Biosensors-Basel
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2022
·Vol. 12
·期 11
·989
·
IF 5.743
Aiting Cai,Luxia Yang,Xiaoxia Kang,Jinxia Liu,Feng Wang,Haiwei Ji,Qi Wang,Mingmin Wu,Guo Li,Xiaobo Zhou,Yuling Qin,Li Wu
An electrochemical-DNA (E-DNA) sensor was constructed by using DNA metallization to produce an electrochemical signal reporter in situ and hybridization chain reaction (HCR) as signal amplification strategy. The cyclic voltammetry (CV) technique was used to characterize the electrochemical solid-state Ag/AgCl process. Moreover, the enzyme cleavage technique was introduced to reduce background signals and further improve recognition accuracy. On the basis of these techniques, the as-prepared E-DNA sensor exhibited superior sensing performance for trace ctDNA analysis with a detection range of 0.5 fM to 10 pM and a detection limit of 7 aM. The proposed E-DNA sensor also displayed excellent selectivity, satisfied repeatability and stability, and had good recovery, all of which supports its potential applications for future clinical sample analysis.Keywords:DNA metallization;ctDNA;HCR;Exo Ι;E-DNA sensor
研究领域:
生物化学
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分析化学
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纳米技术
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分子生物学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2025
·
IF 6
Mimi Li,Lina Wang,Yue Hu,Yi Liu,Shuang Xu,Kexing Peng,Chenghong Li,Hui Huang,Lichao Fang,Lulu Li,Huamin Liu,Xiaolong Wang,Junsong Zheng
Sensitive and efficient detection of RNA methylation sites is considered an integral part of epigenetic assessment. Contrary to previous studies on the detection of RNA m5C or m6A methylation alone, this study innovatively designed a biosensor capable of detecting RNA m5C or m6A methylation at the same time. Substances with specific electrochemical reactions such as ferrocene (FC) or methylene blue (MB) were modified onto the hairpin probe. When the hairpin probe was activated under specific conditions, it triggered a dendritic nonlinear hybridization chain reaction (HCR), which resulted in signal amplification. Gold (Au) and iron tetraoxide (Fe 3 O 4 ) composite nanomaterials were employed as the linking materials: the carboxylated ends of Fe 3 O 4 were connected to an antibody that specifically recognizes the m5C and m6A methylation sites, while Au nanoparticle ends adhering to the carboxylated Fe 3 O 4 surface are connected to the HCR enhanced signal amplifier. The m5C antibody was linked to Fc-containing HCR amplification products by this material, and similarly, the m6A antibody was linked to MB-containing amplifiers. Thus, the dandelion complex, a multifunctional body with methyl recognition and signal amplification, was formed. The capture probe immobilized on the surface of the gold electrode by Au–S recognized the target RNA sequence by base complementary pairing. Upon addition of the dandelion complex, due to its multi-functionality, the amplified signals were carried to the electrode by the antigen-antibody recognition mechanism, generating a current signal. The positions and heights of the current signal peaks enabled rapid determination of target RNA methyl modification sites and their abundance. The detection limits of this biosensor for RNA m5C and m6A were 4.68 × 10 −16 M and 1.10 × 10 −15 M, respectively, and the linear range was from 10 −15 mol/L to 10 −8 mol/L. The sensor developed in this study has the advantages of cost-effectiveness, ease of fabrication, and fast response time, and has the potential to be promoted for RNA methylation detection.
研究领域:
分子生物学
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生物传感技术
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表观遗传学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Xiaokun Wang,Hao Ding,Yinghui Sun,Yanling Ma,Guoqing Wang,Jiadong Chen,Jaebum Choo,Lingxin Chen
To address the need for highly sensitive and reliable detection of trace ochratoxin A (OTA) in food matrices, we developed a ratiometric fluorescent aptasensor by integrating CRISPR/Cas12a, hybridization chain reaction (HCR), and horseradish peroxidase (HRP)-induced inner filter effect (IFE). The mechanism involves OTA releasing an activator that initiates CRISPR/Cas12a trans-cleavage, blocking HCR assembly. This reduces HRP levels, limiting the conversion of o -phenylenediamine (OPD) to fluorescent 2,3-diaminophenazine (DAP) (emitting at 562 nm) while maintaining strong emission from 2-amino terephthalic acid (BDC-NH 2 ) at 426 nm. The F 426 /F 562 ratio serves as a “signal-on” indicator, enabling sensitive OTA detection over 0.1 pM to 10 nM, with a detection limit of 0.0417 pM. The method exhibits excellent reproducibility, with intra-day and inter-day relative standard deviations (RSDs) of 1.91 %–3.87 % and 1.79 %, respectively, along with recovery rates of 90.1 %–110.6 % in real samples. These advantages highlight its significant potential for CRISPR/Cas-based OTA detection.
