Analytical Methods
·
2026
·
IF 3
Jing Chen,Yanna Le,Luoxian Yang,Hua Qin,Wanli Yang
Chronic exposure to lead ions (Pb2+) disrupts intestinal flora homeostasis, underscoring the need for sensitive and practical detection methods. Herein, we report a dual-mode analytical strategy combining split-DNA re-assembly-driven colorimetric reaction with CRISPR/Cas12a trans-cleavage activity for Pb2+ analysis. A bifunctional magnetic probe is constructed, where Pb2+-specific DNAzyme recognition triggers cleavage and release of crRNA, which subsequently activates Cas12a to generate fluorescence by cleaving a fluorophore-quencher reporter. Concurrently, the remaining magnetic bead-anchored trigger initiates split G-quadruplex re-assembly, catalyzing a visible colorimetric reaction. Under optimized conditions, the fluorescence mode achieves a detection limit of 1.03 fM with a linear range of 5 fM to 100 pM, while the colorimetric mode offers a limit of 62.1 fM from 100 fM to 500 pM. Both modes exhibit excellent specificity against competing metal ions. The platform's clinical feasibility is validated using human serum samples, showing strong correlation with ICP-MS, satisfactory recoveries (97.1–103.4%), and good repeatability (CV = 4.1%). Magnetic separation effectively minimizes matrix interference, making the assay suitable for complex biological specimens such as intestinal contents and fecal samples. This dual-mode design integrates fluorescence for precise quantification and colorimetry for equipment-free visual detection, holding great promise for point-of-care testing, early risk assessment of lead-induced gut microbiota dysbiosis, and environmental monitoring.
✓ 已验证
DOI
PubMed
高影响因子
ACS Sensors
·
2026
·
IF 10.9
Dongmei Wang,Changjiang Ye,Tianshu Lan,Haobo Sun,Xueji Zhang,Yongxin Li
Exosomes have emerged as promising biomarkers for noninvasive cancer diagnosis, while highly sensitive, reliable, and convenient assays remain challenging, particularly with conventional single-signal approaches. Herein, we report an exosome-tailored dual-mode self-referencing biosensing platform on a single gold nanowire (AuNW) substrate for the accurate and sensitive quantification of HepG2-derived exosomes, integrating surface-enhanced Raman scattering (SERS) and electrochemical readouts on a AuNW substrate with dual-functional Au nanotag amplification. The platform features three exosome-specific design strategies: CD63 aptamer-functionalized DNA tetrahedrons (AD-NTH) with well-controlled probe orientation, spacing, and accessibility to enhance exosome capture efficiency and mitigate interfacial steric hindrance; dual recognition via CD63-mediated capture and AFP-mediated signal tagging for improved specificity toward HepG2-derived exosomes; and a ratiometric SERS/electrochemical dual-mode readout on a unified substrate to enhance signal reliability. This self-referencing mechanism effectively suppresses nonspecific interference and signal fluctuation, while the dual-functional Au nanotags provide amplified signal outputs. The platform achieves detection limits of 8.41 × 102 particles mL−1 (SERS) and 1.07 × 103 particles mL−1 (electrochemical). Clinical evaluation using serum samples from 10 liver cancer patients and 10 healthy volunteers shows statistically significant differentiation (P < 0.0001), supporting its promising application in noninvasive early screening and diagnosis of liver cancer.
