TALANTA
·
2024
·
IF 5.6
Lifeng Zhang,Shihua Luo,Rui Fan,Ruixi Li,Wenbin Li,Siting Chen,Fei Lan,Yitong Zhu,Tingting Ji,Ye Zhang,Ling Li
APE1, an essential enzyme for DNA repair, is overexpressed in various cancers and has been identified as a potential biomarker for cancer diagnosis. However, detecting APE1 at low expression levels in the early stage of cancer presents a significant obstacle. Here, we introduced a novel localized Cas12a-based cascade amplification (LCas12a-CA) method. This method confined both the terminal deoxynucleotidyl transferase and the crRNA/Cas12a complex onto the surfaces of gold nanoparticles (AuNPs). This confinement not only boosts the stability of the multiple enzymes but also induces a substrate channeling effect. As a result, it significantly accelerates the reaction rate and enhances the sensitivity of APE1 detection. Upon the addition of APE1, the AP sites within the APE1 primer can be recognized and cleaved by APE1, exposing the 3′-OH ends. In the presence of LCas12a-CA, polyA sequences are generated at 3′-OH ends with the help of TdT and dATP. The sequences directly enter the Cas12a system, activating the trans -cleavage activity of Cas12a, thereby cutting the reporters on the surface of AuNPs and releasing fluorescence. Our platform demonstrates a detection limit (LOD) as low as 2.51 × 10 −6 U/mL, which is more than 60 times lower than that of free Cas12a-CA. Furthermore, the LCas12a-CA exhibits enhanced resistance ability in extreme environments and has been proven effective for the detection of APE1 in clinical samples. Overall, this work offers a promising platform for robust biosensing in cancer diagnosis and prognosis.
研究领域:
分子生物学
·
癌症研究
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Wenting Cheng,Han Pan,Jinhua Chen,Miao He,Zhongyun Wang,Yang Xiang
Exosomal miRNAs, particularly miRNA-21, are promising cancer biomarkers. Current enzyme-dependent detection methods face challenges, such as environmental limitations and high costs. In contrast, enzyme-independent sensors are highly desirable for on-site, miniaturized, and cost-effective miRNA detection. To address these limitations, we developed a nonenzymatic electrochemical sensor featuring a triple-signal amplification system for ultrasensitive detection of miRNA-21. This sensor utilizes cascade toehold-mediated strand displacement reactions to activate molecular machines triggered by target miRNA, generating biotinylated-and-thiol-modified double-stranded DNA for stable immobilization on a gold electrode. Preprepared biotinylated tetrahedron DNA (TDNA)-mediated hybridization chain reaction probes are then linked to the electrode via streptavidin–biotin binding. This amplification process allows for significant DNA duplex immobilization, with electroactive [Ru(NH3)6]3+ (RuHex) adsorbed onto them, producing a robust electrochemical signal. This approach enables accurate detection of miRNA-21 at concentrations as low as 0.43 fM, with a linear range from 1 fM to 1 nM. Clinical testing demonstrates its potential for cancer diagnostics.
研究领域:
生物化学
·
医学诊断
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-19
·
IF 15.7
Ren Tingju,Zhang Yingzhi,Tong Yuxiao,Zhang Qi,Wang Tianhao,Wang Yue,Yang Chunguang,Xu Zhangrun
Glycosylated RNAs (glycoRNAs), a recently discovered class of membrane-associated glyco-molecules, remain poorly understood in function and clinical value due to limited detection methods. Here, we show a dual recognition Förster resonance energy transfer (drFRET) strategy using nucleic acid probes to detect N-acetylneuraminic acid-modified RNAs, enabling sensitive, selective profiling of glycoRNAs on small extracellular vesicles (sEVs) from minimal biofluids (10 μl initial biofluid). Using drFRET, we identify 5 prevalent sEV glycoRNAs derived from 7 cancer cell lines. In a 100-patient cohort (6 cancer types and non-cancer controls), sEV glycoRNA profiles achieve 100% accuracy (95% confidence interval) in distinguishing cancers from non-cancer cases and 89% accuracy in classifying specific cancer types. Furthermore, drFRET reveal that sEV glycoRNAs specifically interact with Siglec proteins and P-selectin, which is critical for sEV cellular internalization. The drFRET strategy provides a versatile and sensitive platform for the imaging and functional analysis of sEV glycoRNAs, with promising implications for clinical applications.
