Scientific Reports
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2020
·Vol. 10
·期 1
·1-8
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IF 3.998
Tan Xiao-Hui,Li Yu-Bin,Liao Yan,Liu Hua-Zhong
In this work, an enzyme-free fluorescence resonance energy transfer (FRET) strategy was established for rapid and specific detection of the DNA sequence from Vibrio parahaemolyticus (VP) using hybridization chain reaction (HCR) amplification and triplex DNA. The triplex forming oligonucleotide (TFO) was labelled with carboxyfluorescein (FAM) as fluorescence donor, and hairpin sequence H1 was labelled by tetramethylrhodamine (TAMRA) as fluorescence receptor. In the present target VP DNA, the hairpin structure of molecular beacon (MB) was opened, the free end was released and hybridized with H1-TAMRA, and the HCR reaction was triggered by the alternate supplementation of H1-TAMRA and H2 to produce the notch double helix analogue. After the addition of TFO-FAM, a triplex structure was formed between HCR products (H1-TAMRA/H2) and TFO-FAM. A close contact between the donor and the receptor resulted in FRET. Under the optimal conditions, the fluorescence quenching value was inversely proportional to the concentration of target VP DNA in the range of 0.1–50 nmol L −1 , and the detection limit was 35 pmol L −1 .
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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微生物学
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PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2021
·
IF 6.558
Yawen Wu,Xiaolong Chen,Xiaogang Luo,Mei Yang,Changjun Hou,Danqun Huo
Carcinoembryonic antigen (CEA) is a key serum tumor marker which is overexpressed in all types of adenocarcinomas. Therefore, establish the ultrasensitive, accurate and rapid method for CEA detection is essential for reducing the mortality of cancer. Here, a bimetallic organic framework Cu/UiO-66 was synthesized through the simple two-step hydrothermal method and used to construct a “fluorescence turn-on” analytical method for CEA detection. Cu/UiO-66 can adsorb CEA aptamers modified with FAM (CEA/FAM-Apt) and take place photoinduced electron transfer (PET) between Cu/UiO-66 and FAM, resulting in the fluorescence of the FAM is quenched. When CEA is present, CEA and CEA/FAM-Apt are tightly combined, making CEA/FAM-Apt far away from the Cu/UiO-66 surface. As a result, the fluorescence intensity of the system was significantly restored. Under optimal conditions, the proposed “fluorescence turn-on” method can detect CEA as low as 0.01 ng mL −1 in a range of 0.01–0.3 ng mL −1 . Besides, this analytical method owns good selectivity , reproducibility and serum applicability, which provides a new platform for the direct detection of clinical diagnosis-related markers.
研究领域:
分析化学
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生物医学工程
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癌症诊断
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材料科学
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生物化学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2025
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IF 4.6
Anjie Guo,Yu Wu,Yunkai Xie,Wang Guo,Yiqing Guo,Xinai Zhang,Wen Zhang,Xiaobo Zou,Zongbao Sun
Di-(2-ethylhexyl) phthalate (DEHP) can leach into food and the environment, posing health and ecological risks. This paper introduces a novel CRISPR-based fluorescent aptasensor for on-site DEHP detection. The aptasensor selectively binds to DEHP, triggering a competitive displacement reaction that releases an Aptamer-dsDNA probe. After magnetic separation, the supernatant is analyzed via a Cas12a reporter system, where Cas12a activation cleaves a FAM-BHQ reporter, generating fluorescence. Combined with smartphone imaging, this method enables rapid result acquisition. The aptasensor shows high selectivity and sensitivity, detecting DEHP from 1 pg/mL to 1 μg/mL, with a lower limit of 0.15 pg/mL. It effectively detects DEHP in various real samples, offering reliable visual monitoring. This method offers a rapid and effective on-site detection strategy for food safety and environmental pollution monitoring.
研究领域:
食品安全
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环境监测
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生物传感器
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分子诊断
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CRISPR技术
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2018
·Vol. 185
·期 10
·1-6
·
IF 5.705
Ma Liang,Guo Ting,Pan Shuli,Zhang Yuhao
A fluorometric patulin (PAT) assay is presented that is based on the use of magnetic reduced graphene oxide (rGO) and DNase I. The fluorescence of the PAT aptamer labelled with 6-carboxyfluorescein (FAM) is quenched by magnetized reduced graphene oxide (rGO-Fe 3 O 4 ) due to fluorescence resonance energy transfer (FRET). However, in the presence of PAT, the labelled aptamer is stripped off from rGO-Fe 3 O 4 . The rGO-Fe 3 O 4 is then magnetically separated so that the fluorescence of free labelled PAT aptamer is restored. DNase I cannot hydrolyze the aptamer on rGO-Fe 3 O 4 , but it can cleave the free aptamer-PAT complex. This will release FAM and PAT which can undergo a number of additional cycles to trigger the cleavage of abundant aptamer. Recycling of DNase I-assisted target therefore leads to a strong amplification of fluorescence and consequently to an assay with low limit of detection. The detection limit for PAT is as low as 0.28 μg L −1 which is about 13 times lower than that without using DNase I. The method offers a new approach towards rapid, sensitive and selective detection based on an aptamer. Conceivably, it has a wide scope in that it may be applied to numerous other analytes if appropriate aptamers are available.