✓ 已验证
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Ming Sui,Yanni Huang,Yang Tang,Jiaoyang Luo,Yiyang Dong,Yifei Guo,Yonggui Ma,Wei Gu,Mengyue Guo,Jingdan Huang,Dandan Kong,Jinping Li,Meihua Yang
Medicinal teas (MT) were popular in Asian countries for its various effective components. However, Pb pollution poses a serious threat to MT consumption due to the irreversible effect on enzyme activity and neurodegenerative disease. Owing to its toxicity and matrix interference, a rapid screening method for Pb at trace level with high sensitivity and accuracy is urgent for MT consumers. Herein, a simple and ultra-sensitive AuNPs/MWCNTs-MB/DNA-GCE quadruplex biosensor was developed and investigated. Specifically, the monitoring technology of scanning electrochemical microscope was utilized to analyze the assembly and application process for screening key parameters in complex matrix , through in-situ characterization of the microstructure changes on stepwise modified sensors. According to the combined results of electrochemical test and Randles-Sevcik equation, the proposed sensor showed a wide linear range from 5.0 * 10 −15 to 5.0 * 10 −13 M with LOD of 0.9 fM. In addition, the optimized sensor exhibited an excellent selectivity under the inference of metals and medicinal components. The proposed biosensor was successfully implemented for the ultrasensitive monitoring Pb pollution in MT while its water infusions recoveries maintained within acceptable range (81–118 %, 82–129 %). In all, AuNPs/MWCNTs-MB/DNA-GCE provided a new insight into the design, optimization, application and mechanism analysis of sensor for reliable sensing in controlling MT pollution and consumption safety.
✓ 已验证
产品关联
DOI
高影响因子
ANALYST
·
2022
·Vol. 147
·期 24
·5623-5632
·
IF 5.227
Jiahai Wang,Le Luo,Yunhui Li,Lanfang Chen,Cenlin Gui,Jianji Zhu,Huizhen Li,Wenlong Wang,Daqi Chen
Solid-state nanopores have been proven as a powerful platform for label-free single-molecule analysis. However, due to its relatively low resolution and selectivity, developing biosensors with good translocation signals faces two significant challenges: (1) small-sized chemical or biological targets show difficulty in producing recognizable translocation signals because of their weak interaction with the nanopore and (2) protein interferents that widely exist in biological samples or buffers would considerably deteriorate the noise level of the nanopore, submerging the translocation signal. Herein, we demonstrate an effective way to overcome both the challenges. DNA cubes were used as signal transducers that can achieve an ultra-high (>50 : 1) signal-to-noise ratio (SNR) translocation signal, which is maintained even in protein interferent-rich buffers. A sensing strategy was constructed via hepatitis B virus (HBV) target-triggered cleavage of the component elements of the DNA cube with the assistance of the CRISPR–Cas12a technology, which caused a great drop in the translocation rate. The elements to cleave were optimized, and the sensor performance was tested in different protein stabilizer-rich buffers and human serum. Coupling with the polymerase chain reaction (PCR) pre-amplification technology, HBV-positive or -negative classification was achieved with the detection limit reaching 5 aM. It is worth noting that in our method, all reaction buffers were directly used without further optimization, which is of great help for the practical application of solid-state nanopores.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Jieyu Hu,Xingxing Yu,Xiumei Zhuang,Yan Sun,Junjie Wang,Haiting Ren,Shenghao Zhang,Yueshou Zhang,Hongxing Qiu,Yongjun Hu
The p53 gene is a known cancer marker. We report a novel protocol for the SERS tandem strategy to detect the p53 gene with high sensitivity. Herein, the click reaction between azide and alkyne was catalyzed by utilizing copper oxide nanoparticles (CuONPs), which were enriched by a T-DNA-triggered hybridization chain reaction (HCR). The T-DNA signal was amplified by establishing the correlation between the T-DNA signal and the concentration of CuONPs in a nonenzymatic isothermal environment. In contrast to other Raman reporters, we used alkynyl compounds as Raman reporters, which showed excellent characteristics in the Raman-silent region (1800–2800 cm −1 ). Therefore, the highly sensitive and highly selective SERS signals could be obtained in complex biological matrices. Due to utilizing multistep amplification strategies, including the nanoparticle-modified HCR polymer and “click” reaction, the limit of detection (LOD) and the limit of quantification (LOQ) of this sensor could be as low as 0.0174 pM and 0.0583 pM, respectively. The accuracy of the strategy expressed as the RSD was in the range of 3.14%–6.21%. The results indicated that the constructed sensor has excellent performance for the detection of the p53 gene in serum samples in a low concentration range, which suggests that the proposed enzyme-free SERS analytical sensor has good clinical application prospects.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2016