研究领域:
分子生物学
·
肿瘤学
·
临床医学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Interdisciplinary Medicine
·
2025
·
IF 13.6
Qin-Xin Gao,Qian-Qian Wu,Zhi-Ying Dong,Xiao-Yuan Qin,Lin-Yan Zhai,Bao-Ping Xie,Wen-Jun Duan
Increasing studies have demonstrated that PIWI-interacting RNAs (piRNAs) in circulating exosomes can serve as novel molecular biomarkers for tumor liquid biopsy. However, methods for in situ detection of piRNAs encased in exosomes are limited. In this study, we designed a spherical nucleic acid probe named piR-651, which can enter exosomes simply by incubating with them for 2 h and in situ detect piR-651 with a detection limit of 5 × 10 7 particles/μL. Based on this probe, we established a liquid biopsy method for the in situ detection of piR-651 in plasma exosomes. The assay could distinguish the expression levels of piR-651 between 21 breast cancer patients and 22 healthy individuals. The receiver operating characteristic curve shows an area under the curve as 0.9931 and the diagnostic sensitivity and specificity at the best cutoff are 85.7% and 100%, respectively. The probe can also easily perform in situ imaging of piR-651 in living cells. To avoid low sensitivity and kinetics in detecting large-sized PIWI-interacting RNA complexes, we rationally designed the structure and detection scheme of piR-651 probe, which was synthesized by modifying 13-nm gold particles with high-density Anchor-Report DNA duplexes through the butanol dehydration method. The new design of the gold nanoparticle nucleic acid probe can be applied to the fabrication of nucleic acid probes targeting other large-volume nucleic acids for developing more molecular biomarker-based liquid biopsy for cancer diagnosis.
研究领域:
分子生物学
·
肿瘤学
·
生物医学工程
·
遗传学
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2025
·
IF 3.3
Xiaoming Kong,Chao Li,Yang Li,Xueqian Song,Lin Huang
Exosomes, which are recognized as a kind of valuable liquid biopsy biomarker, exhibit significant application potential in cancer diagnosis. Therefore, it is crucial to establish a reliable detection method for their clinical application. In this study, we presented an ultrasensitive aptasensor for the visual detection of exosomes by employing tyramine-assisted dual signal amplification technology. Firstly, we utilized magnetic beads modified with the nucleolin aptamer (MNPs-Aptnucleolin) to capture exosomes. This modification not only enhanced specificity, but also reduced interference of complex sample components. The captured exosomes as a rich source of proteins could bind with the multiple biotinyl-tyramide (Bio-TR) through a catalytic reaction involving horseradish peroxidase (HRP) and H2O2. Secondly, streptavidin-HRP complex-modified gold nanoparticles (GNPs-Str-HRP) as signal amplification probe was introduced to further enhance the detection signal by binding to Bio-TR. Lastly, the addition of 3,3',5,5'-tetramethylbenzidine (TMB) solution induced a visible color change, enabling the quantification of exosome concentration. This dual signal amplification strategy resulted in a low limit of detection (LOD) of 63 particles/μL, and also demonstrated accurate visual diagnosis capabilities for clinical samples. The successful implementation of this approach suggests its potential as a promising tool for point-of-care testing (POCT) in cancer diagnostics.
研究领域:
医学诊断
·
生物化学
·
肿瘤学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JACS Au
·
2025
·
IF 8.7
Weiming Lin,Tao Ding,Die He,Nan Zhang,Haodong Li,Wenjian Luo,Zhongxia Wei,Min Ke,Sisi Jia,Chunhai Fan,Le Liang
Nanodiamonds (NDs) with nitrogen-vacancy (NV) centers are emerging as powerful quantum nanosensors (QNs) in biomedical applications due to their exceptional sensitivity. However, achieving optimal diagnostics performance necessitates both high sensitivity and selectivity; especially in practical biomedical settings, it remains challenging for QNs to provide quantitative analyses when multiple analytes are present. Here, we present a biosensing platform that integrates DNA logic gates (DLGs) with spin-based quantum sensing, termed DLG-QN for ultrasensitive and ultraselective diagnostics. Utilizing an AND DLG, both NDs and magnetic beads (MBs) are functionalized with hairpin DNA strands. In the presence of both miRNA-21 and miRNA-155─key biomarkers overexpressed in cancer─the hairpin DNAs undergo conformational changes that facilitate DNA-guided self-assembly of NDs and MBs, enriching the target signal. Resonant microwave modulation of ND fluorescence emission allows for high signal-to-noise ratio (SNR) detection by separating the signal from background fluorescence via spin-enhanced analysis. This platform demonstrated ultrasensitive and ultraselective detection of miRNA-21 and miRNA-155 with a limit of detection of 19.8 fM, highlighting its potential as a general biosensing strategy for precision diagnostics involving multiple biomarkers.
