Scientific Reports
·
2017
·Vol. 7
·期 1
·1-7
·
IF 4.259
Cao Hongmei,Fang Xueen,Liu Peng,Li Hua,Chen Weiwei,Liu Baohong,Kong Jilie
Determination of proteins, especially low-abundance proteins with high sensitivity and specificity, is essential for characterizing proteomes and studying their biochemical functions. Herein, a novel Magnetic-Immuno-Loop-Mediated Isothermal Amplification (Im-LAMP) based on DNA-encapsulating liposomes (liposome-Im- LAMP), was developed for trace amounts of proteins. To the best of our knowledge, this is our first report about the magnetic Im-LAMP approach based on liposomes encapsulated template DNA as the detection reagent. The DNA template was released from liposomes and then initiated an Im-LAMP reaction, generating the fluorescence signal with high sensitivity and rapidity. This technique was applied for the determination of P-glycoprotein as a model protein. It was demonstrated that the technique exhibited a dynamic response to P-glycoprotein ranging from 1.6*10 −2 to 160 pg/ml with a greatly low detection limit of 5*10 −3 pg/ml (5 fg/ml) which is substantially better than conventional enzyme-linked immunosorbent assays (ELISA). This ultra sensitivity was attributed to the LAMP reaction initiated by the enormous DNA targets encapsulated in liposomes. This magnetic liposome-Im–LAMP as an alternative approach is attractive for applications in other low-abundance proteins detection in clinical diagnostics.
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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临床诊断
✓ 已验证
DOI
PubMed
高影响因子
RSC Advances
·
2022
·Vol. 12
·期 49
·31869-31877
·
IF 4.036
Yue Zheng,Shengquan He,Penghui Jin,Yabiao Gao,Ya Di,Liming Gao,Jidong Wang
Multifunctional therapeutic platforms with targeted delivery, fast diagnosis, and efficient therapy could effectively reduce side effects and improve treatment in the clinical therapy of tumors. Near-infrared DNA-templated CdTeSe quantum dots (DNA-CdTeSe QDs) were developed as building blocks to construct a multifunctional carboxymethyl cellulose (CMC)-based nanohydrogel as a nanocarrier to address the challenges of serious side effects and precise treatment in cancer theranostics, including active tumor targeting, fluorescence tracking, controlled drug release, chemotherapy and gene regulation. Single-stranded DNA containing the complementarity sequence of miRNA and cystine, as co-crosslinkers, initiated hybridization between the DNA-CdTeSe QD-modified CMC chain with the anti-nucleolin aptamer DNA (AS1411)-modified CMC chain to form the hydrogels. DOX, as a model drug, was successfully incorporated into the hydrogels. The synthesized multifunctional hydrogel nanocarriers with an average diameter of 150 nm could be taken up through targeting and achieved the controlled release of DOX by triggering both glutathione (GSH) and miRNA in the tumor microenvironment. The CdTeSe QDs trapped in nanohydrogels acted as fluorophores for bioimaging in the diagnosis and treatment process. The proposed multifunctional delivery system provided a potential platform for tumor imaging and precise therapy.
研究领域:
生物医学工程
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纳米医学
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肿瘤学
✓ 已验证
DOI
高影响因子
Analytical Methods
·
2025
·
IF 2.6
Jinling Cui,Ritong Sun,Yu Liao,Yanling Wang,Chao Jiang,Lei Wang,Chao Shi,Cuiping Ma
Electrochemical DNA biosensors have been extensively used in food safety, clinical medicine and environmental monitoring due to their high specificity and sensitivity. However, electrochemical DNA biosensors based on nucleic acid hybridization still face challenges in achieving rapid and sensitive detection. In this study, a sensitive and rapid electrochemical DNA biosensor was developed using Strand Exchange Amplification (SEA) technology, with its performance evaluated against the bovine genome as the target. Additionally, gold nanoparticles (AuNPs) were employed to modify the electrode surface, a strategy to enhance both the density of probe modification and the amplification efficiency. Furthermore, the biosensor's sensitivity has been shown to be augmented by the exceptional conductivity of AuNPs. Despite the biosensor's simplicity and sensitivity, the detection time remains a limiting factor. To address this, the incorporation of an electric field within the biosensor framework has been proposed as a strategy to enhance the coupling rate of the nucleic acid amplification and streptavidin–biotin systems. This modification is anticipated to reduce the overall detection time, enabling rapid and precise real-time nucleic acid analysis. The biosensor demonstrated the capability to detect genome DNA as low as 1 fg μL−1 within 65 min, underscoring its significant potential for applications, such as detecting meat adulteration.