·Vol. 6
·期 42
·35952-35965
·
IF 3.289
Qian Gan,Chun-Lian Zhang,Bing-Feng Wang,Ya-Hong Xiong,Yin-Lian Fu,Zong-Wan Mao,Xue-Yi Le
Two novel mononuclear mixed ligand copper(II) complexes, [Cu(Gly-L-val)(HPB)(H2O)]·ClO4·1.5H2O (1) and [Cu(Gly-L-val)(PBT)(H2O)]·ClO4 (2) (Gly-L-val = glycyl-L-valine, HPB = 2-(2′-pyridyl)benzimidazole, PBT = 2-(2′-pyridyl)benzothiazole), have been synthesized and characterized using various analytical and spectroscopic methods. The interactions of the complexes with DNA have been explored by viscometry, thermal denaturation, cyclic voltammetry (CV), agarose gel electrophoresis and spectroscopic means (UV absorption, circular dichroism (CD) and fluorimetry), as well as molecular docking techniques. These studies confirmed the mode of the complexes bound to calf thymus DNA (CT-DNA) through insertion with certain affinity (Kb = 3.211 × 105 M−1 for 1 and 4.734 × 104 M−1 for 2). In the fluorimetric experiments of thermodynamics (KSV = 1.145 × 104 M−1 for 1 and 2.634 × 103 M−1 for 2), the changes in enthalpy (ΔH > 0), entropy (ΔS > 0) and Gibbs free energy (ΔG < 0) in the interactions between the complexes with DNA suggested that the process occurred spontaneously through hydrophobic interactions. The complexes displayed oxidative cleavage of pBR322 plasmid DNA in the presence of ascorbic acid, probably induced by ˙OH as a reactive oxygen species. Furthermore, the molecular docking technique was applied to ascertain the mode of action for the complexes towards DNA. Moreover, superoxide dismutase (SOD) activity studies were performed using the photoreduction of nitroblue tetrazolium (NBT) under a non-enzymatic system and the antioxidant activities of 1 and 2 determined with IC50 values of 0.337 and 0.146 μM, respectively. The cytotoxicity of the Cu(II) complexes against A549, HeLa, PC-3 tumor cell lines and NIH3T3 (non-tumor cell line) was studied by an MTT assay and it was found that 1 exhibited better cytotoxicity against A549 and PC-3 than 2 and the widely used drug cisplatin.
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2023
·
IF 13.6
Xiaoying Wang,Mingming Xu,Yijing Kuang,Xiangping Liu,Jinhua Yuan
Simultaneous quantification of multiple heavy metal ions (HMIs) is essential due to enhanced toxicity induced by synergistic effects. The currently available detection methods suffer from drawbacks such as costly devices, poor anti-interference, and specificity. In this work, a ratiometric electrochemical aptasensor for simultaneous detection of trace lead (Pb 2+ ) and mercury ions (Hg 2+ ) was developed. Here, a metal-organic framework, UiO-66-CNTs, with inherent and stable electrochemical signal was used for loading complementary strands (CSs) and internal reference. Guanine-rich and thymine-rich oligonucleotides , labelled with carbon dots (CDs), acted as aptamers (Apts) and hybridized with CSs to form M-shaped DNA complexes. Pb 2+ and Hg 2+ could be recognized and captured by Apts to form Pb 2+ -G-quadruplex and T-Hg 2+ -T complexes, leading to the destruction of M-shaped DNA complexes and changes in CDs’ signal. The current ratios, I CDs / I UiO-66-CNTs , were applied to quantify Pb 2+ and Hg 2+ . Benefiting from the anti-interference ability of the ratiometric strategy and the specificity of Apts, the proposed method enabled detection Pb 2+ and Hg 2+ through simple instrumentation, with detection limits of 2.0 ng mL −1 and 0.5 ng mL −1 . Moreover, it was applied to assess Pb 2+ and Hg 2+ contamination in series of aquatic edible vegetables. The experimental results were consistent with inductively coupled plasma-mass spectrometry (ICP-MS), demonstrating its potential in practical applications.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Nuanfei Zhu,Xuesong Li,Ye Liu,Jingfu Liu,Yawei Wang,Xiangyang Wu,Zhen Zhang
