The construction of a novel nucleic acids detection microplatform based on the NSET for one-step detecting TK1-DNA and microRNA-21

Zhang, Jian; Zhao, Qian; Wu, Yudong; Zhang, Bo; Peng, Weipan; Piao, Jiafang; Zhou, Yurui; Gao, Weichen; Gong, Xiaoqun; Chang, Jin

doi:10.1016/j.bios.2017.05.039

Cited by 14 publications

(6 citation statements)

References 40 publications

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“…To evaluate the practicability of the proposed method in biological samples, the target miR-21 was detected using a standard addition to spiked human serum samples, as performed in previous reports 36, 37. A final concentration (50 × 10 -12 M) of standard miRNA was added into five healthy serum samples for evaluation by our proposed method.…”

Section: Resultsmentioning

confidence: 99%

An innovative “unlocked mechanism” by a double key avenue for one-pot detection of microRNA-21 and microRNA-141

Peng¹,

Zhao²,

Chen³

et al. 2019

Theranostics

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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.

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Section: Resultsmentioning

confidence: 99%

An innovative “unlocked mechanism” by a double key avenue for one-pot detection of microRNA-21 and microRNA-141

Peng¹,

Zhao²,

Chen³

et al. 2019

Theranostics

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“…To test the applicability of the developed sensor for miRNA analysis in real samples, target miRNA-21 was monitored using standard addition in spiked serum as previous reports. − First, miRNA-21 with a concentration of (10 × 10 –10 M) was added into the serum samples obtained from five healthy volunteers, and then detected by our DSN-amplified PS@Au probe-based sensor. A determined content was obtained according to the calibration curve of miRNA-21 in Figure S5.…”

Section: Results and Discussionmentioning

confidence: 99%

Simple and Sensitive Quantification of MicroRNAs via PS@Au Microspheres-Based DNA Probes and DSN-Assisted Signal Amplification Platform

Zhao

Piao

Peng

et al. 2018

ACS Appl. Mater. Interfaces

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Identifying the microRNA (miRNA) expression level can provide critical information for early diagnosis of cancers or monitoring the cancer therapeutic efficacy. This paper focused on a kind of gold-nanoparticle-coated polystyrene microbeads (PS@Au microspheres)-based DNA probe as miRNA capture and duplex-specific nuclease (DSN) signal amplification platform based on an RGB value readout for detection of miRNAs. In virtue of the outstanding selectivity and simple experimental operation, 5'-fluorochrome-labeled molecular beacons (MBs) were immobilized on PS@Au microspheres via their 3'-thiol, in the wake of the fluorescence quenching by nanoparticle surface energy transfer (NSET). Target miRNAs were captured by the PS@Au microspheres-based DNA probe through DNA/RNA hybridization. DSN enzyme subsequently selectively cleaved the DNA to recycle the target miRNA and release of fluorophores, thereby triggering the signal amplification with more free fluorophores. The RGB value measurement enabled a detection limit of 50 fM, almost 4 orders of magnitude lower than PS@Au microspheres-based DNA probe detection without DSN. Meanwhile, by different encoding of dyes, miRNA-21 and miRNA-10b were simultaneously detected in the same sample. Considering the ability for quantitation, high sensitivity, and convenient merits, the PS@Au microspheres-based DNA probe and DSN signal amplification platform supplied valuable information for early diagnosis of cancers.

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“…These above-mentioned clinical demands require the NATs to be satisfied with high specificity, high sensitivity, as well as high simplicity, which, as a consequence, have driven the development of various nucleic acid detection techniques. For instance, researchers have developed signal amplification − and recognition interface engineering strategies for improving sensitivity, − used controllable hybridization approaches for improving specificity, − and used one-step design for improving simplicity (Figure b). − From the detection limit point of view, we found that the one-step-based analysis exhibited the highest detection limit in spite of its simplicity. The supplementary means including DNA nanosturcture-based regulation, nucleic acid hybridization, and amplification approaches are facilitated to improve the detection sensitivity.…”

Section: Clinical Translation-driven Natsmentioning

confidence: 93%

Nucleic Acid Tests for Clinical Translation

et al. 2021

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Nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are natural biopolymers composed of nucleotides that store, transmit, and express genetic information. Overexpressed or underexpressed as well as mutated nucleic acids have been implicated in many diseases. Therefore, nucleic acid tests (NATs) are extremely important. Inspired by intracellular DNA replication and RNA transcription, in vitro NATs have been extensively developed to improve the detection specificity, sensitivity, and simplicity. The principles of NATs can be in general classified into three categories: nucleic acid hybridization, thermal-cycle or isothermal amplification, and signal amplification. Driven by pressing needs in clinical diagnosis and prevention of infectious diseases, NATs have evolved to be a rapidly advancing field. During the past ten years, an explosive increase of research interest in both basic research and clinical translation has been witnessed. In this review, we aim to provide comprehensive coverage of the progress to analyze nucleic acids, use nucleic acids as recognition probes, construct detection devices based on nucleic acids, and utilize nucleic acids in clinical diagnosis and other important fields. We also discuss the new frontiers in the field and the challenges to be addressed.

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The construction of a novel nucleic acids detection microplatform based on the NSET for one-step detecting TK1-DNA and microRNA-21

Cited by 14 publications

References 40 publications

An innovative “unlocked mechanism” by a double key avenue for one-pot detection of microRNA-21 and microRNA-141

An innovative “unlocked mechanism” by a double key avenue for one-pot detection of microRNA-21 and microRNA-141

Simple and Sensitive Quantification of MicroRNAs via PS@Au Microspheres-Based DNA Probes and DSN-Assisted Signal Amplification Platform

Nucleic Acid Tests for Clinical Translation

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