研究领域:
生物传感技术
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分析化学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2017
·Vol. 41
·期 15
·7613-7620
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IF 3.269
Qianqian Pei,Yu Wang,Su Liu,Yifei Qin,Xueqi Leng,Xuejun Cui,Jiadong Huang
In this study, a novel electrochemical biosensor based on exonuclease III (Exo III)-aided autonomous cascade signal amplification for ultrasensitive and highly specific detection of Salmonella typhimurium (S. typhimurium) has been reported. Herein, two duplex DNA probes were ingeniously designed. An Ap–Pr duplex probe was constructed as a recognition element by hybridizing an S. typhimurium aptamer (Ap) with a primer (Pr). A P–S duplex probe has been designed to contain a protruding DNA fragment at 5′-termini that has an identical sequence with Pr (the secondary Pr), a complementary duplex DNA that is used for blocking a signal-triggered probe (S), and an overhung nucleic acid segment at 3′-end that can anneal with Pr and form double-stranded DNA with a blunt 3′-terminus, thus initiating Exo III-aided amplification reaction. Upon sensing of the target S. typhimurium, the specific recognition of S. typhimurium and aptamer releases Pr, which anneals with the P–S duplex and triggers the Exo III cleavage reaction, accompanied by release of Pr, S, and secondary Pr. After this, the released Pr and secondary Pr anneal with other P–S duplex, which in turn triggers the Exo III-assisted cycling cleavage. Thus, large amounts of S are liberated, which hybrids with S* (DNA probe complementary to S); thereby, 5′-methylene blue (MB)-labelled DNA probe (MBP) folds into a hairpin structure with 5′-terminus close to electrode surface for efficient electronic transfer. Moreover, the S–S* hybrid duplex is specifically hydrolyzed by Exo III to liberate S, which accordingly results in the cycling cleavage of S*. Therefore, numerous MBP transform into a hairpin structure with electroactive MB close to the electrode surface; this generates significantly amplified electrochemical signal towards S. typhimurium detection. The present developed strategy exhibited high sensitivity towards S. typhimurium with a detection limit of 2.8 × 10 cfu mL−1. Furthermore, the proposed autonomous cascade signal amplification strategy can be easily extended for detection towards a wide spectrum of analytes including nucleic acid, proteins, and small molecules possessing their corresponding aptamers. Therefore, the developed Exo III-aided autonomous cascade signal amplification strategy might indeed provide a facile and universal tool for bioanalysis and clinical diagnosis.
研究领域:
生物传感技术
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分子诊断
·
分析化学
✓ 已验证
DOI
高影响因子
Separation Science Plus
·
2018
·Vol. 1
·期 3
·217-224
·
IF 0
Lei Chen,Lei Wang,Dan Song,Zhangrun Xu
A three-dimensional reduced graphene oxide porous aerogel with packaged TiO 2 nanoparticles was prepared by a simple one-step self-assembly in a water bath at a mild temperature. Herein, we overcame the aggregation of graphene layers by forming three-dimensional architecture, and utilized TiO 2 aggregates as blocks to make the composite surface uneven. The extended aromatic areas contribute to DNA adsorption by aromatic stacking and hydrophobic interaction. The obtained composite exhibited porous structure and was positively charged at a low pH. The composite exhibited a high capacity of 890.1 mg g −1 and an adsorption efficiency of 98.4% to DNA at pH 3, however, bovine serum albumin at the same concentration level was hardly adsorbed. The composite showed favorable selectivity to DNA against protein even designed in such a simple manner. The retained DNA could be stripped by using a 0.04 mol L −1 Britton–Robinson buffer at pH 10, giving rise to a recovery of 96.3%. The DNA was isolated from human whole blood to be used as templates for PCR amplification successfully. This illustrates that DNA with high purity and quality was obtained.
