIII)2.1(MnIV)2.6O9.35), and proposed a programmed ROS amplification strategy to treat tumors. Initially, the acidity-unlocked nanocatalyst was voluntarily triggered to generate abundant singlet oxygen (1O2) to mediate acid lysosomal ablation to assist nanocatalyst escape and partially induce lysosomal death, a stage known as lysosome-driven therapy. More unexpectedly, the high-yielding production of 1O2 in acid condition (pH 5.0) was showed compared to neutral media (pH 7.4), with a difference of about 204 times between the two. Subsequently, the escaping nanocatalyst further activated H2O2-mediated 1O2 and hydroxyl radical (•OH) generation and glutathione (GSH) consumption for further accentuation tumor therapy efficiency, which is based on the Fenton-like reaction and Russell reaction mechanisms. Therefore, in this system, a program-activatable TCPP-Cu@MnOx nanocatalyst, was proposed to efficiently destruct organelle-lysosome via1O2 inducing, and stimulated H2O2 conversion into highly toxic 1O2 and •OH in cytoplasm, constituting an attractive method to overcome limitations of current ROS treatment.

Highly sensitive β-amyloid1–42 oligomer aptamer-based assay via dual cyclic amplifications of three-way catalytic hairpin assembly and palindrome polymerase reaction

β-amyloid1–42 oligomer (AβO1–42), a neurotoxic protein, is considered the key factor in the progression of Alzheimer's disease (AD). And, high sensitivity detection of AβO1–42 is of considerable importance for the early diagnosis and deterioration prevention of AD. We present here an ultrasensitive fluorescent AβO1–42 detection approach based on aptamer recognition probes and a new dual cyclic signal amplification strategy by combining three-way catalytic hairpin assembly (3W-CHA) with palindromic sequence-based polymerase amplification (PBPA). Once the aptamers bind to target AβO1–42 molecules, ssDNA sequences are released to initiate 3W-CHA between three hairpins to unfold the signal hairpins and to form Y-shaped DNA structures (Y-DNAs). Subsequently, the assistance ssDNAs bind the Y-DNAs to expose the palindromic sequences to yield Y-DNA dimers, which lead to cyclic PBPA reaction with presence of polymerase to further unfold more signal hairpins for yielding considerably magnified fluorescence recovery, enabling sensitive assay of AβO1–42 down to 2.4 pM within a dynamic range of 5 pM∼50 nM. Furthermore, this strategy has been validated for interrogation of low AβO1–42 levels in artificial cerebrospinal fluid (CSF), suggesting its promising potential for application in early diagnosis and prevention of AD.

Synergistic signal amplification for HER2 biosensing using tetrahedral DNA nanostructures and 2D transition metal carbon/nitride@Au composites via ARGET ATRP

The rising incidence of breast cancer underscores the critical need for accurate and early detection methods, particularly for key biomarkers like human epidermal growth factor receptor 2 (HER2). Here, a synergistic signal amplification electrochemical biosensor was developed for HER2 detection that utilizes tetrahedral DNA nanostructures (TDN), 2D transition metal carbon/nitride@Au (MXene@Au) composites, and activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP). Specifically, TDN serves as the substrate, with the aptamer anchored by the MXene@Au composite support matrix. This design facilitates the generation of electrochemical currents through the ARGET ATRP signal amplification strategy. The robustness and high affinity of TDN minimize surface effects on the electrode and enhance target binding. Additionally, the high conductivity of the MXene@Au composites improves the detection sensitivity of the biosensor. The use of the ARGET ATRP strategy further enhances detection sensitivity. The biosensor exhibits a wide detection range from 10 pg·mL−1 to 10 µg·mL−1 with a remarkable detection limit of 0.39 pg·mL−1, verified under optimal experimental conditions. The designed biosensor offers a significant advancement in the field of oncological diagnostics, providing a highly sensitive, low-cost, and simple method for early HER2 detection, thereby potentially improving treatment outcomes for breast cancer patients.

Advanced design of target-driven self-powered sensor assisted by cascade catalytic strategy

In this work, a self-powered microsensor platform based on enzyme biofuel cells (EBFCs) was developed for intelligent monitoring of disease markers miRNA-451. The cascade catalysis system constructed by using the strategy of enzyme-like ZIF-8 nanocapsule incorporation with biological enzymes, which could simultaneously take into account the specificity of biological enzymes and the high activity of nano-enzymes, significantly promoted the electron transfer between glucose and the bio-anode surface, and improved the sensitivity and stability of the sensing system. Meanwhile, the target-triggered hybridisation chain reaction (HCR) amplification strategy to achieve exponential signal amplification based on accurate recognition, and jointly improve the detection sensitivity. As expected, the micro-sensor platform has a wide linear range of 0.5-1.0 fmol/L with a low limit of detection (LOD) of 0.13 fmol/L (S/N=3) and exhibits excellent selectivity, reproducibility and stability in interference assays under optimal detection conditions. The designed self-powered system is simple to construct, easy to transport and the data transmission mode is intelligent and controllable, which is expected to be used in basic biochemical research, clinical diagnosis and environmental monitoring.