研究领域:
食品安全
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Hua-Jie Chen,Yusi Hu,Peiyu Yao,Di Ning,Yu-Peng Zhang,Zhi-Gang Wang,Shu-Lin Liu,Dai-Wen Pang
Cardiovascular disease is one of the main causes of death in the world, which is closely associated with dyslipidemia. Dyslipidaemia is usually manifested as a relatively higher level of low-density lipoprotein (LDL) and lower level of high-density lipoprotein (HDL). Thus, the quantitative detection of the LDL and HDL particles is of great importance to predict the risk of cardiovascular diseases. However, the traditional methods can only indirectly reflect the HDL/LDL particle concentrations by detecting the cholesterol or proteins in HDL/LDL particles and are always laborious and time-consuming. Thus, the accurate and efficient approach for the detection of intact HDL and LDL particles is still lacking so far. We developed an enzyme- and isolation-free method to measure the concentration of HDL and LDL based on DNAzyme and hybridization chain reaction (HCR)-based signal amplification. This method can be used to directly and accurately detect the concentration of “actual” HDL and LDL particles instead of the cholesterol in HDL and LDL, with limits of detection of 10 and 30 mg/dL, respectively, which also satisfied the lipoprotein analysis in clinical samples. Therefore, this HCR–DNAzyme platform has great potential in clinical applications and health management.
研究领域:
医学诊断
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分子生物学
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生物技术
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DOI
PubMed
高影响因子
Analytical Methods
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2022
·Vol. 15
·期 1
·99-108
·
IF 3.532
Hongyan Zhang,Siying Ye,Lishan Huang,Shen Fan,Weiwei Mao,Yijin Hu,Yuyan Yu,Fengfu Fu
Aflatoxin B1 (AFB1) is a highly toxic mycotoxin, which causes severe acute or cumulative poisoning. Therefore, it is important to develop sensitive and selective detection methods for AFB1 for the safety of food and medicinal herbs. Herein, we have developed a “signal-on” electrochemical aptasensor based on the high specificity of the aptamer and hybridization chain reaction (HCR) biological amplification for AFB1 detection. In this work, thiol-modified complementary DNA (cDNA) immobilized on the surface of a gold electrode (GE) served as an initiator DNA. When AFB1 was present, it competed with the cDNA for binding to the aptamers, which resulted in the detaching of aptamers from the cDNA–aptamer duplexes. Then, the single-stranded cDNA acted as an initiator to trigger the HCR signal amplification. Therefore, long double-stranded DNA (dsDNA) products were produced, which could load large amounts of methylene blue (MB) molecules to generate a distinct electrochemical signal. Under the optimized conditions, the proposed electrochemical aptasensor achieved the ultrasensitive detection of AFB1 with a linear detection range of 0.01–100 pg mL−1, and a limit of detection (LOD) down to 2.84 fg mL−1. Furthermore, the electrochemical aptasensor was successfully applied for detecting AFB1 in corn and two kinds of traditional Chinese medicine samples, indicating the potential value for AFB1 detection in practical samples.
研究领域:
分析化学
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毒理学
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食品安全
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生物技术
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2025
·
IF 5.1
Guobin Huang,Weiting Xu,Jinxin Chi,Chenchen Lin,Jiabin Wang,Xucong Lin
Environmental smog pollution induces chronic obstructive pulmonary disease (COPD), and causes the urgent need of timely, safe and quantitative testing for risk assessment. The miRNA-based bioanalysis offers a non-invasive access to rapid COPD screening, while challenges remain in accurately identifying miRNAs from complex matrices. Herein, a novel bionic aptasensor has been pioneered for meeting the specific and sensitive detection of the miR-155 in serum for COPD diagnosis, integrating tetrahedral framework nucleic acid (tFNA) confined affinity recognition and enzyme-free hybridization chain reaction (HCR) amplification strategy. Precision-designed tFNAs were fabricated and hybridized with FAM-labeled cDNAs, enabling dual-mode recognition mechanism including spatially confined aptamer recognition and bionic molecular sieve size-selective action. Proteins and pre-miRNAs were excluded, and the target microRNA-155 was specifically captured. This triggered a structural switch of tFNAs and activated cDNA-mediated HCR, assembling double-stranded DNA and achieving exponential fluorescence enhancement synergistically produced by SYBR Green I (SGI) and FAM. The optimal limit of detection as 53 fM for miRNA-155 was achieved with laser-induced fluorescence (LIF). Applied to COPD diagnosis, the detections of trace miRNA-155 were satisfactory and the recovery yields were ranged in 96.4 ± 5.2 % ∼106.2 ± 2.1 % ( n = 3) in the serum of COPD patients. Smoke-exposed model rats were also analyzed, and the miRNA-155 was achieved with a significant concentration up-regulation to light the COPD. It offers a high-performance protocol for meeting the non-invasive rapid screening of COPD, further providing promising technologies for evaluating safety issues of environmentally induced diseases.