Schematic of a fluorometric assay based on the use of magnetic graphene oxide and DNase I. It was applied to the determination of patulin. DNase I was introduced for recycling amplification. The detection limit is about 13 times lower than that without using DNase I. Figure a contains poor quality of text in image. Otherwise, please provide replacement figure file.Thank you. I will provide the figure file.
研究领域:
分析化学
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食品安全
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环境科学
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生物传感器
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DOI
PubMed
高影响因子
ACS Applied Nano Materials
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2021
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IF 5.097
Li Ping Cao,Yao Wang,Yan Bai,Yong Jian Jiang,Chun Mei Li,Hua Zuo,Yuan Fang Li,Shu Jun Zhen,Cheng Zhi Huang
Inspired by the specific optical properties of DNA photonic nanowires (PNs) on which fluorescent dyes are to be precisely positioned and photonic energy transfer over nanometers can be achieved, herein, we develop a DNA PN-based homogeneous entropy-driven biomolecular assay (HEBA). Using homogeneous Förster resonance energy transfer (FRET) of Cy3, we constructed a DNA PN capable of transferring energy from the green donor dye (FAM) to the red acceptor fluorophore (Cy5) across a total distance of 10.2 nm with an end-to-end transfer efficiency of 13.2%. By taking DNA PNs as the substrate of entropy-driven circuit, thrombin was detected as a proof-of-principle target. The trigger of HEBA by thrombin, during which separation of Cy3, FAM, and Cy5 occurred and the FRET of the PN got interrupted, resulted in a high value of signal-to-background ratio (S/B, signal and background were FFAM/FCy5 in the presence and absence of thrombin, respectively), giving a detection dynamic range of 5–25 nM with a detection limit of 2.7 nM. This strategy offers an alternative platform for biomarker clinical diagnosis, greatly expanding the application fields of DNA PNs to analytical chemistry.
研究领域:
生物技术
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分析化学
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纳米技术
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分子生物学
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产品关联
DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Ming Li,Weipeng Teng,Wenying Lu,Mingna Sun,Jinsheng Duan,Xuchun Qiu
The improvement of dual-mode techniques was of particular interest to researchers, which might enhance the detection performance and applicability. Here, a dual-mode optical aptasensor (DO-aptasensor) platform based on exonuclease I (Exo I) cyclic digestion and synergistic enhancement strategy had proposed for zearalenone (ZEN). Following the preparation of dumbbell-shaped signal probe, the Exo I-based cyclic digestion amplification performed, and then the synergistic enhancement effect carried out to achieve the Poly-HRP-based colorimetry and FAM-SGI-based fluorescence. The efficient homogeneous system realized through the magnetic separation, while the signal interference further eliminated by the graphene oxide (GO). The LOD values were as low as 0.067 ng mL −1 for colorimetry mode and 0.009 ng mL −1 for fluorescence mode, which reduced 23-fold and 172-fold than ELASA by same ZEN-Apt. This promising platform gave rise to a dual-mode optical readout, improved sensitivity and positively correlated detection. Meanwhile, the DO-aptasensor also exhibited the acceptable specificity, desirable reliability and excellent practicability. This novel avenue of aptasensor platform hold great potential for dual-mode optical monitoring of other targets, which can further expand the application scope of Exo I-based signal amplification and synergistic enhancement effect.
