SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Yuqing Gan,Xi Long,Zan Gong,Panpan Yuan,Yalan Tang,Shian Zhong,Yanjing Yang
Given the significant threat to human health posed by Staphylococcus aureus , rapid and accurate quantification of S. aureus is essential to prevent and diagnose foodborne diseases and to ensure the safety of food consumption. Here, we developed a novel, simple，and label-free biosensor for quantitatively detecting S . aureus by integrating an allosteric aptamer probe and a rolling circle amplification technique. Pathogen S . aureus-specific binding could induce the structural change of the allosteric aptamer beacon to expose the primer for the padlock probe. Then, the primer could trigger the RCA reaction to produce a long-chain singles-tranded DNA harboring repeating Thioflavin T (ThT) aptamer, leading to powerful fluorescence signals. The assay demonstrates sensitivity, specificity, and accuracy, showing its potential as an alternative tool for pathogen detection in different fields, such as food safety monitoring.
研究领域:
微生物学
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生物技术
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食品科学
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分析化学
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2018
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IF 8.173
Kangyao Zhang,Shuzhen Lv,Minghua Lu,Dianping Tang
In this work, a new “signal-on” split-type photoelectrochemical (PEC) sensing platform for prostate-specific antigen (PSA) detection was successfully constructed using p-type Cu-doped Zn 0.3 Cd 0.7 S as the photosensitive semiconductor material and target-triggered rolling circle amplification (RCA) for signal amplification . The signal derived from Cu-doped Zn 0.3 Cd 0.7 S was amplified by hemin/G-quadruplex. Upon target PSA introduction, the aptamer-primer probe (apt-pri) was captured by capture antibody-conjugated magnetic bead (MB-mAb) to form the sandwiched MB-mAb/PSA/apt-pri. The complex could initiate the RCA reaction to produce a long single-stranded DNA that provided binding sites for G-rich DNA and to form long single-stranded DNA/G-quadruplex/hemin. Upon the addition of exonuclease III (Exo III), the hemin/G-quadruplex immobilized on the RCA long product could be released by the digestion of Exo III. The hemin/G-quadruplex complexes in this study were used as efficient electron acceptors to neutralize the photoelectrons generated from the semiconductor and hindered the recombination of charges, thus enhancing the photocurrent . Under the optimum conditions, the developed sensing system displayed a good analytical performance with a linear range of 0.05–40 ng mL −1 PSA and a detection limit of 16.3 pg mL −1 . Furthermore, good selectivity , high anti-interference ability, satisfactory reproducibility, and good accuracy were also achieved. These prominent analytical properties revealed that our strategy might be a potential and reliable tool for the detection of PSA.
研究领域:
生物化学
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分析化学
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生物传感器
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癌症诊断
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Lulu Dong,Jianhua Wang,Haiqin Li,Yinrong Yang,Xiaochun Li
A low-background paper platform enables cascaded RCA-based ratiometric sensing. • Blue CQDs provide self-calibrated and reliable miRNA-21 quantification. • Smartphone-assisted portable POCT enables noninvasive urinary miRNA analysis. The non-invasive detection of microRNA-21 (miRNA-21), a promising biomarker for bladder cancer, is challenged by its low abundance in urine and the lack of portable, sensitive analytical methods. Herein, we report an integrated smartphone-based paper microfluidic platform that combines rolling circle amplification (RCA), DMSN-based fluorescence enhancement, carbon quantum dots (CQD) -assisted ratiometric fluorescence detection, and smartphone imaging into a portable analytical system for quantitative miRNA-21 detection. A ratiometric fluorescence strategy is employed using blue-emitting CQDs covalently immobilized on glass fiber filter paper as an internal reference, while the RCA product is detected by a DMSN-coated QD probe. Under optimal conditions, the method achieves a limit of detection (LOD) of 1.06 nM for miRNA-21 with a linear range of 0.25–200 nM (R 2 = 0.9891). The assay exhibits excellent specificity against mismatched sequences and other miRNAs, and spike-recovery tests in artificial urine yield recoveries of 88.5–120.3% with relative standard deviations below 15%. The platform shows good batch-to-batch reproducibility (RSD < 7%). Spike recovery tests using clinical urine samples from healthy individuals yielded recoveries of 86.97%–108.5%, confirming the method's high accuracy and reliability in complex urine matrices. Integrated with a 3D-printed portable device and a custom smartphone application, this paper-based sensing platform provides a practical, low-cost, and portable analytical platform for point-of-care testing (POCT) of miRNA-21. The graphical abstract was prepared using SolidWorks, Cinema 4D (C4D), and Microsoft PowerPoint. No AI-assisted tools were used in the generation, preparation, or editing of the figures or graphical abstract. Download: Download high-res image (158KB) Download: Download full-size image
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DOI
高影响因子
ACS Applied Materials & Interfaces
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2026
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IF 7.8