研究领域:
生物医学工程
·
分子诊断学
·
量子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 8
·1-9
·
IF 5.3
Chen Jinri,Xu Dong,Huang Qiang,Wang Shujun,Li Fuhou,Wu Shaojie,Wang Weixia,Zhou Nandi
Sialyl-Lewis x (SLe x ) is a tetrasugar, which plays an important role in initial inflammation and cancer cell metastasis, and can be used as a marker for cancer diagnosis and prognosis or a therapeutic target. Detecting SLe x from complex biological media remains a significant challenge. Herein, a single-stranded DNA aptamer of SLe x was screened based on the double-stranded DNA library–modified magnetic bead (MB)-SELEX technology. After 14 rounds of screening, 12,639 sequences were obtained and divided into nine families. Three representative sequences were selected based on the number of sequence repeats and Gibbs binding free energy, and the aptamer SLe x -Apt2 with 80 nt length ( K d = 23.01 nM) had the best affinity and relatively high specificity for targeting SLe x . Then, a novel dual-recognition fluorescent biosensor for SLe x -sensitive detection based on aptamer SLe x -Apt2 bio-dots and 3-aminobenzoboric acid–modified MB was developed. This method can detect SLe x as low as 32 μM and has a good linear response in the range 100 μM to 2 mM. It has the advantages of low preparation cost, good targeting, and avoiding the occurrence of false-positive and false-negative detection results, which makes the biosensor more valuable in biological detection and clinical diagnosis. Graphical Abstract
研究领域:
生物化学
·
分子生物学
·
临床医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Mengyi Tian,Youqiang Xing,Wei Wang,Ze Wu,Peng Huang,Bingjue Li,Lei Liu
Accurate detection of microRNA concentrations is essential for early cancer diagnosis and monitoring. Existing methods typically provide a linear response within a limited concentration range, which hinders their precise clinical application. This study presents a novel sensing approach using the fluorescence properties of nitrogen vacancy (NV) centers. Magnetic particles capture and enrich fluorescent nanodiamonds (FNDs), and effectively address signal inconsistencies due to variations in FND particles by applying a voltage that regulates the NV center fluorescence. Our method achieves a linear response to microRNAs across a broad concentration range (10 fM to 1 μM), with the determination coefficient (R²) of the fluorescence signal, enhanced from 0.8711 to 0.9951 through voltage modulation. Additionally, optimizing experimental conditions and enhancing FND fluorescence increased the signal intensity, achieving a minimum detection limit of 7.17 fM. The combination of sensitive detection and an extensive linear range suggests the potential of this strategy as a universal detection method for the early diagnosis of various diseases.
研究领域:
生物医学工程
·
分子生物学
·
量子传感
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Jixuan Han,Xiaoran Liu,Ying Yan,Jingwen Ge,Han Zhao,Chen Wang,Guohong Song,Ling Zhu,Yanlian Yang
Extracellular vesicle miRNAs (EV-miRNAs) are strongly linked to cancer progression, metastasis, and drug resistance, making them promising biomarkers for precision diagnosis. However, the clinical potential of EV-miRNA-based liquid biopsies is hindered by the low abundance of EV-miRNAs and the tedious detection procedure including EV separation, purification and miRNA quantification. Here, we develop a novel one-step catalytic hairpin assembly (CHA)-based fluorescent assay for sensitive detection of EV-miRNAs assisted with DNA-mediated membrane fusion (DMF) and DNA tetrahedron (DT) (DMF-DT-CHA). This DMF-DT-CHA assay facilitates the membrane fusion between liposome and EV through interactions between DNAs for DT-CHA probe delivery into EVs, and followed by DT-CHA, which recognizes target miRNAs to initiate non-enzymatic signal amplification via CHA-based fluorescence emission. We analyzed EVs from three breast cancer cell sources using DMF-DT-CHA and the results were consistent with qPCR and the platform achieved the limit of detection (LoD) of 0.24 fM for EV-miRNAs. We performed a clinical evaluation of the DMF-DT-CHA assay platform. Recipient operating characteristic curves (ROCs) showed that the DMF-DT-CHA platform was an excellent classifier for distinguishing breast cancer (BC) patients from healthy donors and breast cancer patients from benign breast nodule patients, with area under the curve (AUC) values of 0.850 and 0.835, respectively, as well as an accuracy of 81.1% in response to treatment for triple-negative breast cancer. DMF-CT-CHA assay platform has clinical potential for cancer diagnosis and treatment monitoring.