研究领域:
材料科学
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生物技术
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医学检测
✓ 已验证
DOI
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Guoli Chen,Ji-Na Hao,Yongsheng Li
5-Methylcytosine (5-mC), as one of the primary forms of DNA methylation, plays essential roles in a variety of biological activities. Its rapid and precise detection is therefore paramount for abnormal DNA methylation assessment, but still remains challenging. Herein, a highly luminescent and stable lanthanide-based metal-organic framework (MOF) probe, namely Eu-ZrMOF, is firstly designed for qualitative and quantitative analysis of 5-mC, wherein the organic building units of 1,3,5-benzenetricarboxylic acid (BTC) serve as both the energy donors and recognition molecules, and the modified Eu 3+ ions function as intense signal reporters. Significantly, the luminescence of the fabricated Eu-ZrMOF probe can be remarkably quenched by 5-mC via its inner filter effect on the BTC absorption, achieving high response to 5-mC in a very short time (20 s). Moreover, such an efficient, instantaneous response probe presents excellent specificity towards 5-mC among various coexisting species in urine, and high sensitivity with a linear detection range of 0–200 μM and limit of detection (LOD) of 0.18 μM, thereby enabling it to be applied in accurately quantifying 5-mC content in real urine samples with high recoveries (95.5–98.5 %) and low relative standard deviations (< 2.5 %). This study demonstrates a simple, fast and effective approach for sensitive identification of urinary 5-mC, prefiguring its great potential for the assessment of DNA methylation associated with genetic diseases and tumors.
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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纳米技术
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生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
ELECTROANALYSIS
·
2023
·Vol. 35
·期 9
·e202300052
·
IF 3
Qiaoyu Ding,Xi Bai,Jiali Ju,Jiajun Shi,Guixin Li
A sensitive electrochemiluminescence (ECL) biosensor for the specific DNA sequence of hepatitis C virus (HCV) was developed based on the efficient quenching effect of the ferrocene cluster functionalized gold nanoparticles (Fc@AuNPs) on the ECL of electrodeposited silica@Ru(bpy) 3 2+ -chitosan-graphene oxide nanocomposite (SiO 2 @Ru−CS−GO). Graphene oxide (GO) can accelerate electron transfer rate, thus improving the ECL of Ru(bpy) 3 2+ on electrode surface. The molecular beacons (MB) was fixed to SiO 2 @Ru−CS−GO by glutaraldehyde (GA) using the Schiff reaction between amino groups of chitosan (CS) and MB. The ECL of SiO 2 @Ru−CS−GO was depressed greatly by the Fc@AuNPs labelled at the end of MB, then, a stronger ECL was observed when the distance between Fc@AuNPs and SiO 2 @Ru−CS−GO increased after the hybridization of target DNA with MB. Under optimum conditions, the restored ECL intensity increased linearly with the target DNA concentration in the range of 1.0×10 −16 ∼1.0×10 −10 mol ⋅ L −1 , and the limit of detection (LOD) is 1.4×10 −17 mol ⋅ L −1 . The proposed method exhibits acceptable stability and reproducibility. In general, the constructed HCV biosensor can be used for the sensitive detection of HCV in human serum, suggesting potential application prospects in bioanalysis.
研究领域:
分析化学
·
生物传感器
·
医学检测
✓ 已验证
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
·
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
·
分子诊断
·
食品监测
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
Analytical Methods
·
2019
·Vol. 12
·期 4
·491-498
·
IF 2.378
Zhang-Jin He,Tian-Fang Kang,Li-Ping Lu,Shui-Yuan Cheng
In this study, a novel electrochemiluminescence (ECL) biosensor based on a CdSe@CdS quantum dot (QD)-functionalized MoS2-modified electrode was developed for the sensitive detection of mercury ions. Polycationic poly(diallyldimethylammonium chloride) (PDDA) and negatively charged QDs were adsorbed on the surface of MoS2, in sequence, to form the QD–PDDA–MoS2 composites, which were employed as matrices for immobilizing thymine-rich DNA sequences (DNA1). The gold nanoparticles (AuNPs) were combined with another type of thymine-rich DNA sequences (DNA2) and glucose oxidase (GOD) to prepare the GOD–AuNP–DNA2 conjugates. The composites were characterized via UV-visible absorption spectroscopy, fluorescence and transmission electron microscopy. A thymine–Hg2+–thymine complex was formed through Hg2+ mismatching with DNA1 and DNA2. GOD was, therefore, modified onto the electrode surface. GOD catalyzed the reduction of dissolved oxygen by glucose to produce hydrogen peroxide, which was a co-reactant of the QDs. The electrochemiluminescence signal of the biosensor increased linearly with the increase in the Hg2+ concentration over the range from 1.0 × 10−12 to 1.0 × 10−6 M with a detection limit of 1 × 10−13 M. The reproducibility, stability and specificity of the biosensor was also studied. The biosensor was used to determine Hg2+ in a real water sample and satisfactory results were obtained.
