SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Qingteng Lai,Yanke Zhang,Wei Chen,Zhengchun Liu
In this study, an ultrasensitive electrochemical-detection system based on a target “turn-on” signal-enhancement pseudocapacitor was established for microRNA-141 (miRNA-141) detection. A three-dimensional porous film was constructed on the electrode surface in situ by chitosan, catechol and nanomaterials (gold nanoparticles and nitrogen-doped carbon nanotubes). When miRNA-141 hybridized with the probe on the electrode, ferrocene (Fc) was specifically introduced into the electrode surface, and then a pseudocapacitor containing Fc, ruthenium ion (Ru 3+ ), catechol, and electrode formed (“turn on”). Fc and Ru 3+ consumed in the electrode reaction can quickly recover to the original valence state on the surface of catechol/quinone and again participated in the electrode reaction. A "one to many" relationship existed between the target and the signal molecule Fc, which contributed to the improvement of the sensor sensitivity. The signal strength and stability of Fc improved and thus improved the sensitivity and stability of miRNA-141 detection. The concentration information of miRNA-141 was collected from the change in Fc signal strength. The linear detection range was 0.5 fM to 800 pM, and the detection limit can be calculated as 43.73 aM. This system overcame the problems of low reliability and weak signal of the traditional electroactive-ion labeling method, which relied on the change in electrostatic attraction/repulsion before and after probe–target hybridization.
研究领域:
生物化学
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纳米技术
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电化学
·
生物传感器
·
癌症诊断
✓ 已验证
产品关联
DOI
高影响因子
Theranostics
·
2019
·
IF 8.063
Weipan Peng,Qian Zhao,Minghui Chen,Jiafang Piao,Weichen Gao,Xiaoqun Gong,Jin Chang
The accurate and quantitative detection of microRNAs (miRNAs) as next-generation, reliable biomarkers will provide vital information for cancer research and treatment. However, their unique, intrinsic features pose quite a challenge for miRNA profiling, especially for multiplexed detection. Thus, there is a strong and an ever-growing need to develop an accurate, simple, sensitive and specific miRNA sensing method. Methods: In this study, a simple and novel sensor is presented that uses a flow cytometry (FCM) method based on the double key “unlocked mechanism” and a fluorescence enrichment signal amplification strategy. The “unlocked mechanism” was cleverly designed via using hairpin DNA probes (HDs) labeled by fluorescent particles (FS) as the lock to block part of them, which can specifically hybridize with the probe on polystyrene microparticles (PS). The target miRNA and duplex-specific nuclease (DSN) forming the double key can specifically open the HDs and cleave a single-stranded DNA (ssDNA) into DNA/RNA dimers circularly in order to unlock the special part of the HDs to be specially enriched further on the PS. Results: The designed sensor with a hairpin structure and DSN special performance was found to have a high specificity. The circularly unlocking fluorescent probes and fluorescent signal enrichment can be beneficial for achieving a high sensitivity with a detection limit of 3.39 fM for miRNA-21. Meanwhile, the performance of multiplexing was estimated by simultaneous detection of miR-21 and miR-141, and the method also allowed for miR-21 detection in breast cancer blood samples. Conclusion: The designed sensor based on an “unlocked mechanism” and a signal enrichment strategy resulted in a one-pot, highly specific and sensitive detection of multiplex miRNAs. The whole detection without the need for a complex purification process is based on a FCM and is expected to have a great value in cancer diagnosis and biomedical research.
研究领域:
生物标志物检测
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癌症研究
·
生物医学工程
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
ELECTROCHIMICA ACTA
·
2022
·
IF 7.336
Qi Kang,Yuan Huang,Xuemei Ma,Mengmeng Li,Cheng Ma,Dazhong Shen
The electrochemiluminescence (ECL) spectral information is useful in ECL mechanism investigations, ratiometric ECL sensors and multi-analyte determinations. To overcome the shortcomings of low sensitivity in charge coupled devices (CCD) ECL spectrometer and low time efficiency for spectrum discrimination by using single band-pass filter (BPF) in one potential scan, a simple and sensitive ECL spectrum measurement platform is developed by placing a portable BPF array turntable device (with eleven measurement channels and one indicator channel) between the working electrode and photomultiplier in a conventional ECL analyzer. The systematic error due to the time dependent ECL intensity, which is the main obstacle for ECL spectrum measurement in wavelength scan mode, is neutralized by using the accumulated intensity for each channel in a group of spectrum measurement cycles. In a spectrum-resolved ratiometric ECL sensor for miroRNA-141 determination, near-infrared ECL emitter of methionine-stabilized Au nanoclusters (Met-Au NCs) is used as the tag and blue ECL emitter of g-C 3 N 4 as the reference. The ratio of the total ECL intensity in near-infrared wavelength region (from Met-Au NCs) to that in blue wavelength region (from g-C 3 N 4 ) is in linear correlation with the logarithmic concentration of microRNA-141 from 1 fM to 10 pM, with the limit of detection of 0.22 fM.