研究领域:
生物传感器
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分子诊断学
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环境健康
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RNA生物学
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肺部疾病研究
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DOI
高影响因子
Analytical Methods
·
2023
·Vol. 16
·期 4
·496-502
·
IF 3.1
Hangli Yan,Linghao Wu,Jingyu Wang,Yi Zheng,Fengxia Zhao,Qinqin Bai,Hongmei Hu,Hao Liang,Xiangheng Niu
The detection of foodborne pathogens is crucial for food hygiene regulation and disease diagnosis. Colorimetry has become one of the main analytical methods in studying foodborne pathogens due to its advantages of visualization, low cost, simple operation, and no complex instrument. However, the low sensitivity limits its applications in early identification and on-site detection for trace analytes. In order to overcome such a limitation, herein we propose a joint strategy featuring dual signal amplification based on the hybridization chain reaction (HCR) and DNA-enhanced peroxidase-like activity of gold nanoparticles (AuNPs) for the sensitive visual detection of Escherichia coli. Target bacteria bound specifically to the aptamer domain in the capture hairpin probe, exposing the trigger domain for HCR and forming the extended double-stranded DNA (dsDNA) structures. The peroxidase-like catalytic capacity of AuNPs can be enhanced significantly by dsDNAs with the sticky ends of dsDNAs being adsorbed on AuNPs and the rigidity of dsDNAs causing the spatial regulation of AuNP concentration. The intensity of the enhancement was linearly related to the number of target bacteria. With the above strategy, the detection limit of our colorimetric method for Escherichia coli was down to 28 CFU mL−1 within a short analytical time (50 min). This study provides a new perspective for the sensitive and visual detection of early bacterial contamination in foods.
研究领域:
食品科学
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生物技术
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分析化学
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微生物学
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DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
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2025
·
IF 4.5
Ai Ting Cai,Qing Ping Fu,Ming Zhong Sun,Hong Mei Chen,Yun Gang Wang,Hui Xiang Jv
The detection of BRAF mutations in circulating tumor DNA is beneficial for early diagnosis and personalized therapeutic approaches of melanoma. However, the sensitive and reliable trace detection of BRAF mutations remains a major challenge. Therefore, in this study, the combination of two isothermal signal amplification techniques (primer exchange reaction (PER) and hybridization chain reaction (HCR)) was proposed to construct an electrochemical biosensor for the efficient and sensitive detection of BRAF mutations. The presence of target triggered PER and HCR sequentially to achieve dual-cascade amplification of positive electrochemical signal feedback. Compared to the detection limit (1.27 pM or 0.23 pM) of single-signal amplification technology (PER or HCR), the detection limit of dual-signal amplification techniques was significantly lower (0.34 fM). In addition, the dual-signal amplification strategy exhibited other excellent sensing properties, such as a wide linear detection range (0.01 pM to 1 nM), excellent specificity, satisfactory reproducibility and stability, and the ability to achieve low-level target detection in diluted serum samples. Based on all of these features, the proposed strategy indicate broad application prospects for detecting low-abundance nucleic acid targets in clinical samples.
研究领域:
生物传感器
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分子诊断
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生物化学
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癌症研究
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分析化学
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遗传学
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电化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·
IF 4.2
Jia Chen,Shengrong Yu,Zhiling Qian,Kangdi He,Bingqian Li,Yuting Cao,Keqi Tang,Shengjia Yu,Yong-Xiang Wu
Rapid, simultaneous, and sensitive detection of biomolecules has important application prospects in disease diagnosis and biomedical research. However, because the content of intracellular endogenous target biomolecules is usually very low, traditional detection methods can't be used for effective detection and imaging, and to enhance the detection sensitivity, signal amplification strategies are frequently required. The hybridization chain reaction (HCR) has been used to detect many disease biomarkers because of its simple operation, good reproducibility, and no enzyme involvement. Although HCR signal amplification methods have been employed to detect and image intracellular biomolecules, there are still false positive signals. Therefore, a target-triggered enzyme-free amplification system (GHCR system) was developed, as a fluorescent AND-gated sensing platform for intracellular target probing. The false positive signals can be well avoided and the accuracy of detection and imaging can be improved by using the design of the AND gate. Two cancer markers, GSH and miR-1246, were used as two orthogonal inputs for the AND gated probe. The AND-gated probe only works when GSH and miR-1246 are the inputs at the same time, and FRET signals can be the output. In addition to the use of AND-gated imaging, FRET-based high-precision ratiometric fluorescence imaging was employed. FRET-based ratiometric fluorescent probes have a higher ability to resist interference from the intracellular environment, they can avoid false positive signals well, and they are expected to have good specificity. Due to the advantages of HCR, AND-gated, and FRET fluorescent probes, the GHCR system exhibited highly efficient AND-gated FRET bioimaging for intracellular endogenous miRNAs with a lower detection limit of 18 pM, which benefits the applications of ratiometric intracellular biosensing and bioimaging and offers a novel concept for advancing the diagnosis and therapeutic strategies in the field of cancer.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
Analytical Methods
·
2026
·
IF 3
Huiling Xue,Bing Zhang,Ying Zhan,Zhihuan Zhao,Jianying Lin,Yan Cheng
In this study, a novel photothermal sensing platform was established for the visual, portable, and quantitative detection of histamine. This approach is built based on a hybridization chain reaction (HCR) as nucleic acid amplification strategy, which is triggered by histamine.CuS@Au nanocomposites were synthesized through a hydrothermal method and utilized as the photothermal transduction material, which displayed excellent photothermal conversion performance, effectively converting histamine concentration into a measurable temperature variation. Under optimized conditions, the temperature difference (ΔT) presented a favorable linear relationship with histamine concentration in a range of 10 pM -10 μM, with a detection limit as low as 0.82 pM. Moreover, the practicality of this method was successfully verified by detecting histamine spiked in fish and shrimp meat samples. The developed sensing platform shows considerable potential for applications in food safety monitoring and public health protection.
