Biosensors-Basel
·
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
·
分析化学
·
纳米技术
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2023
·Vol. 16
·期 1
·51-61
·
IF 3.1
Jing Qi,Qianyi Qi,Zhou Zhou,Yixuan Wu,Aiting Cai,Jinran Wu,Bairong Chen,Qingxiang Wang,Lin Chen,Feng Wang
The detection of epidermal growth factor receptor (EGFR) mutation L858R in circulating tumor DNA (ctDNA) is beneficial for the clinical diagnosis and personalized therapy of non-small cell lung cancer (NSCLC). Herein, for the first time, the combination of the primer exchange reaction (PER) and clustered regularly interspaced short palindromic repeats (CRISPR) and its associated nucleases (Cas) 14a was used in electrochemical biosensor construction for the detection of ctDNA EGFR L858R. EGFR L858R, as the target, induced the isothermal amplification of the PER reaction, and then the CRISPR/Cas14a system was activated; subsequently, the substrate ssDNA-MB was cleaved and the electron on the surface of the gold electrode transferred, resulting in the fluctuation of the electrochemical redox signal on the electrode surface, whereas the electrochemical signal will be stable when EGFR L858R is absent. Therefore, the concentration of EGFR L858R can be quantified by electrochemical signal analysis. The low detection limit is 0.34 fM and the dynamic detection range is from 1 fM to 1 μM in this work. The PER-CRISPR/Cas14a electrochemical biosensor greatly improved the analytical sensitivity. In addition, this platform also exhibited excellent specificity, reproducibility, stability and good recovery. This study provides an efficient and novel strategy for the detection of ctDNA EGFR L858R, which has great potential for application in the diagnosis and treatment of NSCLC.
研究领域:
分子生物学
·
临床诊断
·
癌症研究
·
生物传感技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Yunyun Hu,Xin Guo,Peilin Gu,Qin Luo,Yang Song,Erqun Song
Since circulating tumor DNA (ctDNA) has been regarded as a promising noninvasive biomarker for early cancer screening, cancer condition monitoring, and prognosis evaluation, the accurate and simple detection of ctDNA has been extensive explored. Up to now, there is still a lot of space for following-up the assay of ctDNA, especially simply direct analysis ctDNA in complex biological samples such as blood sample. In this work, we develop a Mn 2+ -mediated magnetic relaxation switching (MRS) assay for direct and simple detection of ctDNA via exonuclease III-assisted signal amplification. In the presence of target ctDNA, exonuclease III selectively cleaves the hybridization complex of between ctDNA and its complementary DNA labeled with magnetic beads and alkaline phosphatase (MB 1000 -DNA 1 -ALP) to release ALP and ctDNA, the former further generating Mn 2+ and causing transverse relaxation time signal change while the later recycling to amplify the signal. The combination of efficient recognition of ctDNA, enzyme-assisted amplification, and the MRS assay could achieve direct and specific detection of ctDNA with a detection limit of 340 pM. In our opinion, this is the first successful direct detection of ctDNA in whole blood sample without ctDNA extraction from biospecimen, which make the assay for ctDNA much accurate and timely since a short half-life (2 h) of ctDNA in blood. The as-proposed method for ctDNA assay in this study shows potential tracking the evolution of tumor timely and serving as a liquid biopsy for diagnosis.