Dibutyl phthalate (DBP) is considered as one of the most widely used phthalate esters (PAEs), which has attracted worldwide concerns because of its potential threats to eco-environments and human health. Systematic investigations of DBP environmental occurrence contribute to the further risk assessment, which depends on effective and available analytical methods. In this study, an amplified ratiometric fluorescence ELISA was established for sensitive and high-throughput detection of DBP in the aquatic system based on a novel tetrahedral DNA nanostructure (TDN)-scaffolded-DNAzyme (Tetrazyme). Wherein, Tetrazyme was prepared by the precise folding of G-quadruplex sequence on three vertex angles of the TDN, together with hemin as the horseradish peroxidase (HRP)-mimicking enzyme. The rigid TDN avoided the local overcrowding effect to provide a reasonable spatial spacing on the interface for G-quadruplex sequence, increasing the collision chance between DNAzyme and substrates, improving the catalytic ability of DNAzyme effectively. Besides, streptavidin (SA) and biotin (bio) were used to anchor TDN and antibody, in which the specific binding of SA/bio could make more Tetrazyme conjugate on each signal element, resulting in the dual signal amplification. Meanwhile, the accuracy and precision were enhanced owing to the inherent built-in rectification to the environment from the dual output ratiometric fluorescence assay. Under the optimized conditions, the detection limit of this proposed method was 0.17 ng/mL (16 times lower than that of conventional ELISA using the same antibody) with a satisfactory accuracy (recoveries, 79.0%- 116.2%; CV, 2.1–6.5%). Overall, this platform provides a promising way for accurate, sensitive and rapid determination of DBP from environmental waters.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2020
·
IF 3.004
Ying Sun,Li Xie,Fulin Feng,Qing Han,Lanlan Wei,Zigang Tang,Xuejun Kang
The determination of the concentrations of urinary biomarkers of oxidative damage to DNA and RNA is difficult due to the low content of targets and the complex matrix of urine. A method using polystyrene/polypyrrole (PS/PPY) electronspun nanofibers as the adsorbent was introduced to the routine urinary treatment and determination of 8-OHdG and 8-oxoG for the first time. And 2-aminoethyl diphenylborate (DPBA) solution was creatively used in the loading and rinsing steps in order to promote the retention of the analytes as well as remove impurities. Under optimal conditions, 8-OHdG, 8-oxoG and IS were separated very well and exhibited a good linearity in the range of 0.5–50 ng mL −1 , with correlation coefficients of R 2 > 0.996. Limits of detection (LOD) were 0.058 ng mL −1 and 0.093 ng mL −1 , and limits of quantification (LOQ) were 0.195 ng mL −1 and 0.309 ng mL −1 , respectively. The recoveries were 88.8–104.9%. The proposed method was so simple and economical that it had the potential to be applied to batch quantitative analysis of 8-OHdG and 8-oxoG in urine. And it was successfully applied to real urine samples of cancer patients.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL LETTERS
·
2021
·
IF 2.329
Penghui Jin,Daoqing Ma,Yabiao Gao,Libin Wang,Zhihong Gao,Yating Zhang,Mingxin Liu,Jingying Xu,Jidong Wang
A highly sensitive and dual-directional fluorescent sensing system was developed to detect both cisplatin prodrug cross-linked hyaluronic acid (CPHA) hydrogel (quencher) and DNA (receptor) based on mercaptopropionic acid (MPA) capped CdTe quantum dots (QDs). Different from the previous work for this platinum compound, a multifunctional delivery system, CPHA hydrogel, which was used to address the challenges in drug resistance and severe side effects, was prepared and accurately detected based on the synthesis of the cisplatin prodrug and fluorescence quenching of CdTe QDs resulting from photo-induced electron transfer. When DNA was conjugated with Pt ions in a system composed of CdTe QDs and CPHA hydrogel, the fluorescence of the system was recovered for the determination of DNA. This reversible “turn off–on” strategy presented a high sensitivity and low detection for CPHA hydrogel from 10 to 50 nM with a detection limit of 1.50 nM and DNA from 0 to 75 nM with a detection limit of 0.60 nM. This approach provides a novel direction for further study of the Pt based drug-load with reduced side effects and improved therapy efficacy, effective screening of anticancer drugs, and accurate determination of DNA at low concentrations.