Enhancement of single-photon level signal detection based on phase sensitive amplification strategy

Abstract We investigate the ability of phase-sensitive amplification (PSA) to noiselessly amplify the preferred quadrature components of single-photonsignals that are limited by phase fluctuations relative to the pump. We presenta PSA enhancement strategy that is more realistic for possible experimentalrealization and is expected to significantly improve the detection of weaksignals at the single-photon level and obtain more useful information. Our studyshows that with a large PSA gain, proper transmissivity allows both direct and
balanced homodyne detections to operate optimally simultaneously, and effectively improves the noise figure degradation owing to the internal losses and non-ideal detection efficiency.

Enzyme-accelerated catalytic DNA circuits enable rapid and one-pot detection of bacterial pathogens

Catalytic DNA circuits, serving as signal amplification strategies, can enable simple and accurate detection of pathogenic bacteria in complex matrices but suffer from low reaction rates and depths. Herein, we design an enzyme-accelerated catalytic hairpin assembly (EACHA) in which duplex DNA products are converted into hairpin reactants to continue participating in the next circuit reaction with the assistance of RNase H. Profiting from the high recyclability of the reactants, EACHA exhibits an approximately 37.6-fold enhancement in the rate constant and a two-order-of-magnitude improvement in sensitivity compared to conventional catalytic hairpin assembly (CHA). By integrating an allosteric probe with EACHA, a one-pot method is developed for rapid and direct detection of S. enterica Enteritidis (S. Enteritidis). This method is capable of detecting 15 CFU mL−1 of S. Enteritidis within 20 min, which is superior to that of real-time PCR. By testing 60 milk samples, we demonstrate this method's high accuracy in discriminating contaminated samples, with an area under the curve (AUC) of 0.997. Moreover, this method can be employed to accurately diagnose early-stage infected mice, with an AUC of 1.00 for feces samples and 0.986 for serum samples. Therefore, this study offers a simple and feasible method for identifying pathogens in complex matrices.

CRISPR/Cas13a-mediated triple signal amplification strategy for viral RNA detection

CRISPR/Cas13a system has been combined with different isothermal amplification strategies for RNA analysis. Nevertheless, expensive enzymes are usually indispensable in these isothermal amplification methods, which increases the detection cost. In this study, a CRISPR/Cas13a-mediated triple signal amplification strategy has been designed for selective, sensitive and inexpensive detection of viral RNA. SARS-CoV-2 S RNA activated CRISPR/Cas13a to cut hairpin DNA, triggering catalytic hairpin assembly (CHA) and subsequent hybridization chain reaction (HCR) to form a long-stranded Y-type DNA. G-quadruplexes were distributed on the branch of the long-stranded DNA and bound with N-methyl mesoporphyrin IX (NMM) to enhance the fluorescence signal. SARS-CoV-2 S RNA was specifically detected with a limit of detection (LOD) of 285 fM. Notably, no additional expensive enzymes were required in the entire detection process. The CRISPR/Cas13a-mediated triple signal amplification strategy holds great potential for specific and sensitive viral RNA detection with low cost.

Aptamer-Mediated Electrochemical Detection of SARS-CoV-2 Nucleocapsid Protein in Saliva.

Gold standard detection of SARS-CoV-2 by reverse transcription quantitative PCR (RT-qPCR) can achieve ultrasensitive viral detection down to a few RNA copies per sample. Yet, the lengthy detection and labor-intensive protocol limit its effectiveness in community screening. In view of this, a structural switching electrochemical aptamer-based biosensor (E-AB) targeting the SARS-CoV-2 nucleocapsid (N) protein was developed. Four N protein-targeting aptamers were characterized on an electrochemical cell configuration using square wave voltammetry (SWV). The sensor was investigated in an artificial saliva matrix optimizing the aptamer anchoring orientation, SWV interrogation frequency, and target incubation time. Rapid detection of the N protein was achieved within 5 min at a low nanomolar limit of detection (LOD) with high specificity. Specific N protein detection was also achieved in simulated positive saliva samples, demonstrating its feasibility for saliva-based rapid diagnosis. Further research will incorporate novel signal amplification strategies to improve sensitivity for early diagnosis.