Yang Hongqun,Li Longhao,Niu Mu,Wang Hong,Zhang Yingwei
The development of highly sensitive and reliable biosensors is essential for the early diagnosis and therapeutic monitoring of cancer. Herein, we report a dual-mode fluorescence–colorimetric biosensing platform for the ultrasensitive detection of miRNA-21, integrating AuNP-supported dual-stage rolling circle amplification (RCA) with an RCA-generated multipedal binding-and-cleavage mechanism. The sensing hierarchy is initiated by target-dependent activation of the primary RCA on the gold nanoparticle (AuNP) surface, producing long DNA strands with multiple repeated recognition domains that function as numerous walker “legs” and form a DNA-rich nanogel matrix. Through these repeated domains, the RCA-generated multipedal walker repeatedly and multivalently binds fluorophore-labeled hairpin substrates on the AuNP surface, thereby facilitating APE1-mediated substrate cleavage and fluorescence recovery. The APE1-generated cleavage fragments subsequently initiate the secondary RCA following the addition of the T2 template. The two successive RCA reactions ultimately produce an Au@RCA DNA nanogel containing abundant G-quadruplex/hemin DNAzymes that catalyze a visually detectable color change. By integrating the multipedal walker-mediated multivalent binding-and-cleavage process with dual-stage RCA, the platform enables dual-mode fluorescence–colorimetric detection with efficient signal amplification and low background, achieving limits of detection of 9.854 and 790 fM for the fluorescence and colorimetric modes, respectively. Furthermore, the two complementary outputs provide independent signal readouts for cross-validation. The biosensor demonstrated excellent specificity and distinguished the endogenous responses of MCF-7, HeLa, and HEK293T cell lysates. These results support the potential of the platform for ultrasensitive tumor-associated biomarker detection, while the cell-lysate experiments represent a preliminary proof-of-concept evaluation in complex biological matrices.
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DOI
PubMed
高影响因子
International Journal of Nanomedicine
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2026
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IF 8.7
Jian Tao,Binbin Zeng,Caili Bi,Minyi Xu,Xiaxian Gu,Dingding Jing,Jun Sun
Purpose Ultra-sensitive detection of exosomal miR-21 is crucial for the early diagnosis of colorectal cancer. This study aims to develop a cascade amplification platform based on surface-enhanced Raman scattering (SERS), combining rolling circle amplification (RCA) and CRISPR/Cas12a, to enable ultra-sensitive quantitative detection of exosomal miR-21.Methods A SERS substrate was constructed using streptavidin-modified concave gold nanocubes (CGNs), and gold double-stranded probes loaded with Raman reporter molecules were used as SERS probes. The two components are controllably linked via a single-stranded DNA molecule with a thiol group at one end and biotin at the other, forming a stable detection system. Target-triggered RCA generates a repeat sequence, which, under the mediation of crRNA, activates Cas12a cleavage, causing the probe to be released from the substrate surface and resulting in a significant attenuation of the SERS signal.Results This platform achieves three-stage cascading signal amplification via RCA, CRISPR/Cas12a, and SERS, with a detection limit at the femto-mole (fM) level and a total detection time of approximately 80 minutes. The results were consistent with those of qRT-PCR, demonstrating high sensitivity, high specificity, and reliability.Conclusion This study proposes an RCA-CRISPR-SERS cascade amplification platform that provides a highly sensitive and specific method for detecting miR-21 in exosomes, offering potential clinical value for the early diagnosis and dynamic monitoring of colorectal cancer.
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高影响因子
RSC Advances
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2019
·Vol. 9
·期 66
·38298-38308
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IF 3.049
Xiaojun Xu,Shu Xing,Mengjia Xu,Pan Fu,Tingting Gao,Xiaokang Zhang,Yang Zhao,Chao Zhao
Screening epidermal growth factor receptor (EGFR) mutations, especially deletions, is essential for diagnosis of non-small cell lung cancer (NSCLC) and also critical to inform treatment decisions for NSCLC patients. Here, we demonstrated a facile peptide nucleic acid (PNA) microarray-based fluorometric method for sensitive and specific detection of EGFR mutation, using rolling circle amplification (RCA), graphene oxide (GO), and a fluorescently-labeled detection probe (F-DP). First, the EGFR gene sequence was efficiently captured by the label-free PNA probe which was attached on the surface of a 96-well plate. And then, the EGFR mutation sequence was specifically amplified by RCA using the circular DNA, which was formed by the ligation of the padlock probe when hybridizing with the EGFR mutation, as a template. The single-stranded RCA product (RCAP) was then sensitively detected with the F-DP and GO system. This method has a detection limit of 0.3 pM for EGFR mutation and a high discrimination capability to target EGFR mutation against EGFR wildtype. The use of a PNA microarray and a fluorescence quenching platform make this system quite suitable for high-throughput analysis of EGFR mutations in resource-limited settings without the need of costly and cumbersome equipment. Furthermore, this detection system provides a novel way for the diagnosis of other diseases that are caused by gene deletion mutations.