✓ 已验证
产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-8
·
IF 5.479
Song Xiaolei,Wang Yu,Liu Su,Zhang Xue,Wang Jingfeng,Wang Haiwang,Zhang Fenfen,Yu Jinghua,Huang Jiadong
An amplified electrochemical biosensing scheme is described for lead(II) ions. It is making use of DNAzyme-assisted target recycling and catalytic hairpin assembly (CHA). The hairpin strand (substrate probe for the Pb 2+ -based DNAzyme; referred to as SP) is composed of trigger probe (TP) and a capture probe 1 attached to gold nanoparticles (AuNP). In the presence of the enzyme probe that partially hybridizes with SP, the introduction of Pb 2+ triggers target recycling and drives the highly amplified translation of target Pb(II) to TP. The CHA reaction is further initiated by TP. The modified AuNP act as the connecting unit, and this leads to the formation of a 3D DNA-AuNP network on the electrode (which is the third amplification step). It can bind the positively charged redox mediator RuHex via electrostatic interaction for electrochemical detection. This biosensor has a low detection limit (95 pM) and any analytical range that covers the 100 pM to 5 μM Pb(II) concentration range. It is selective over other divalent metal ions. It was applied to the determination of Pb 2+ in spiked real-world samples.
Schematic presentation of the electrochemical biosensor. The triply amplified electrochemical assay is based on the use of DNAzyme-assisted target recycling with catalytic hairpin assembly (CHA) reaction for sensitive and selective determination of lead ion (Pb 2+ ). AuNP: gold nanoparticles; SP: substrate probe; EP: enzyme probe.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 9
·1-9
·
IF 5.705
Wang Jing,Li Haigang,Li Tingting,Ling Liansheng
A one-step, one-tube colorimetric assay is described for the detection of bacterial double-stranded DNA (dsDNA). It utilizes a G-quadruplex DNAzyme produced by an asymmetric polymerase chain reaction (As-PCR) that catalyzes the oxidation of cysteine to form cystine. This results in the formation of oligonucleotide-modified gold nanoparticles via triplex formation, and eventually in a color change from red to blue that occurs within about 10 mins. This can be measured by ratiometric colorimetric (at 525 and 600 nm). The limit of detection (LOD) for the model analyte (dsDNA of Staphylococcus aureus ( S. aureus )) is as low as 0.28 pg per 0.05 mL with a good linear response ranging from 16.0 fg·μL −1 to 1.6 ng·μL −1 . This is much lower than previously reported LODs. The assay is highly selective for S. aureus dsDNA over a range of other bacterial DNAs. Conceivably, it provides an attractive alternative tool for rapid detection of bacterial dsDNA as required in pathogen screening in the food industry.
Schematic presentation of a colorimetric assay for bacterial DNA. It is based on the catalytic activity of a G-quadruplex DNAzyme that is formed by an asymmetric PCR involving triplex DNA formation and gold nanoparticle (AuNPs) aggregation.
✓ 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）