One-step cascade amplification system based on entropy-driven catalysis and DNAzyme triggered DNA walker for label-free detection of acetamiprid

Herein, an electrochemical aptasensor for highly sensitive detection of acetamiprid (ACE) was constructed based on a one-step cascade amplification strategy. This innovative strategy integrated DNA walker containing DNAzyme sequence into entropy-driven catalysis (EDC) system. The trigger strand was released by aptamer-specific binding to ACE, initiating the EDC amplification circuit and delivering DNA walker strands. The dangling DNA walker continuously bound and cleaved hairpin substrate to form G-quadruplex fragments with the assistance of Mg2+. The G-quadruplex fragments folded and captured hemin to form multitudinous G-quadruplex/hemin complexes in the presence of K+, generating significantly enhanced current, enabling enzyme-free, label-free and highly sensitive detection of ACE, with a linear detection range of 100 fM to 50 nM and a detection limit of 68.36 fM (S/N = 3). The constructed aptasensor achieved the reliable detection of ACE in vegetable soil and cucumber samples, demonstrating its potential application prospects in environmental protection and food supervision.

Cu2O-Mediated Heterojunction Conversion from Dual Type II to Dual Z-Scheme: Its Application in Photoelectric-Colorimetric Dual-Mode Detection of Fat Mass and Obesity-Associated (FTO) Protein.

Although the construction of heterojunction has been used in photoelectrochemical (PEC) biosensors, their potential for tunable optical properties has not been deeply explored. Based on the fact that a type-II heterojunction and Z-scheme heterojunction have the same energy band structure, effective alteration of the electron transfer pathway has been achieved by introducing unique photoactive materials into the system and exploiting the interactions between the photomaterials. Based on this, we reported a novel polarity-switchable dual-mode sensor for fat mass and obesity-associated (FTO) protein analysis. Specifically, the MgIn2S4/Bi2MoO6/Bi2S3 dual type-II heterojunction was used as the sensing interface in concert with the rolling circle amplification, CRISPR/Cas12, and terminal DNA transfer enzyme multiamplification strategies, and finally, Cu2O was captured at the sensing interface. Due to the matched energy band, the introduction of Cu2O effectively changed the electron transfer pathway and realized the conversion from a dual type-II heterojunction to a dual Z-scheme heterojunction. It caused the switch of the photocurrent from the anode to the cathode. The developed PEC method showed high sensitivity and selectivity for FTO protein detection in the range of 0.0005-500 μg/L. In addition, based on the peroxidase-like activity of Cu2O to catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine by H2O2, the electrode system also achieved the colorimetric detection of FTO protein using the naked eye with the change of the color of the detection solution from colorless to blue. The detection range was from 0.05 to 500 μg/L. This work developed a photoelectrochemical-colorimetric biosensing platform with consciously designed semiconductor structures, revealing the potential of semiconductor-structured transformations in future sensing fields.

Highly sensitive detection of RNase H via a novel DNA/RNA heteroduplex combining isothermal exponential amplification strategy

Ribonuclease H (RNase H) plays a crucial role in a variety of cellular processes and is emerging as an essential therapeutic target for many diseases. Various methods have been constructed to assay RNase H activity, but these methods are either time-consuming or have poor sensitivity. In this study, we developed a novel strategy that combined a specially designed DNA/RNA chimeric substrate with exponential amplification reaction (EXPAR) for rapid and sensitive detection of RNase H activity. In the presence of RNase H, the RNA strand of the DNA/RNA heteroduplex was specifically degraded, forming a 3′-hydroxyl group in the locking primer for recognition and extension by DNA polymerase. This ultimately triggered EXPAR and produced a large number of single-stranded DNA, which was monitored in real-time with fluorescent dye. Under optimized conditions, the proposed strategy can detect as low as 6 × 10−10 U/μL of RNase H, which was at least 1000 times more sensitive than several reported methods. Furthermore, we demonstrated the usage of this method for RNase H inhibitor analysis and practical application in complex biological samples, including serum and tumor cell extracts. Therefore, these results suggested that the developed method is a promising tool for highly sensitive detection of RNase H and RNase H-associated disease diagnosis.