研究领域:
分析化学
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生物传感器
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食品安全
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分子生物学
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DOI
PubMed
高影响因子
Analytical Methods
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2024
·
IF 2.7
Xiaoxia Lu,Wenlong Lu,Dong Hua
In the study, a novel surface-enhanced Raman scattering (SERS)-lateral flow assay (LFA) tray for the real-time detection of pyroptosis-associated miR-155-5p in breast cancer cells was established and validated. The SERS probe modified with monoclonal antibodies and functionalized HP1@5-FAM was first synthesized. When miR-155-5p was present, HP1@5-FAM on the SERS probe specifically recognized target miRNAs and hybridized with them, resulting in HP2 on the T line only capturing some SERS probes that were not bound to miR-155-5p. The T line appeared as a light orange band or there was no color change, and the corresponding Raman detection result showed a weak or insignificant Raman signal. The SERS probe showed high selectivity, satisfactory stability, and excellent reproducibility, and the limit of detection (LOD) for miR-155-5p was 7.26 aM. Finally, the proposed SERS-LFA tray was applied to detect miR-155-5p in MBA-MD-468 cells that underwent varying degrees of pyroptosis, and the detection results of SERS were consistent with those of the conventional real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. The study demonstrated that the SERS-LFA tray was a convenient and ultrasensitive method for miR-155-5p real-time detection, which could provide more detailed information for pyroptosis and be of potential value in guiding the treatment of breast cancer.
研究领域:
癌症生物学
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分子生物学
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生物医学工程
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分析化学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Yuanyi Wu,Yan Yang,Shihao Yang,Yang Ju,Yang Jiang,Yumei Xiao,Jialin He,Tian Liu,Leyao Tang,Kaiyong Wang,Yi Yang
Interfacial and interlayer interactions between layered double hydroxides (LDHs) and nucleic acids offer LDHs great potential to absorb and detect nucleic acids. Herein, we demonstrate a simple, rapid, and versatile fluorescent sensing platform based on two-dimensional LDHs and exonuclease I (Exo I) for the selective analysis of DNA and microRNA. ZnNiAl-LDHs were utilized as quenchers for FAM labelled on single-stranded DNA (ssDNA) probes via dynamic quenching with high fluorescence quenching ability. The quenching mechanism was explored by means of spectral analysis, characterization experiments, DNA adsorption and desorption studies, zeta potential analysis, and molecular dynamics simulations. The driving forces for the adsorption of LDHs toward fluorophore-labelled nucleic acids were confirmed to be electrostatic interaction, van der Waals force, and anion exchange for the first time. In order to distinguish the formed target/probe duplexes from FAM-ssDNA probes, Exo I was introduced to specifically hydrolyze FAM-ssDNA probes. The fabricated fluorescent sensing platform for nucleic acids detection shows a low detection limit of 0.08 nM. Finally, this sensor was effectively utilized for the detection of DNA and microRNA in actual biological and environmental samples, achieving recoveries ranging from 96.4 % to 105 %, indicating high accuracy. This work proposed a promising sensing platform for sequence-specific nucleic acids detection, and provides a new perspective on two-dimensional LDHs as structural supports and biomolecular reservoirs for biomedical applications.
研究领域:
生物技术
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纳米技术
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分析化学
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分子生物学
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2022
·
IF 9.221
Liping Zhao,Irfan Muhammad,Xiaomin Zhang,Ge Yang,Yao Ma,Bo Wei,Feng Qu
A new apta-sensing probe based on magnetic hydrophilic polymer (MHP) was proposed for simple and sensitive detection of fetuin-A in serum matrix. Briefly, MHP was the hydrophilic polymer modified magnetic nanoparticles , which was synthesized through free radical polymerization method with hydrophilic monomers and magnetic nanoparticles. Then, aptamer against fetuin-A first selected through capillary electrophoresis-SELEX was hybridized with FAM-labeled aptamer complementary sequence (CS) to form duplex structure, which was finally conjugated on MHP to form MHP apta-sensing probe (MHPA). In this probe, MHP with hydrophilic property was employed as separation carrier, aptamer was used for fetuin-A binding and FAM-labeled CS as signal probe . With the presence of fetuin-A, CS escaped and could be detected because of the competitive binding between fetuin-A-aptamer and aptamer-CS. Because of the synergistic effect of specific affinity of aptamer on protein and hydrophilic property of the polymer, MHPA exhibited sensitive and selective detection for fetuin-A. Under optimal conditions, the linear range was 5.0 nM-4.0 μM with the detection limit of 4.4 nM. What’s more, MHPA engaged with good stability and selectivity had been successfully applied in the detection of fetuin-A in serum samples. The as prepared hydrophilic polymer assistant apta-sensing probe with facile and sensitive properties, provides novel explication for efficient protein detection.
