Chemosensors
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2022
·Vol. 10
·期 7
·276
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IF 4.229
Kunhan Nie,Yongjian Jiang,Na Wang,Yajun Wang,Di Li,Lei Zhan,Chengzhi Huang,Chunmei Li
The abnormal expression of miRNA is closely related to the occurrence of pancreatic cancer. Herein, a programmable DNAzyme amplifier for the universal detection of pancreatic cancer-related miRNAs was proposed based on its programmability through the rational design of sequences. The fluorescence signal recovery of the DNAzyme amplifier showed a good linear relationship with the concentration of miR-10b in the range of 10–60 nM, with a detection limit of 893 pM. At the same time, this method displayed a high selectivity for miR-10b, with a remarkable discrimination of a single nucleotide difference. Furthermore, this method was also successfully used to detect miR-21 in the range of 10–60 nM based on the programmability of the DNA amplifier, exhibiting the universal application feasibility of this design. Overall, the proposed programmable DNAzyme cycle amplifier strategy shows promising potential for the simple, rapid, and universal detection of pancreatic cancer-related miRNAs, which is significant for improving the accuracy of pancreatic cancer diagnosis.Keywords:DNAzyme amplifier;signal amplification;miRNA detection;pancreatic cancer
研究领域:
分子生物学
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医学诊断
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生物化学
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DOI
高影响因子
TALANTA
·
2025
·
IF 6.1
Tao Meng,Qi Kang,Jiashuo Xu,Shuang Zhao,Tianqi Liu,Dianming Zhou,Xiaoqun Gong,Jianyu Zhang
The CRISPR/Cas12a system has become a powerful tool in biosensing because of its specific target recognition ability and highly efficient trans -cleavage activity. However, a problem faced by the CRISPR/Cas12a system when directly used for trace detection is the linear amplification efficiency of single-cycle digestion. Here, we present a novel hairpin reporter-driven CRISPR/Cas12a (HR-CRISPR) amplification system that establishes a positive feedback loop within the CRISPR/Cas12a platform to finish an exponential and sensitive signal amplification in a one-step reaction. As proof of concept, we applied this strategy to the terminal deoxynucleotidyl transferase (TdT) activity assay without pre-amplification procedure. The polyT strand extended by TdT hybridizes with crRNA, activating Cas12a, which then cleaves the FQ-hairpin reporter. The cleavage products are further elongated by reverse transcriptase using crRNA as a template, reactivating Cas12a and producing exponentially amplified fluorescence signals. This assay offers a simple yet highly sensitive approach for quantifying TdT activity, achieving a low detection limit of 4.55 × 10 −6 U. Moreover, it is applicable for inhibitor screening and monitoring TdT activity in human serum samples.
研究领域:
分子生物学
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生物技术
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医学检测
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2025
·
IF 5.1
Ping Qiu,Longjian Huang,Jiayi Zhang,Wenbo Li,Xudong Xin,Luchang Chen,Qianli Tang,Kai Zhang,Xianjiu Liao,Jihua Wei
We report the development of a novel split aptamer-driven electrochemical biosensor integrating entropy-driven Signal amplification (EDSA) and CRISPR-Cas12a technology for the highly sensitive and specific detection of adenosine deaminase (ADA). ADA is an important biomarker involved in immune regulation and purine metabolism, and its accurate quantification is critical for diagnosing various diseases. In our system, ADA catalyzes the deamination of adenosine to inosine, resulting in the release of Aptamer1 from a split aptamer structure. The liberated Aptamer1 initiates a strand displacement reaction within a Blocker/Bandage/Scaffold complex, triggering a cascade of entropy-driven amplification cycles that generate a DNA duplex capable of activating CRISPR-Cas12a. Activated Cas12a exhibits trans -cleavage activity, specifically cleaving a ferrocene-labeled single-stranded DNA (ssDNA) probe immobilized on a gold electrode, leading to a measurable decrease in electrochemical signal. The biosensor operates under isothermal and enzyme-free conditions, providing simplicity, rapid detection, and low cost. It achieves a low detection limit of 0.688 U·L −1 with a wide linear range from 1 to 20,000 U·L −1 . The system demonstrates excellent specificity against ADA analogues and common interfering proteins and performs reliably in diluted human serum samples. This strategy represents a versatile and modular platform for point-of-care diagnostics. By altering the aptamer sequence, the biosensor can be adapted for the detection of other clinically relevant proteins and enzymes, contributing to advances in personalized medicine.