Self-reinforced D-MOF and autocatalytic AuPd@SnS2 dual-wavelength electrochemiluminescence biosensor for detection of ctDNA and CEA

Trace detection of circulating tumor DNA (ctDNA) and CEA is of great significance for early prevention and diagnosis of cancer. In this work, the well electrochemiluminescence (ECL) of the dual-ligand metal-organic frameworks (D-MOFs) and AuPd@SnS2 QDs nuclear-satellite conjugate was newly explored, and a dual-wavelength biosensing platform based on ECL energy transfer between D-MOFs and AuPd NPs coupled with catalytic effect of AuPd NPs on ECL of SnS2 QDs was developed for detection of trace ctDNA and CEA. Here, thanks to the ordered heterogeneous arrangement and network charge transfer of the luminescent ligand, the prepared D-MOF exhibits highly enhanced ECL without exogenous co-reactants. Target ctDNA was exponentially amplified using an entropy-driven DNA cycle amplification strategy to obtain bandage DNA, which triggered the DNA walker to connect the AuPd@SnS2 core-satellite probe to the electrode. Therefore, the ECL ratio of D-MOF and sensitized SnS2 at 535 nm and 650 nm was used to achieve ultrasensitive detection of trace ctDNA. Moreover, the specific recognition of target CEA to T1 DNA induced changes in dual-wavelength signals to achieve further detection of CEA, and ensure the accuracy of cancer analysis. The work opened up two novel ECL materials and developed a novel ECL biosensor for rapid detection of double tumor markers, providing a new way for early prevention and diagnosis of actual cancer.

Impact of Different Hearing Aid Amplification Strategies on Speech Recognition in Hearing-Impaired Urdu Speaking Children: A Comparative Study

Impact of Different Hearing Aid Amplification Strategies on Speech Recognition in Hearing-Impaired Urdu Speaking Children: A Comparative Study

Low-cost signal enhanced colorimetric and SERS dual-mode paper sensor for rapid and ultrasensitive screening of mercury ions in tea

Mercury ions (Hg2+) are highly toxic heavy metals that are commonly found in natural environments. Owning to their non-biodegradability and accumulation in the food chain, the precise detection of trace amounts of Hg2+ is essential for preventing chronic accumulation and ensuring food safety. In this study, we present a dual-mode paper sensor for simultaneous colorimetric and Surface-Enhanced Raman Spectroscopy (SERS) detection of Hg2+ in tea, achieving ultrasensitive, rapid, and on-site screening. 4-Mercaptopyridine (4-MPY) was effectively chemisorbed onto the gold nanoparticles (AuNPs), acting as a signal probe for colorimetric methods. Moreover, it can produce plasmonic hot spots for SERS by interacting with the pyridine ring. To enhance the signal intensity of both colorimetry and SERS, a silver shell is in-situ grown on the surface of AuNPs captured on the paper sensor by reduction of Ag+, achieving signal amplification. The visual limit of detection (LOD) for the colorimetric biosensor is 2.5 pM, while the LOD of SERS is 0.48 pM with this dual-mode paper sensor. The sensitivity of both the colorimetric method and SERS was improved by approximately 200 and 500 times, respectively, with the designed signal amplification strategy. The system allows for multiple parallel screening of the same sample, ensuring accurate results without any false-positive or false-negative. This study provides a valuable platform for the accurate detection of various other heavy metal ions and provides effective strategies for improving the performance of colorimetric methods.

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.

Simultaneous detection of CaMV35S and NOS using fluorescence sensors with dual-emission silver nanoclusters and catalytic hairpin amplification strategy.

Adual-emission fluorescent biosensing method was developed for simultaneous determinationof CaMV35S and NOS in genetically modified(GM) plants. Two designed hairpin DNA (H1, H2) sequences were used as templates to synthesize H1-AgNCs (λex = 570nm, λem = 625nm) and H2-AgNCs (λex = 470nm, λem = 555nm). By using H1-AgNCs and H2-AgNCs as dual-signal tags, combined with signal amplification strategy of magnetic separation to reduce background signal and an enzyme-free catalytic hairpin assembly (CHA) signal amplification strategy, a novel multi-target fluorescent biosensor was fabricated to detect multiple targets based on FRET between signal tags (donors) and magnetic Fe3O4 modified graphene oxide (Fe3O4@GO, acceptors). In the presence of the target NOS and CaMV35S, the hairpin structures of H1 and H2 can be opened respectively, and the exposed sequences will hybridize with the G-rich hairpin sequences HP1 and HP2 respectively, displacing the target sequences to participate in the next round of CHA cycle. Meanwhile, H1-HP1 and H2-HP2 double-stranded DNA sequences (dsDNA) were formed, resulting in the desorption of dsDNA from the surface of Fe3O4@GO due to weak π-π interaction between dsDNA and Fe3O4@GO and leading to the fluorescence recovery of AgNCs. Under optimal conditions, the linear ranges of this fluorescence sensor were 5 ~ 300nmol L-1 for NOS and 5 ~ 200nmol L-1 CaMV35S, and the LODs were 0.14nmol L-1 and 0.18nmol L-1, respectively. In addition, the fluorescence sensor has good selectivity for the detection of NOS and CaMV35S in GM soybean samples, showing the potential applications in GM screening.