研究领域:
生物医学工程
·
分子诊断学
·
癌症研究
✓ 已验证
产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 9
·1-9
·
IF 5.3
Song Yiwei,Jin Xiuyan,Zhao Yiou,Cheng Shuwen,Xu Sai,Bu Shengjun,Liu Liming,Zhou Chunyang,Pang Chunying
Single-level biomarker detection has the limitation of insufficient accuracy in cancer diagnosis. Therefore, the strategy of developing highly sensitive, multi-channel biosensors for high-throughput ctDNA determination is critical to improve the accuracy of early diagnosis of clinical tumors. Herein, in order to achieve efficient detection of up to ten targets for early diagnosis of ovarian cancer, a DNA-nanoswitch-based multi-channel (DNA-NSMC) biosensor was built based on the multi-module catalytic hairpin assembly-mediated signal amplification (CHA) and toehold-mediated DNA strand displacement (TDSD) reaction. Only two different fluorescence signals were used as outputs, combined with modular segmentation strategy of DNA-nanoswitch-based reaction platform; the multi-channel detection of up to ten targets was successfully achieved for the first time. The experimental results suggest that the proposed biosensor is a promising tool for simultaneously detecting multiple biomarkers for the early diagnosis of ovarian cancer, offering new strategies for the early screening, diagnosis, and treatment not only for ovarian cancer but also for other cancers. Graphical abstract
研究领域:
分子生物学
·
医学诊断
·
生物传感技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 23
·5770
·
IF 4.2
Lili Fu,Kun Liu,Jinyu Yang,Yuan Zhao,Zhijun Wang,Dongxu Tang,Yuesheng Li,Huangqin Chen
In this study, a polyvinyl alcohol/polyethylene glycol/hydroxypropyltrimethyl ammonium chloride chitosan (PVA/PEG/HACC) ternary composite hydrogel was synthesized using electron-beam radiation. The materials were thoroughly characterized via Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, thermogravimetric analysis, Brunauer–Emmett–Teller analysis, gelation fraction tests, and swelling rate tests. Systematic adsorption experiments revealed that the rate of adsorption of calf thymus DNA (ctDNA) by the PVA/PEG/HACC hydrogel reached 89%. The adsorption process followed the Langmuir isotherm and pseudo-second-order kinetic model. This process was mainly characterized by monolayer chemical adsorption, with intraparticle diffusion playing a crucial role. In addition, the process was spontaneous, with higher temperatures enhancing adsorption. The possible adsorption mechanisms included electrostatic interactions, hydrogen bonding, and van der Waals forces. The maximum ctDNA desorption rate was 81.67%. The adsorption rate remained at 71.39% after five adsorption–desorption cycles. The bioactivity of the PVA/PEG/HACC hydrogel was validated by antibacterial, cytotoxicity, and apoptosis tests. The results of this study demonstrated the crucial application potential of adsorbent materials in DNA adsorption and biomedical applications.
研究领域:
材料科学
·
生物医学工程
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Nano
·
2024
·
IF 15.8
Zhiying Wang,Ming Liu,Haocheng Lin,Guiying Zhu,Zaizai Dong,Nan Wu,Yubo Fan,Gaolian Xu,Lingqian Chang,Yang Wang
Strategies for rapid, effective nucleic acid processing hold tremendous significance to the clinical analysis of circulating tumor DNA (ctDNA), a family of important markers indicating tumorigenesis and metastasis. However, traditional techniques remain challenging to achieve efficient DNA enrichment, further bringing about complicated operation and limited detection sensitivity. Here, we developed an ion concentration polarization microplatform that enabled highly rapid, efficient enrichment and purification of ctDNA from a variety of clinical samples, including serum, urine, and feces. The platform demonstrated efficiently separating and enriching ctDNA within 30 s, with a 100-fold improvement over traditional methods. Integrating an on-chip isothermal amplification module, the platform further achieved 100-fold enhanced sensitivity in ctDNA detection, which significantly eliminated false-negative results in the serum or urine samples due to the low abundance of ctDNA. Such a simple-designed platform offers a user-friendly yet powerful diagnosis technique with a wide applicability, ranging from early tumor diagnosis to infection screening.
研究领域:
生物医学工程
·
分子生物学
·
临床诊断
·
癌症研究
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Xiao Yang,Liangyi Zhao,Siyi Yang,Miao Tang,Huanbao Fa,Danqun Huo,Changjun Hou,Mei Yang
The detection of circulating tumor DNA (ctDNA), as a practical liquid biopsy technique, was of great significance for the study of cancer diagnosis and prognosis. However, reported methods for detection ctDNA still have some limitations, such as tedious process and high cost. In this study, CsPbBr 3 nanosheet (CsPbBr 3 NS) with high water stability was prepared by etching, and its fluorescence intensity could be stably stored for 1 year. The Ti 3 C 2 T x possessed high quenching efficiency for CsPbBr 3 NS and the HOMO-LUMO orbital study revealed that the PET mechanism was responsible for fluorescence quenching. And the Ti 3 C 2 T x showed stronger affinity towards single-stranded DNA (ssDNA), as compared with double-stranded DNA (dsDNA). The probe ssDNA could be adsorbed on the surface of Ti 3 C 2 T x through π-π stacking. After the targets were recognized by probe ssDNA to form dsDNA, its affinity with Ti 3 C 2 T x decreased and the active site of Ti 3 C 2 T x recovered, causing a high quenching efficiency on CsPbBr 3 NS. Based on this, a label-free fluorescent biosensor was designed for the sensitive detection of ctDNA ( EGFR 19 Dels for non-small cell lung cancer, NSCLC). Under the optimal experimental conditions, this biosensor exhibited a detection limit of 180 fM and a linear range of 50 pM–350 pM with amplification of magnetic beads through strand displacement reaction. In addition, this sensor was applied to the detection of ctDNA in serum samples and cells lysates. This method for ctDNA detection was expected to have great potential for biomarker detection in the field of liquid biopsy.