研究领域:
生物传感技术
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分子诊断
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医学检测
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DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Yuting Xu,Lili Li,Yumeng Yang,Chengzhi Huang,Hongyan Zou
Developing a simple, reliable, and sensitive hepatitis C virus (HCV) genetic sensing platform is of great significance for diagnosing diseases and selecting appropriate antiviral treatments. Herein, a tandem nucleic acid amplification strategy for sensitive detection of HCV genotype 1b (HCV-1b) was developed by stringing the catalytic hairpin assembly (CHA) and the triggered DNAzyme amplifier. The hairpin reactants were initiated by the target to produce lots of triggering double-stranded DNA sequences which can efficiently activate the subsequent blocked DNAzyme. Thereby, the continuous cleavage of substrate was realized, resulting in the fluorescence signal amplification. The DNA-based isothermal CHA-DNAzyme (CDz) sensing platform was successfully applied for sensitive detection of HCV-1b with the limit of detection (84 pM) and showed good selectivity. Moreover, the practical detection of target DNA in the complex biologic matrix indicated that the developing strategy had good potential for early HCV infection diagnosis.
研究领域:
分子生物学
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病毒学
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诊断学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2021
·
IF 6.986
Sha Liu,Siyu Lu,Shihao Sun,Jun Hai,Genping Meng,Baodui Wang
Quantitative detection of cancer cells using portable devices is promising for the development of simple, fast, and point-of-care cancer diagnostic techniques. However, how to further amplify the detection signal to improve the sensitivity and accuracy of detecting cancer cells by portable devices remains a challenge. To solve the problem, we, for the first time, synthesized folic-acid-conjugated Au nanoframes (FA-Au NFs) with amplification of pressure and temperature signals for highly sensitive and accurate detection of cancer cells by portable pressure meters and thermometers. The resulting Au NFs exhibit excellent near-infrared (NIR) photothermal performance and catalase activity, which can promote the decomposition of NH4HCO3 and H2O2 to generate corresponding gases (CO2, NH3, and O2), thereby synergistically amplifying pressure signals in a closed reaction vessel. At the same time, Au NFs with excellent peroxidase-like activity can catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to produce TMB oxide (oxTMB) with a strong photothermal effect, thereby cooperating with Au NFs to amplify the photothermal signal. In the presence of cancer cells with overexpressing folate receptors (FRs), the molecular recognition signals between FA and FR can be converted into amplified pressure and temperature signals, which can be easily read by portable pressure meters and thermometers, respectively. The detection limits for cancer cells using pressure meters and thermometers are 6 and 5 cells/mL, respectively, which are better than other reported methods. Moreover, such Au NFs can improve tumor hypoxia by catalyzing the decomposition of H2O2 to produce O2 and perform photothermal therapy of cancer. Together, our work provides new insight into the application of Au NFs to develop a dual-signal sensing platform with amplification of pressure and temperature signals for portable and ultrasensitive detection of cancer cells as well as personalized cancer therapy.
研究领域:
生物医学工程
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肿瘤学
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纳米医学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry C
·
2020
·Vol. 8
·期 25
·8515-8520
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IF 7.059
Qian Xiong,Jinlong Yang,Huaiyi Ding,Juan Du,Xiaosheng Tang,Tongchao Shi,Zhengzheng Liu,Daofu Wu,Hao Lin,Yuxin Leng
Recently, solution-processed conventional I–III–VI2 quantum dots (QDs) demonstrated a potential optical amplification application based on superior optical properties. Nonetheless, exploiting the potential of this type of material as a gain medium for optical amplification devices is still scarce, rendering application prospect restricted. Herein, for the first time, we report the amplified spontaneous emission (ASE) action using AgInS2 (AIS) QDs as the gain medium. ASE with an ultra-low threshold (31.58 μW cm−2) was demonstrated by combining highly reflective distributed Bragg reflectors (DBR) with AIS QDs. Furthermore, the relationship between the number of layers of the DBR mirrors and the ASE performance is systematically discussed, indicating that the seven-layer DBR is a desirable device structure with the best ASE from AIS QDs. These consequences unambiguously uncover the significant feasibility of the traditional AIS QDs as a photonic resource, simultaneously demonstrating the huge potential and promising applications of as-designed devices for optical amplification.