研究领域:
分析化学
·
环境科学
·
生物传感技术
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 186
·期 1
·1-9
·
IF 5.705
Zhang Xingxing,Liu Qiang,Jin Yan,Li Baoxin
A fluorometric method is described for the determination of the activity of the enzyme T4 polynucleotide kinase phosphatase (T4 PNKP). A short 3′-terminus phosphorylated DNA strand is hybridized with a long DNA strand to produce a partially double-stranded DNA (dsDNA) substrate. On addition of T4 PNKP, the substrate is dephosphorylated to generate the long dsDNA, and then the long dsDNA acted as a template for synthesizing copper nanoclusters (CuNCs). The dsDNA-templated CuNCs display fluorescence with excitation/emission peak wavelengths of 340/570 nm. The fluorescence is DNA sequence-dependent. A DNA substrate was designed to enhance fluorescence and to reduce the background in order to improve the sensitivity of the assay. The assay has an analytical range that extends from 0.07 U mL −1 to 15 U mL −1 and a detection limit of 0.06 U mL −1 .
The sequence-dependent fluorescence of DNA-templated copper nanoclusters, which are in-situ synthesized through the reduction of CuSO 4 by ascorbate (Vc) in the presence of dsDNA template, is utilized to obtain the method for sensitive detection of T4 PNKP activity with near-zero background.
研究领域:
生物化学
·
分子生物学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Siqi Cai,Dejun Li,Fang Luo,Zhenyu Lin,Aiwen Huang,Bin Qiu
In this work, an electrochemiluminescence (ECL) biosensor for T-2 toxin detection has been designed by combining the high specificity of a DNA hydrogel system with the convenience and sensitivity of homogeneous ECL detection. Specifically, A hydrogel was engineered through the reaction between the carboxyl group of hyaluronic acid (HA) and the amino groups of both polyethyleneimine (PEI) and DNA-modified strands, resulting in a three-dimensional network that encapsulates glucose oxidase. Glucose oxidase (GOD) is capable of catalyzing the conversion of glucose into hydrogen peroxide (H₂O₂), which is a co-reactant necessary for the chemiluminescent reagent luminol to emit light. Upon the presence of the target analyte T-2 toxin, the aptamer recognizes and binds to it, leading to the dissociation and cleavage of the hydrogel framework. This process results in the leakage of the encapsulated GOD into the supernatant. Subsequently, Adding the collected supernatant to a phosphate-buffered saline (PBS) solution containing glucose and luminol for ECL testing yields a noticeable signal change. This sensor eliminates the need for electrode modification, simplifying the operation. Within a concentration range of 10 fg/mL to 100 ng/mL, the ECL signal intensity exhibits an excellent linear relationship with the logarithm of the target concentration, with a detection limit as low as 0.32 fg/mL. The method also demonstrates excellent reproducibility and stability. The sensor demonstrated precise detection capabilities in actual milk and beer samples, a crucial aspect for research focused on specific targets.
研究领域:
分析化学
·
生物技术
·
毒理学
·
食品安全
✓ 已验证
产品关联
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Dongxia Li,Yanan Peng,Liangqing Lu,Delun Chen,Bingrong Wang,Qiumei Pu,Qionglin Zhou,Xinxin Xiao,Mingpeng Fu,Xiaohong Wang,Jinchun Tu,Hua Pei,Qiang Wu
Adenosine triphosphate (ATP), as a crucial signaling molecule, is of high potential diagnosis value for multiple diseases. However, current technologies for ATP detection struggle to satisfy the harsh conditions required for biospecimen tests, existing several limitations for clinical practical application. In this study, the DNA tile response system was designed and constructed by integrating nucleic acid aptamer, hybrid chain reaction (HCR), and DNA tile self-assembly technology to achieve accurate detection of ATP in body fluid specimens. The DNA tile response system, possessing high-order structure characteristic, could remove HCR false-positive products and conquer the high noise background to enhance the detection sensitivity. Under the optimal reaction parameters, this system achieved the low limit of detection (LOD) of 189.23 nM over a wide linear range and splendid detection specificity faced with multiple interference terms. Furthermore, this system, possessing simple storage conditions, long-term detection stability, and superior anti-interference performance, was sufficient to satisfy the detection requirements for complicated biological specimens in multi-scenarios. Ultimately, the DNA tile response system showed excellent diagnostic efficiency that could easily distinguish urinary tract infection patients and healthy controls through ATP quantification, as well as hypertension patients and healthy ones. This system provides a high-potential rapid detection tool for early diagnosis and dynamic monitoring of urinary tract infections and hypertension, holding enormous potential for clinical translation due to the superiorities of simple operation, general applicability, and low instrument requirements.
研究领域:
生物技术
·
医学诊断
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）