研究领域:
癌症诊断
·
纳米技术
·
生物传感器
·
分子生物学
·
生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·
IF 4.2
Xiao Yang,Yang Huang,Siyi Yang,Miao Tang,Juan Liu,Jinhui Shen,Huanbao Fa,Danqun Huo,Changjun Hou,Mei Yang
Technological advances in the detection of circulating tumor DNA (ctDNA) have made new options available for diagnosis, classification, biological studies, and treatment selection. However, effective and practical methods for analyzing this emerging class of biomarkers are still lacking. In this work, a fluorescent biosensor was designed for the label-free detection of ctDNA (EGFR 19 del for non-small cell lung cancer, NSCLC). The biosensor was based on the fact that MnO2 nanosheets (MnO2 NSs) have stronger affinity towards single-stranded DNA (ssDNA), as compared with double-stranded DNA (dsDNA). As a high-performance nanoenzyme, MnO2 NSs could oxidize dopamine (DA) into fluorescent polydopamine nanoparticles (FL-PDA NPs), which could be used as a fluorescence signal. The probe ssDNA could be adsorbed on the surface of MnO2 NSs through π–π stacking, and the active site would be masked, causing a lower fluorescence. After the targets were recognized by probe ssDNA to form dsDNA, its affinity for MnO2 NSs decreased and the active site recovered, causing a restored fluorescence. It was verified that Mn ions, •OH radicals and electron transfer were the important factors in the catalytic oxidation of DA. Under the optimal experimental conditions, this biosensor exhibited a detection limit of 380 pM and a linear range of 25–125 nM, providing reliable readout in a short time (45 min). This sensor exhibited outstanding specificity, stability and reproducibility. In addition, this sensor was applied to the detection of ctDNA in serum samples and cell lysates. It is demonstrated that FL-PDA NPs can be used as a fluorescence signal for easy, rapid and label-free detection of ctDNA without any other amplification strategies, and the proposed strategy has great potential for biomarker detection in the field of liquid biopsy.
研究领域:
生物技术
·
生物传感器
·
纳米技术
·
癌症诊断
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Lei Guo,Zhaode Mu,Bin Yan,Jie Wang,Jing Zhou,Lijuan Bai
Non-small cell lung cancer (NSCLC) is the most common type of lung cancer with poor survival, in part because that half of the cases are already in advanced stage when they are diagnosed. In response to this issue, the present work constructed a novel electrochemical biosensor for sensitive detection of circulating tumor DNA (ctDNA) related to NSCLC. Specifically, polyethyleneimine (PEI) and covalent organic framework (COF TAPB-TFPB ) functionalized nitrogen-doped graphene (NG) nanocomposite (NG-PEI-COF TAPB-TFPB ) was designed and employed as sensing platform for the first time. NG-PEI-COF TAPB-TFPB with porous structure and good conductivity showed a synergistic effect on promoting electron transfer, which could provide large specific surface area and active sites to increase the immobilization of capture probe (CP). In addition, gold nanoparticles (AuNPs) decorated Fe-based metal-organic framework (Fe-MOF) were labeled with signal probe (SP) to form tracer label, which would attach to the electrode surface by sandwich hybridization of target probe (TP) with CP and SP. Consequently, Fe III from Fe-MOF could react with K 4 [Fe(CN) 6 ] to form electroactive prussian blue, generating a significantly amplified electrochemical detection signal. Under optimum conditions, the proposed DNA biosensor displayed a broad linear range for ctDNA determination from 100 fM to 100 nM with a detection limit of 7.65 fM. Moreover, the biosensor was successfully fabricated to assay ctDNA in human serum, indicating the potential for cancer diagnosis. More importantly, the high sensitivity and satisfactory stability of this biosensor make it a promising method to detect other biomolecules by changing the corresponding sequences of CP and SP.