研究领域:
材料科学
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光子学
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纳米技术
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2019
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IF 9.518
Zhen Zhang,Mengting Zhu,Zichao Chen,Xiaomeng Wang,Guang Chen,Shusheng Zhang
We herein report the first attempt to engineer a coaxial-sensing 3D amplifier able to achieve dynamic self-assembly in response to a mutated-ctDNA target. A bio-nanofiber is firstly manufactured via an ingenious double-channel electrostatic spinning and DNA rolling circle replication (RCR) technology, which offered an ideal scaffold for assembly of 3D amplifier activated by target recognition. The coaxial-controllable signal amplifier presented several advantages. (1) Given its “coaxial sensing effect”, the proposed bio-amplifier played the coaxial transduction for signal enrichment to vastly increase sensitivity, capable of discriminating a single-base mismatched sequence from the perfectly complementary one, using ctDNA-134A as a model analyte . (2) Due to “covalent bridges lock effect” in an identifying chip with locked nucleic acid beacons, this 3D amplifier expressed high specificity and biostability toward seven different mutated-ctDNAs. (3) Profiting from special configuration of bioactive nanofibers and DNA replication programming, this catalytic bio-amplifier possessed “signal enrichment effect”, which enhanced dynamic range toward ctDNA-134A detection and hybridized without any external indicators . This innovative bio-amplifier has a detection limit of 5.1 aM for ctDNA-134A with superior specificity, excellent sensitivity, and good performance. This pioneered method was further applied for broadly differentiate cells and evaluate changes in the expression levels of intracellular mutated-ctDNAs.
研究领域:
生物医学工程
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分子诊断学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2017
·Vol. 41
·期 15
·7613-7620
·
IF 3.269
Qianqian Pei,Yu Wang,Su Liu,Yifei Qin,Xueqi Leng,Xuejun Cui,Jiadong Huang
In this study, a novel electrochemical biosensor based on exonuclease III (Exo III)-aided autonomous cascade signal amplification for ultrasensitive and highly specific detection of Salmonella typhimurium (S. typhimurium) has been reported. Herein, two duplex DNA probes were ingeniously designed. An Ap–Pr duplex probe was constructed as a recognition element by hybridizing an S. typhimurium aptamer (Ap) with a primer (Pr). A P–S duplex probe has been designed to contain a protruding DNA fragment at 5′-termini that has an identical sequence with Pr (the secondary Pr), a complementary duplex DNA that is used for blocking a signal-triggered probe (S), and an overhung nucleic acid segment at 3′-end that can anneal with Pr and form double-stranded DNA with a blunt 3′-terminus, thus initiating Exo III-aided amplification reaction. Upon sensing of the target S. typhimurium, the specific recognition of S. typhimurium and aptamer releases Pr, which anneals with the P–S duplex and triggers the Exo III cleavage reaction, accompanied by release of Pr, S, and secondary Pr. After this, the released Pr and secondary Pr anneal with other P–S duplex, which in turn triggers the Exo III-assisted cycling cleavage. Thus, large amounts of S are liberated, which hybrids with S* (DNA probe complementary to S); thereby, 5′-methylene blue (MB)-labelled DNA probe (MBP) folds into a hairpin structure with 5′-terminus close to electrode surface for efficient electronic transfer. Moreover, the S–S* hybrid duplex is specifically hydrolyzed by Exo III to liberate S, which accordingly results in the cycling cleavage of S*. Therefore, numerous MBP transform into a hairpin structure with electroactive MB close to the electrode surface; this generates significantly amplified electrochemical signal towards S. typhimurium detection. The present developed strategy exhibited high sensitivity towards S. typhimurium with a detection limit of 2.8 × 10 cfu mL−1. Furthermore, the proposed autonomous cascade signal amplification strategy can be easily extended for detection towards a wide spectrum of analytes including nucleic acid, proteins, and small molecules possessing their corresponding aptamers. Therefore, the developed Exo III-aided autonomous cascade signal amplification strategy might indeed provide a facile and universal tool for bioanalysis and clinical diagnosis.
研究领域:
生物传感技术
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分子诊断
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分析化学
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DOI
高影响因子
Synthetic and Systems Biotechnology
·
2023
·
IF 4.8
Shuting Hu,Guangbao Zhang,Xiaoqiang Jia
In this study, we designed a Cd 2+ whole-cell biosensor with both positive and negative feedback cascade amplifiers in Pseudomonas putida KT2440 (LTCM) based on our previous design with only a negative feedback amplifier (TCM). The results showed that the newly developed biosensor LTCM was greatly improved compared to TCM. Firstly, the linear response range of LTCM was expanded while the maximum linear response range was raised from 0.05 to 0.1 μM. Meanwhile, adding a positive feedback amplifier further increased the fluorescence output signal of LTCM 1.11–2.64 times under the same culture conditions. Moreover, the response time of LTCM for detection of practical samples was reduced from 6 to 4 h. At the same time, LTCM still retained very high sensitivity and specificity, while its lowest detection limit was 0.1 nM Cd 2+ and the specificity was 23.29 (compared to 0.1 nM and 17.55 in TCM, respectively). In summary, the positive and negative feedback cascade amplifiers effectively improved the performance of the biosensor LTCM, resulting in a greater linear response range, higher output signal intensity, and shorter response time than TCM while retaining comparable sensitivity and specificity, indicating better potential for practical applications.