研究领域:
分析化学
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生物医学工程
·
分子生物学
✓ 已验证
DOI
高影响因子
MICROCHIMICA ACTA
·
2019
·Vol. 186
·期 8
·1-7
·
IF 5.479
Li Mei,Xu Xiong,Cai QingYou,Luo XuJian,Zhou ZhongGao,Xu GuoHai,Xie YongRong
A graphene oxide-based method has been developed for ultrasensitive and selective determination of microRNA-141 by means of rolling circle amplification (RCA) and exonuclease III (Exo III)-assisted recycling amplification. The method uses (a) a padlock probe with a hybrid sequence that is complementary to the target microRNA-141 at both the 5'- and the 3'-end, and (b) a long binding region of a signalling reporter strand. On addition of microRNA-141, it acts as the primer for triggering the RCA reaction following ligation. This results in the formation of a repeatedly concatenated sequence of the padlock probe. Subsequently, the RCA product hybridizes with the fluorescein-labelled signal strand to form the duplex DNA containing the blunt 3'-termini of signal strand. Addition of Exo III causes signal strand digestion and leads to RCA product recycling and liberation of fluorescein. Added graphene oxide does not absorb the fluorescein liberated because of the poor mutual interaction. Therefore, microRNA-141 can be quantified by measurement of the green fluorescence under excitation/emission wavelengths of 470/520 nm. The method has a 100 aM detection limit towards microRNA-141 and works in the wide range from 1 fM to 1 nM. It can discriminate even single-mismatched microRNA and shows good selectivity and sensitivity when applied to spiked human serum.
Schematic representation of a graphene oxide (GO)-based method for fluorometric determination of microRNA by using rolling circle amplification and exonuclease III (Exo III)-aided recycling amplification. With microRNA, the fluorescein-labelled signal strand becomes digested, and this genererates a fluorescent signal.
研究领域:
分子生物学
·
生物化学
·
纳米医学
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Lili Zhou,Ting Wang,Yan Bai,Yi Li,Juhui Qiu,Wen Yu,Sheng Zhang
s Rapid and efficient detection of tumor marker at the early stages is one of the crucial challenges in cancer diagnostics and therapy. In this study, an ultrasensitive electrochemical biosensor was fabricated by dual-amplified strategy for the detection of ultra-trace microRNA-141 (miRNA-141). Firstly, two split sequences contained G-quadruplex were connected by click chemistry-mediated nucleic acid strands self-assembly and the obtained complete G-quadruplex was complementary with miRNA-141 to formed DNA-RNA hybrid duplexes. Subsequently, the formed DNA-RNA hybrid duplexes were specifically recognized by duplex-specific nuclease (DSN), and the DNA part of the duplexes were cleaved and the miRNA-141 were released to trigger next cycle, which acquired a primal signal amplification by enzyme-assisted target recycling (EATR). Moreover, amino and thiol group multi-labeled functionalized fullerene nanoparticles (FC60) with a larger surface active sites and better biocompatibility, were designed rationally to modify the Au electrodes, which produced multiply-enhanced amplified signal. This dual-amplified sensing system exhibited a remarkable analytical performance for the detection of miRNA-141 in concentrations ranging from 0.1 pM to 100 nM and the detection limit of 7.78 fM was obtained. Compared with the biosensor with single amplification strategy such as EATR, this electrochemical biosensor based on dual-amplified strategy exhibited an excellent discrimination capability and higher analytical performance. Therefore, this electrochemical biosensor might hold a great potential for further applications in biomedical research and early clinical diagnosis.
研究领域:
生物医学工程
·
分子诊断学
·
癌症早期检测
✓ 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）