Advanced DNA Amplification for Efficient Data Storage.

DNA amplification technologies have significantly advanced biotechnology, particularly in DNA storage. However, adaptation of these technologies to DNA storage poses substantial challenges. Key bottlenecks include achieving high throughput to manage large data sets, ensuring rapid and efficient DNA amplification, and minimizing bias to maintain data fidelity. This perspective begins with an overview of natural and artificial amplification strategies, such as polymerase chain reaction and isothermal amplification, highlighting their respective advantages and limitations. It then explores the prospective applications of these techniques in DNA storage, emphasizing the need to optimize protocols for scalability and robustness in handling diverse digital data. Concurrently, we identify promising avenues, including advancements in enzymatic processes and novel amplification methodologies, poised to mitigate existing constraints and propel the field forward. Ultimately, we provide insights into how to utilize advanced DNA amplification strategies poised to revolutionize the efficiency and feasibility of data storage, ushering in enhanced approaches to data retrieval in the digital age.

Mg2+- dependent six-jawed trap-mediated SERS biosensor for ratiometric detection of microRNA with use of the highly ordered Au NCs as substrate

Herein, a ratiometric surface-enhanced Raman scattering (SERS) biosensor was constructed with use of the highly ordered gold nanocubes (Au NCs) interface created by the liquid-phase assembly as substrate, and a novel Mg2+-dependent six-Jawed trap (SJT) cyclical amplification strategy for sensitive detection of microRNA 221 (miRNA-221). Impressively, in contrast to conventional gold nanoparticles (Au NPs), Au NCs with more sharp corners and edge characteristics can significantly improve the Raman signals of the Raman molecules, while the highly ordered Au NCs prepared by liquid-phase assembly can also increase the stability and repeatability of miRNA-221 detection. In addition, the newly constructed SJT with more Mg2+ shear sites were utilized to efficiently release plentiful output DNA with the Raman molecule methylene blue (MB) for greatly enhancing the sensitivity of the SERS biosensor. Ultrasensitive ratiometric SERS detection of miRNA-221 was realized by capturing the output DNA containing MB to generate ratiometric Raman signals at the sensing interface modified with Cyanine 5 (Cy5) Raman molecules. The SERS biosensor analyzed for miRNA-221 from 1 fM - 10 nM with an identification limit of 0.66 fM, and the approach is also effective for detecting miRNA-221 in cell lysates. Notably, the SJT signal amplification strategy utilized in this study makes it easier to create extremely sensitive biosensors and opens up new avenues for early detection of hepatocellular carcinoma cells (HCC).

CRISPR/Cas12a powered sandwich aptamer assay coupled with effective DNA self-assembly amplification for sensitive detection of immunoglobulin E

Sensitive and accurate detection of protein biomarkers is of significance for disease diagnosis. Recently, CRISPR/Cas system has emerged as a powerful signal amplifier in the field of sensing technology. We report a CRISPR/Cas12a powered sandwich aptamer assay (CAPSA) for protein analysis coupled with effective DNA self-assembly amplifications. The target protein is sandwiched between the capture antibody on the microplate and the aptamer detection probe. The aptamer is engineered with a specific initiator to trigger streptavidin (SA)-biotin-based dendritic amplification reaction, yielding a DNA polymer with multiple repeats of active DNA (acDNA) as Cas12a recognition sites. With the addition of Cas12a reporter system, the acDNA activates Cas12a to cleave DNA reporters, producing amplified fluorescence signals. Particularly, we also describe an innovative one-step DNA self-assembly amplification strategy to meet the demands for fast point-of-care testing, which can be directly integrated into the CAPSA without additional operation steps. The proposed methods were demonstrated by detecting immunoglobulin E (IgE), which plays a vital role in allergic reactions. Our proposed one-step DNA self-assembly amplified CAPSA enabled us to detect IgE as low as 0.6 pM, exhibiting high selectivity and satisfactory performance in complex sample analysis. Collectively, this method combines the efficient signal amplification of DNA self-assembly with the high trans-cleavage activity of Cas12a, offering a simple, universal, and double-amplified approach for sensitive analysis of protein biomarkers.