研究领域:
生物技术
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环境科学
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合成生物学
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2021
·
IF 9.229
Senyao Wang,Baoshan He,Ying Liang,Huali Jin,Min Wei,Wenjie Ren,Zhiguang Suo,Jinshui Wang
Herein, a hierarchically porous Zr-MOF-labeled electrochemical aptasensor based on the composite of PtPd@Ni-Co hollow nanoboxes (PtPd@Ni-Co HNBs) and poly (diallyldimethylammonium chloride)-functionalized graphene (PDDA-Gr) was developed for ultrasensitive detection of chloramphenicol (CAP). PtPd@Ni-Co HNBs have excellent conductivity and provide binding sites for aptamers; the functionalized PDDA-Gr improves its dispersibility and conductivity as a substrate material, which can be successfully used to increase the electrode surface area and support more PtPd@Ni-CoHNBs. Besides, hierarchically porous Zr-MOFs (HP-UiO-66) were utilized as signal probes and showed a stronger load capacity for signal molecules than conventional UiO-66. In the presence of CAP, two ingeniously designed Exo III-assisted cyclic amplification strategies further improved the sensitivity of the aptasensor: CAP causes cycle I to release a large amount of trigger DNA (Tr DNA), and then, Tr DNA initiated cycle II, which causes the exposed capture DNA to further bind the signal probes. With these advantages, the constructed aptasensors performed with satisfactory sensitivity in a wide linear range (10 fM–10 nM) and a detection limit of 0.985 fM. Several signal amplification strategies adopted in this study have effectively improved the performance of the sensor, providing a new avenue for the development of ultrasensitive sensors in the food analysis field.
研究领域:
分析化学
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食品科学
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生物传感技术
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2022
·
IF 9.221
Hui Zhou,Hui Shen,Baoting Dou,Qiumei Feng,Xiguang Han,Po Wang
Methylated DNA is a promising epigenetic biomarker for diseases diagnosis, therapy monitoring, and prognosis. Here, we developed a sensitive ratiometric electrochemical biosensor for the detection of methylated DNA based on the design of multistep DNA amplification circuits. The methylated target DNA firstly initiates the DNAzyme-assisted amplification circuit and produces a number of single-stranded triggers that catalyze the subsequent double-step catalytic hairpin assembly circuits to release numerous four-way DNA junctions. The four-way DNA junctions displace the methylene blue (MB)-modified signal probes for intercalating doxorubicin (DOX) molecules on the sensing interface, resulting in a significantly increased value of i DOX / i MB to achieve sensitive monitoring of methylated DNA. Taking advantage of the significant signal amplification of multistep DNA amplification circuits, the proposed method exhibited high sensitivity for target analysis with a detection limit of 4 aM and a dynamic linear range of 10 aM to 20 pM. Moreover, the proof-of-concept application of the sensing platform was investigated, suggesting that the system is promising for potential applications in early cancer diagnosis and the fundamental research of epigenetics.
研究领域:
分子生物学
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医学检测
·
纳米生物技术
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2023
·
IF 6.1
Chengjie Duan,Yanheng Yao,Wenting Cheng,Yan Chen,Jin Jiao,Yang Xiang
Aptazyme is a chimera of functional nucleic acids , which integrates recognition and amplification elements to simplify the assay process and improve sensing efficiency. However, its application may be limited by signal leakage. In this work, we innovatively integrate the AβO aptamer and an MNAzyme (multicomponent nucleic acid enzyme) for highly efficient detection of AβO. The aptamer and half of the MNAzyme are positioned at one strand, and the other half of the MNAzyme is integrated with a toehold sequence. These two sequences cannot hybridize to activate the MNAzyme until the target is added. The background signal is significantly reduced by the split format and the secondary structure of DNA probes formed in the absence of the target. The proposed aptazyme can not only achieve amplification through enzymatic catalysis but also greatly improve the efficiency of signal transduction and output. We systematically investigated the influence of different DNA probes on the detection performance, and the optimized aptazyme can detect as low as 26.5 pM targets in 1h. The stability of this method was also investigated by detection targets in real biological samples.
研究领域:
生物化学
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分子生物学
·
神经科学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）