研究领域:
分析化学
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生物传感器
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蛋白质检测
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纳米技术
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DOI
高影响因子
TALANTA
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2024
·
IF 5.6
Chenxu Peng,Mingyu Leng,Yongguang Gao,Qiumei Feng,Xiangmin Miao
In situ monitoring of intracellular microRNAs (miRNAs) often encounters the challenges of surrounding complexity, coexistence of precursor miRNAs (pre-miRNAs) and the degradation of biological enzyme in living cells. Here, we designed a novel probe encapsulated DNA tetrahedral molecular sieve (DTMS) to realize the size-selective detection of intracellular miRNA 21 that can avoid the interference of pre-miRNAs. In such strategy, quencher (BHQ-1) labeled probe DNA (S6-BHQ 1) was introduced into the inner cavity of fluorophore (FAM) labeled DNA tetrahedral scaffolds (DTS) to prepare DTMS, making the FAM and BHQ-1 closely proximate, and resulting the sensor in a "signal-off" state. In the presence of miRNA 21, strand displacement reaction happened to form more stable DNA double-stranded structure, accompanied by the release of S6-BHQ 1 from the inner cavity of DTMS, making the sensor in a "signal-on" state. The DTMS based sensing platform can then realized the size-selective detection of miRNA 21 with a detection limit of 3.6 pM. Relying on the mechanical rigidity of DTS and the encapsulation of DNA probe using DTMS, such proposed method achieved preferable reproducibility and storage stability. Moreover, this sensing system exhibited good performance for monitoring the change of intracellular miRNA 21 level during the treatment with miRNA-related drugs, demonstrating great potential for biological studies and accurate disease diagnosis.
研究领域:
分子生物学
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生物技术
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纳米技术
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细胞生物学
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分析化学
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医学诊断
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DOI
PubMed
高影响因子
TALANTA
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2024
·
IF 5.6
Yingao Yang,Ruipeng Chen,Yifen Guo,Jiaxin Zhang,Shuyue Ren,Huanying Zhou,Zhixian Gao
A two-color fluorescent sensing strategy based on a functionalized tetrahedral DNAzyme nanotweezer (FTDN) was developed to detect ochratoxin A (OTA) utilizing the multifunctional properties of DNA nanotechnology. The FTDN enables rapid OTA detection directly through a Cy5 fluorescent group, modified to respond to the target signal. Additionally, FTDN exhibits DNAzyme cutting activity in the presence of Mg 2 ⁺ ions, enabling it to traverse DNA nanoflower-functionalized magnetic beads. This process results in the continuous cleavage of DNA nanoflowers labeled with numerous FAM fluorescent groups, thereby amplifying the detection signal and enhancing OTA sensitivity. The linear ranges for the Cy5 and FAM signals in response to OTA were 5–1000 ng/mL and 0.05–100 ng/mL, respectively, with corresponding limits of detection (LOD) of 1.59 ng/mL and 0.03 ng/mL. This study demonstrates that dual-color fluorescence via Cy5 and FAM can effectively verify OTA detection in food, significantly reducing false-positive and false-negative rates. The proposed platform offers sensitive and accurate detection of mycotoxins in food and can be adapted for monitoring other trace contaminants by simply altering the aptamer.
研究领域:
分析化学
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纳米技术
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食品科学
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毒理学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
·
IF 12.545
Xiaojuan Wang,Zefeng Mao,Ruipeng Chen,Shuning Li,Shuyue Ren,Jun Liang,Zhixian Gao
DNA origami technology has great potential for biosensor applications. Here, we described the construction of a self-assembled DNA origami biosensor for the precise localization of fluorescent aptamers . Due to the molecular weight difference between DNA origami and aptamer, centrifugal filters were used to quantitatively detect adenosine triphosphate (ATP). The ATP-specific aptamer labeled with fluorescence reporter 6-carboxyfluorescein FAM (FAM-aptamer) was selected as the recognition element and signal probe . ATP duplexed aptamers bound to triangular DNA origami by base-complementary pairing, resulting in high fluorescence signals on the origami arrays. The competitive binding of ATP toward the FAM-aptamer triggered the release of FAM-aptamer-ATP complexes from the surface of the origami array, resulting in weakened fluorescence signals. For ATP quantification, 100 kD centrifugal filters were employed, followed by measurement of the fluorescence signal trapped on the origami arrays of the filter device. The successful synthesis of origami-aptamer arrays was characterized by atomic force microscopy , laser confocal microscopy , and electrophoresis . Fluorescence measurements exhibited an excellent linear relationship with logarithms of ATP concentrations within 0.1–100 ng mL −1 , with a detection limit of 0.29 ng mL −1 . By replacing aptamers and complementary strands, we demonstrated the potential of this method for 17β-estradiol detection. Considering that the detection mechanism is based on the hybridization and displacement of DNA strands, the detection system had the potential for recharging. Our study provides new insights into applying DNA origami technology in small molecule detection.
研究领域:
生物传感器
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纳米技术
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
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