研究领域:
生物化学
·
分子生物学
·
临床医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Shizeng Pei,Haoyang Li,Linfeng Chen,Gang Nie,Huiling Wang,Chunrong Liu,Caihong Zhang
Abnormal lipid droplets (LDs) are known to be intimately bound with the occurrence and development of cancer, allowing LDs to be critical biomarkers for cancers. Aggregation-induced emission luminogens (AIEgens), with efficient reactive oxygen species (ROS) production performance, are prime photosensitizers (PSs) for photodynamic therapy (PDT) with imaging. Therefore, the development of dual-functional fluorescent probes with aggregation-induced emission (AIE) characteristics that enable both simultaneous LD monitoring and imaging-guided PDT is essential for concurrent cancer diagnosis and treatment. Herein, we reported the development of a novel LD-targeting fluorescent probe (TDTI) with AIE performance, which was expected to realize the integration of cancer diagnosis through LD visualization and cancer treatment via PDT. We demonstrated that TDTI, with typical AIE characteristics and excellent photostability, could target LDs with high specificity, which enables the dynamic tracking of LDs in living cells, specific imaging of LDs in zebrafish, and the differentiation of cancer cells from normal cells for cancer diagnosis. Meanwhile, TDTI exhibited fast ROS generation ability (achieving equilibrium within 60 s) under white light irradiation (10 mW/cm2). The cell apoptosis assay revealed that TDTI effectively induced growth inhibition and apoptosis of HeLa cells. Further, the results of PDT in vivo indicated that TDTI had a good antitumor effect on the tumor-bearing mice model. Collectively, these results highlight the potential utility of the dual-functional fluorescent probe TDTI in the integrated diagnosis and treatment of cancer.
研究领域:
生物医学
·
肿瘤学
·
分子影像学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Alexandria Engineering Journal
·
2024
·
IF 6.2
Zhenlu Wang,Yanlong Shi,Zihan Xu,Mengmeng Sun,Xin Shen,Kun Wu,Mingxin Yu,Le Zhang,Guohua Yu
This study presents a cancer diagnostics method through the development of a graphitic carbon nitride (g-C 3 N 4 )-based electrochemical sensor for detecting circulating tumor cells (CTCs) in blood. Utilizing the unique properties of g-C 3 N 4 nanosheets combined with gold nanoparticles functionalized with DNA aptamers, the sensor exhibits remarkable sensitivity and selectivity in CTC detection. The aptamers specifically target cell surface biomarkers, enabling the precise capture and quantification of CTCs. The sensor's detection limit is an impressive 5 cells/mL, with a broad detection range from 10 to 10 5 cells/mL. This high-performance sensing platform not only achieves rapid and accurate quantification of CTCs but also demonstrates robustness against interference from other cell types, making it highly suitable for clinical applications. The optimized conditions for aptamer-functionalized nanoparticle concentration and incubation time ensure the sensor's efficacy, as validated in spiked blood samples. This work provides a practical and efficient approach for non-invasive cancer diagnostics, offering significant improvements over existing methods in terms of simplicity, sensitivity, and specificity for early detection and monitoring of cancer progression.
研究领域:
生物医学工程
·
癌症研究
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
Nanoscale
·
2020
·Vol. 12
·期 47
·24100-24106
·
IF 6.895
Xia Li,Fang Yang,Chunfang Gan,Ruo Yuan,Yun Xiang
The simultaneous imaging of the dynamic expression variations of regulatory RNAs in cells, which remains a major challenge, has important applications in precise disease diagnosis, treatment and prognosis. Here, we describe the establishment of a biodegradable ZnO nanoparticle (NP)-assisted asymmetric amplification approach for the simultaneous imaging of microRNA-21 (miRNA-21) and programmed cell death 4 (PDCD4) mRNA at distinct expression levels in live cells. The DNA signal probe complexes are immobilized on the ZnO NPs and readily delivered into the target cancer cells via the endocytosis pathway. The acidic microenvironment in cancer cells leads to the dissolution of the ZnO NPs to release Zn2+ ions and the intracellular miRNA-21 activates the Zn2+-dependent DNAzyme to cleave the substrate signal probes with the assistance of the Zn2+ cofactor to show green fluorescence for imaging miRNA-21. Meanwhile, the PDCD4 mRNA can displace the other quenched signal probes to generate red fluorescence. Importantly, the PDCD4 mRNA sequences can be recycled and reused by using the DNAzyme-cleaved sequences as the fuel strands through two strand displacement reactions to yield amplified red fluorescence for detecting low levels of PDCD4 mRNA. Moreover, our approach can be used to evaluate the varied expression levels of miRNA-21 and PDCD4 mRNA responsive to different drugs in cells, reflecting its usefulness for precise cancer diagnosis and prognosis upon anticancer drug treatment.
研究领域:
分子生物学
·
癌症研究
·
纳米医学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）