研究领域:
分子生物学
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癌症诊断
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
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IF 12.545
Jialin Zhang,Yaqi Fan,Jinhui Li,Bin Huang,Herui Wen,Jiali Ren
The key factor to control the incidence rate of diseases caused by bacteria is rapid detection and early diagnosis. Herein, we proposed a new electrochemical bacterial sensor by coupling DNA walking and rolling circle amplification (RCA) reaction-assisted “silver-link” crossing electrode. Staphylococcus aureus ( S. aureus ) was detected using this proof-of concept strategy. Aptamer/DNA walker and auxiliary sequence (AS)/RCA reaction probe (RP) duplexes were modified on the electrode surface . The binding of S. aureus with its aptamer caused the disintegration of aptamer/DNA walker and released DNA walker. With the help of Exo III, DNA walker moved along the electrode surface and AS in AS/RP duplex was continuously digested to release RP. By introducing phi29 DNA polymerase, RCA reaction was performed using RP as the reaction primer to form long single-strand RCA extension products between the electrodes. The “silver-link” crossing electrode was formed by metallization of “gene-link”, significant conductivity was thus acquired for bacteria detection . The limit of detection (LOD) was 10 CFU/mL and detection time was 2 h. The proposed sensor has high efficiency, good stability and low background signal, human serum and milk samples were successfully detected, which emerged a promising potential in the food monitoring and clinical diagnosis.
研究领域:
生物传感器
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分子诊断
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食品监测
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临床诊断
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2019
·
IF 5.256
Guangyang Xiao,Beibei Chen,Man He,Bin Hu
Avian influenza virus (AIVs), hosted in poultry, are the pathogens of many poultry diseases and human infections, which bring huge losses to the poultry breeding industry and huge panic to society. Therefore, it is of great significance to establish accurate and sensitive detection methods for AIVs. In this work, a dual-mode detection method based on immuno-rolling circle amplification (immuno-RCA) and quantum dots (QDs) labeling for inductively coupled plasma mass spectrometry (ICP-MS) and fluorescence detection of H9N2 AIV was developed. The dual-mode detection of the QDs by ICP-MS and fluorescence is used to achieve mutual verification within the analysis results, thus improving the accuracy of the method. With the immuno-RCA, the sensitivity of the method was increased by two orders of magnitude. The limit of detection of the proposed method is 17 ng L −1 and 61 ng L −1 , and the linear range of the proposed method is 0.05–5 ng mL −1 and 0.1–5 ng mL −1 with ICP-MS and fluorescence detection, respectively. The relative standard deviation (n = 7) is 4.9% with ICP-MS detection and 3.1% with fluorescence detection. Furthermore, the proposed method was applied to the analysis of chicken serum samples, no significant different was found for two modes detection and the recoveries of the spiking experiments are acceptable, indicating that the method has good practical potential for real sample analysis.
研究领域:
分析化学
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病毒学
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生物技术
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DOI
PubMed
高影响因子
Scientific Reports
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2018
·Vol. 8
·期 1
·1-9
·
IF 4.122
Yuan Dandan,Ding Lairong,Sun Zhaomei,Li Xuemei
Here, we report a novel magnetic resonance imaging (MRI)/fluorescence bimodal amplification platform for the detection of glutathione (GSH) on the basis of redoxable manganese dioxide (MnO 2 ) nanosheets, which can be readily applied as a DNA nanocarrier, fluorescence quencher, and intracellular GSH-activated MRI contrast agent. The binding of aptamers that absorbed on the MnO 2 nanosheets to their target can facilitating the endocytosis of target-nanoprobes. Once endocytosed, the MnO 2 nanosheets can react with cellular GSH, resulting in the disintegration of nanosheets to generate plenty of Mn 2+ ions for MRI and releases the primers which were adsorbed on the MnO 2 nanosheets. Then the rolling circle amplification (RCA) reaction was initiated to amplify the fluorescence signal. In addition, after treatment with GSH, the MnO 2 nanosheets were reduced and then most of the fluorescence was recovered. Therefore, this MnO 2 nanoprobe exhibits excellent selectivity, suggesting a potential detection platform for analyzing the glutathione level in cells.
