Dual-probe fluorescent biosensor based on T7 exonuclease-assisted target recycling amplification for simultaneous sensitive detection of microRNA-21 and microRNA-155

Zheng, Yufeng; Chen, Jinyuan; Li, You; Xu, Yang; Chen, Li; Chen, Wei; Liu, Ailin; Lin, Xinhua; Weng, Shaohuang

doi:10.1007/s00216-020-03121-6

Cited by 25 publications

(13 citation statements)

References 44 publications

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“…As the antibodies show fluorescence after binding to fluorescence-labeled switching peptides, a quencher molecule was bound to the detection antibodies to absorb the fluorescence from the switching peptides before the binding of the antigens (target analytes). In this study, the switching peptide H2 (labeled with FAM) was complexed with antibodies against Inf-A and Inf-B, and graphene was labeled as a quencher to absorb the fluorescence from the switching peptide H2 [ 20 , 21 ]. As graphene could absorb the fluorescence signal in the visible range of 400–700 nm, the fluorescence from the labeled fluorescence dye, FAM (excitation and emission wavelengths of 488 and 514 nm, respectively), could be effectively quenched.…”

Section: Resultsmentioning

confidence: 99%

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

Kim

Bong

Shin³

et al. 2022

BioChip J

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One-step homogeneous immunoassay was developed for detecting influenza viruses A and B (Inf-A and Inf-B) using the switching peptide H2. As the fluorescence-labeled switching peptide dissociated from the binding pocket of detection antibodies, the fluorescence signal could be directly generated by the binding of Inf-A and Inf-B without washing (i.e., one-step immunoassay). For the one-step homogeneous immunoassay with detection antibodies in solution, graphene was labeled with the antibodies as a fluorescence quencher. To test the feasibility of the homogeneous one-step immunoassay, the stability of the antibody complex with the switching peptide was evaluated under different pH and salt conditions. The one-step homogeneous immunoassay with switching peptide was conducted using influenza virus antigens in phosphate-buffered saline and real samples with inactivated Inf-A and Inf-B spiked in serum. Finally, the one-step homogeneous immunoassay results were compared with those of commercially available lateral flow immunoassays.

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Section: Resultsmentioning

confidence: 99%

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

Kim

Bong

Shin³

et al. 2022

BioChip J

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“…High expression of miR-155 and miR-22 predicts recurrence in NSCLC [115,116]. Recently, a homogeneous T7 exonuclease (T7 Exo)-assisted signal amplification combined with GO quenching platform has been applied to detect miRNA-21 and miRNA-155 simultaneously with FAM and ROX-labelled single-strand DNA probes involved in drug resistance [117] LFIA immunoassays are a versatile tool in the field of point-of-care (POC) diagnostics. Routine LFIA test performs through antibodies.…”

Section: Fluorescence-based Biosensingmentioning

confidence: 99%

Liquid Biopsy-Based Biosensors for MRD Detection and Treatment Monitoring in Non-Small Cell Lung Cancer (NSCLC)

et al. 2021

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Globally, non-small cell lung cancer (NSCLC) is the leading cause of cancer deaths. Despite advancements in chemotherapy and targeted therapies, the 5-year survival rate has remained at 16% for the past forty years. Minimal residual disease (MRD) is described as the existence of either isolated tumour cells or circulating tumour cells in biological liquid of patients after removal of the primary tumour without any clinical signs of cancer. Recently, liquid biopsy has been promising as a non-invasive method of disease monitoring and treatment guidelines as an MRD marker. Liquid biopsy could be used to detect and assess earlier stages of NSCLC, post-treatment MRD, resistance to targeted therapies, immune checkpoint inhibitors (ICIs) and tumour mutational burden. MRD surveillance has been proposed as a potential marker for lung cancer relapse. Principally, biosensors provide the quantitative analysis of various materials by converting biological functions into quantifiable signals. Biosensors are usually operated to detect antibodies, enzymes, DNA, RNA, extracellular vesicles (EVs) and whole cells. Here, we present a category of biosensors based on the signal transduction method for identifying biosensor-based biomarkers in liquid biopsy specimens to monitor lung cancer treatment.

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“…In order to improve the analytical sensitivity in the quantification of miRNAs, signal amplification technologies have been introduced, such as rolling circle amplification (RCA), , exonuclease assisted amplification, , catalytic hairpin self-assembly (CHA), − hybrid chain reaction (HCR), , strand displacement reaction (SDR), , and deoxyribozyme (DNAzyme). , Multicomponent nucleic acid enzymes (MNAzyme) are derived from DNAzyme with higher selectivity and more flexible design . MNAzyme distributes the catalytic core into two partial enzymes (partzyme A and partzyme B), which are composed of a partial catalytic core, a substrate arm and a sensor arm.…”

Section: Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

DNA Tetrahedron-Based MNAzyme for Sensitive Detection of microRNA with Elemental Tagging

Liu

et al. 2021

ACS Appl. Mater. Interfaces

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Heterogeneous immunoassay based on magnetic separation is commonly used in inductively coupled plasma-mass spectrometry (ICP-MS)-based biomedical analysis with elemental labeling. However, the functionalized magnetic beads (MBs) often suffer from non-specific adsorption and random distribution of the functional probes. To overcome these problems, DNA tetrahedron (DT)-functionalized MBs were designed and further conjugated with substrate modified Au NPs (Sub-AuNP). Based on the prepared MB-DT-AuNP probes, an MB-DT based multicomponent nucleic acid enzyme (MNAzyme) system involving Au NPs as the elemental tags was proposed for highly sensitive quantification of miRNA-155 by ICP-MS. Target miRNA would trigger the assembly of MNAzyme, and Sub-AuNP would be cleaved from the MB-DT-AuNP probe, resulting in a cyclic amplification. Single-stranded DNA-functionalized MB (MB-ssDNA)-AuNP probes were prepared as well. Comparatively, the amount of Au NPs grafted onto MB-ssDNA-AuNP probes was higher than that grafted onto MB-DT-AuNP probes. Meanwhile, a higher signal-to-noise ratio was obtained by using MB-DT-AuNP probes over MB-ssDNA-AuNP probes in the MNAzyme system. Under the optimal experimental conditions, the limit of detection for target miRNA obtained by using MB-DT-AuNP probes was 1.15 pmol L–1, improved by 23 times over that obtained by the use of MB-ssDNA-AuNP probes. The proposed MB-DT-MNAzyme-ICP-MS method was applied to the analysis of miRNA-155 in serum samples, and recoveries of 86.7–94.6% were obtained. This method is featured with high sensitivity, good specificity, and simple operation, showing a great application potential in biomedical analysis.

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Dual-probe fluorescent biosensor based on T7 exonuclease-assisted target recycling amplification for simultaneous sensitive detection of microRNA-21 and microRNA-155

Cited by 25 publications

References 44 publications

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

One-Step Homogeneous Immunoassay for the Detection of Influenza Virus Using Switching Peptide and Graphene Quencher

Liquid Biopsy-Based Biosensors for MRD Detection and Treatment Monitoring in Non-Small Cell Lung Cancer (NSCLC)

DNA Tetrahedron-Based MNAzyme for Sensitive Detection of microRNA with Elemental Tagging

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