研究领域:
生物医学工程
·
癌症生物学
·
分析化学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Lijia Yuan,Ting Xiao,Jing Li,Weifeng Duan,Duojie Bianba,Ligai Zhang,Yuan He,Dunquan Xu,Muzhen Da,Jing Sheng,Ming Chen,Sha Yang
Real-time circulating tumor DNA (ctDNA) monitoring allows early detection of residual disease, dynamic assessment of treatment response, and identification of resistance, highlighting the demand for scalable point-of-care tests that allow mutations to be resolved. Here, we introduce MED, a multiplexed, entropy-driven catalysis (EDC), single-nucleotide-resolved ctDNA detection assay, enabling enzyme-free nucleic-acid amplification and visual detection of ctDNAs. The recognition of ctDNA depends on the initiation of EDC and magnetic separation with single-nucleotide resolution. The assay leverages a triple amplification strategy to amplify the recognition events of trace ctDNAs, involving EDC, Ag+ release from silver nanoparticles (AgNPs) catalyzed by H2O2, and urea hydrolysis catalyzed by urease, for colorimetric readout of pH changes. For 40 blood specimens (20 colorectal cancer, 10 benign colorectal adenoma, and 10 normal controls), the assay simultaneously detected the presence of a panel of four kinds of ctDNAs in 100% concordance with sequencing and demonstrated over 85% accuracy in distinguishing between cancer and noncancer. Further, integrated into bioactive paper and smartphone readout, MED provides rapid (sample-to-answer time within 45 min), multiplexed, and accessible point-of-care results, with over 70% accuracy. Its unique capacity surpasses existing detection platforms, presenting robust clinical potential to enhance early diagnosis, enable dynamic monitoring of cancer progression, and guide personalized therapeutic interventions.
研究领域:
分子诊断
·
癌症基因组学
·
生物传感器
·
即时检测
·
核酸检测
·
临床化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS
·
2022
·
IF 1.449
Xin-Yan Lu,Yan-Yue Lou,Kai-Li Zhou,Shao-Liang Jiang,Jie-Hua Shi
In this paper, the interacting characteristics of febuxostat (FBST), an inhibitor of xanthine oxidase for treating gout patients with hyperuricemia with calf thymus DNA (ctDNA) was investigated through multi-spectroscopic methodologies combined with theoretical calculation for understanding the interacting mode on ctDNA, affinity with ctDNA, interacting forces, as well as the alteration in the conformation of ctDNA after interacting FBST The experimental results demonstrated that interacting FBST with ctDNA formed 1:1 complex, the association constant was 913 M−1 at 298 K, suggesting the affinity of FBST on ctDNA was very weak, the interacting mode of FBST on ctDNA was groove binding, and it inserted into the minor groove with rich A-T region of ctDNA. Based on the results of the thermodynamic analysis and theoretical calculation, it can be inferred that the dominated interacting forces between FBST and ctDNA were van der Waals forces and hydrogen bond. And, interacting FBST with ctDNA was a spontaneous, enthalpy-driven, and exothermic process because of ΔG0 < 0, ΔH0 < 0, and |ΔH0| > T|ΔS0|. The results of the circular dichroism (CD) measurements indicated the conformation of ctDNA was weakly disturbed after interacting with FBST but still maintained B-conform. The studied results offer significant insight into further clarifying whether it has genotoxicity.
研究领域:
生物化学
·
药物化学
·
分子生物学
✓ 已验证
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Jiangli Song,Hanlian Chen,Rongkai Pan,Fangyu Jin,Jie Tang,Qianqian Huang,Guohua Zhou,Xiumei Song
Understanding the interactions between small molecules and calf thymus deoxyribonucleic acid (ctDNA) is critical for certain aspects of drug discovery. In this study, three 11 H -indeno[1,2-b]quinoxalin-11-one thiosemicarbazones were synthesized and their interaction with ctDNA was examined through various spectroscopic techniques, including ultraviolet (UV) spectroscopy, fluorescence spectroscopy, and circular dichroism (CD) spectrum, and through physicochemical methods, including viscosity measurements. In addition, the effects of these thiosemicarbazone compounds 4a, 4b and 4c on several cancer cell lines were explored. The results of UV absorption, fluorescence quenching and CD experiments indicated that compounds 4a, 4b and 4c primarily bound to ctDNA by an intercalation. This mode of interaction was further corroborated by viscosity measurements. Docked poses of compound 4a revealed that it formed a crucial N position hydrogen bond with the DNA receptor. The chemical structure of compound 4a was further confirmed by X-ray crystallographic analysis. Through biological evaluation, it was found that the in vitro cytotoxicity of three compounds agreed with the order of their binding strength to ctDNA, indicating that the antitumor activity of these compounds correlated with their binding affinity to ctDNA.
研究领域:
生物化学
·
药物化学
·
肿瘤学
✓ 已验证
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）