研究领域:
生物医学工程
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纳米技术
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癌症研究
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分子成像
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
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2022
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IF 3.191
Ping Wang,Yaoyao Xie,Huikai Ma,Jiancheng Liu,Chenxiang Liu,Wei Feng,Shoumin Xi
A novel ratiometric electrochemical aptasensor was proposed for carcinoembryonic antigen (CEA) detection based on exonuclease III (Exo III)-assisted recycling and rolling circle amplification (RCA) strategies. A thiolated ferrocene-labeled hairpin probe 2 (Fc-HP2) was fixed on the gold nanoparticles (AuNPs)-modified gold electrode (AuE) surface through Au–S bonds. The presence of CEA led to the release of trigger, which hybridized with the 3′-protruding of hairpin probe 1 (HP1) and triggered the Exo III cleavage reaction, accompanied by the releasing of trigger and generation of new DNA fragment which was used for the successive hybridization with Fc-HP2. After the Exo III cleavage process, the remaining Fc-HP2 fragments hybridized as primers with the RCA template to initiate the RCA process, and long single-stranded polynucleotides were produced for methylene blue (MB) binding. Such changes resulted in the signal of Fc (I Fc ) decreased and that of MB (I MB ) increased, achieving a linear relationship between I MB /I Fc and logarithm of CEA concentrations ranging from 1.0 pg mL −1 to 100.0 ng mL −1 with a detection limit of 0.59 pg mL −1 . Additionally, the developed aptasensor had been successfully applied to detect CEA in human serum samples. Therefore, the proposed strategy might provide a new platform for clinical detections of CEA.
研究领域:
生物化学
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分析化学
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生物医学工程
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2022
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IF 6.911
Mengyang Zhou,Haoyu Wang,Chao Li,Chao Yan,Panzhu Qin,Lin Huang
Single base mutations detection is crucial for the diagnosis and treatment of cancer. However, the current methods with poor selectivity and sensitivity required large instruments, which are difficult to meet clinical demands. Herein, we develop a CRISPR/Cas9 based visual colorimetric platform to specifically detect all single base mutations. In this strategy, the Recombinase Polymerase Amplification (RPA) was firstly used to amplify the target, and introduced the PAM site in the target DNA sequence by designing the point mutation primer, thus achieving detection for all single base mutations by the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated specific recognition. With the help of CRISPR/Cas9 system, those RPA products can release single strand DNA to hybridize with the padlock probe and trigger rolling circle amplification (RCA). Based on the magnetic separation, HRP-gold nanoparticles complex (hGNPs) and biotin modified probe (Bio-probe) were further used to achieve enhanced visual variations assay by hybridizing with RCA products. Benefiting from the RPA assisted triple signal amplification, this method not only showed enhanced sensitivity with a limit of detection (LOD) as low as 0.2 fM and 0.01% of KRAS-G12D mutation percentage, but the specificity against KRAS-G12D mutation also be synergistically enhanced by combining the CRISPR/Cas9-mediated specific recognition with the specific T4 ligation reaction of RCA system. Furthermore, this system has been successfully used to visually detect genome in serum, suggesting its great potential for point-of-care diagnosis in clinical.
研究领域:
分子生物学
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生物技术
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医学诊断
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癌症研究
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 144
·期 20
·5996-6003
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IF 4.019
Mei-Ling Gao,Bin-Cheng Yin,Bang-Ce Ye
As exosomes have been playing an increasingly important role in the diagnosis, treatment and prognosis of diseases, the analysis of exosome contents becomes more crucial. Therefore, the development of a cost-effective and simple exosome separation method that achieves high purity is urgently needed, and it is vital for further research in cancer. In this work, we constructed a DNA-AuNP-based satellite network which integrates low-speed centrifugal exosome isolation, detection and protein analysis. The rolling circle amplification (RCA) reaction is used to produce a long-chain DNA hairpin structure comprising a plurality of functional domains, such as CD63 aptamer sequences, linker sequences, and spacer sequences with complementary base pairs to form a hairpin structure. When the CD63 aptamers bind to exosomes, the hairpin structure changes its conformation, exposing the linker sequences (AuNP binding sequence). Then the probe on the surface of AuNPs combines with the long-chain DNA by the toehold-mediated strand displacement reaction, releasing the fluorescent labeled complementary probe as the detection signal and simultaneously forming the DNA-AuNP-based satellite network. Thus, exosomes can be isolated by low-speed centrifugation. The formation of the DNA-AuNP-based satellite network was confirmed by transmission electron microscopy and confocal fluorescence microscopy. In addition, we established a standard curve for exosome detection which showed good linearity of the fluorescence ratio vs. log(exosome concentration). LC/MS for protein profiling of the captured exosomes demonstrated that our method has potential application in the field of exosome research.
研究领域:
生物技术
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纳米技术
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分子生物学
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癌症研究
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）