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 7
·1-11
·
IF 5.3
Liang Pengda,Lv Bei,Chen Ke,Li Dawei
The combination of CRISPR/Cas12a and functional DNA provides the possibility of constructing biosensors for detecting non-nucleic-acid targets. In the current study, the duplex protospacer adjacent motif (PAM) in the activator of CRISPR/Cas12a was used as a molecular switch, and a sensitive adenosine triphosphate (ATP) detection biosensor was constructed using an allosteric probe-conjugated PAM site formation in hybridization chain reaction (HCR) integrated with the CRISPR/Cas12a system (APF-CRISPR). In the absence of ATP, an aptamer-containing probe (AP) is in a stem-loop structure, which blocks the initiation of HCR. In the presence of ATP, the structure of AP is changed upon ATP binding, resulting in the release of the HCR trigger strand and the production of long duplex DNA with many PAM sites. Since the presence of a duplex PAM site is crucial for triggering the cleavage activity of CRISPR/Cas12a, the ATP-dependent formation of the PAM site in HCR products can initiate the FQ-reporter cleavage, allowing ATP quantification by measuring the fluorescent signals. By optimizing the sequence elements and detection conditions, the aptasensor demonstrated superior detection performance. The limit of detection (LOD) of the assay was estimated to be 1.16 nM, where the standard deviation of the blank was calculated based on six repeated measurements. The dynamic range of the detection was 25–750 nM, and the whole workflow of the assay was approximately 60 min. In addition, the reliability and practicability of the aptasensor were validated by comparing it with a commercially available chemiluminescence kit for ATP detection in serum. Due to its high sensitivity, specificity, and reliable performance, the APF-CRISPR holds great potential in bioanalytical studies for ATP detection. In addition, we have provided a proof-of-principle for constructing a CRISPR/Cas12a-based aptasensor, in which the PAM is utilized to regulate Cas12a cleavage activity. Graphical Abstract
研究领域:
生物化学
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Guizhu Wu,Guangyun Chen,Junjian Li,Zhaoying Liu,Zhanming Li,Lijuan Hou,Zhen Zhang,Ming Li
A tri-mode aptasensor for ultrasensitive detection of ochratoxin A (OTA) was developed, based on two-dimensional hybridization chain reaction (2D-HCR)-guided etching to modulate gold nanobipyramid (AuNB) morphology. The OTA aptamer-functionalized magnetic probe (MB@2D-HCR@HRP) integrated the self-assembly 2D-HCR networks, which had loaded with abundant horseradish peroxidase (HRP) for cascade signal amplification. Following OTA recognition, the released 2D-HCR@HRP could induce the generation of hydroxyl radical (·OH), then perform the AuNB etching. The results of AuNB morphological transformation identified through three readouts: (1) visual multicolor (LOD of 100 pg mL −1 ), (2) surface plasmon resonance peak (LOD of 75.3 pg mL −1 ), (3) photothermal response (LOD of 14.7 pg mL −1 ). Thus, the levels of OTA could directly reflect via triple signals, achieving rapid and ultrasensitive detection within 45 min. The aptasensor exhibited high specificity (cross-reactivity less than 7.22 %) and reliability for spiked and authentic agricultural byproduct samples. Furthermore, the highlights of simplified operation visualization and portability had displayed. This aptasensor had handled advanced strategies to improve the detection performance, which might further promote the practicability and accessibility of biosensing.
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高影响因子
危险品化学品经营许可证（带存储）