PCR-free and chemistry-based technology for miR-21 rapid detection directly from tumour cells

Delgado-González, Antonio; Robles-Remacho, Agustín; Marín-Romero, Antonio; Detassis, Simone; López‐Longarela, Bárbara; Lopez‐Delgado, Francisco Javier; Miguel-Perez, Diego de; Guardia-Monteagudo, Juan J.; Fara, Mario Antonio; Tabraue-Chávez, Mavys; Pernagallo, Salvatore; Sánchez‐Martín, Rosario M.; Díaz‐Mochón, Juan José

doi:10.1016/j.talanta.2019.03.039

Cited by 20 publications

(13 citation statements)

References 19 publications

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“…Copyright 2016 American Chemical Society. (B) Bio-labeling of a PNA:miR-21 duplex with a biotinylated reactive base described in [115]. (C) Detection of miR-208a using the SPR-based methodology, adapted from [44].…”

Section: Discussionmentioning

confidence: 99%

“…Delgado-Gonzalez et al recently proposed a detection system for single base resolution quantification of miR-21 levels in cell lysate, based on dynamic chemistry. By functionalization of magnetic nanospheres with a PNA containing an abasic site, they were able to induce a selective base coupling with a biotinylated reactive nucleobase (via reductive amination), templated by the target sequence [115]. In this particular case, as depicted in Figure 10B, this reductive amination is only possible in presence of the PNA backbone and cannot be applied to natural oligonucleotide probes.…”

Section: Other Methodologiesmentioning

confidence: 99%

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PNA-Based MicroRNA Detection Methodologies

2020

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MicroRNAs (miRNAs or miRs) are small noncoding RNAs involved in the fine regulation of post-transcriptional processes in the cell. The physiological levels of these short (20–22-mer) oligonucleotides are important for the homeostasis of the organism, and therefore dysregulation can lead to the onset of cancer and other pathologies. Their importance as biomarkers is constantly growing and, in this context, detection methods based on the hybridization to peptide nucleic acids (PNAs) are gaining their place in the spotlight. After a brief overview of their biogenesis, this review will discuss the significance of targeting miR, providing a wide range of PNA-based approaches to detect them at biologically significant concentrations, based on electrochemical, fluorescence and colorimetric assays.

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Section: Discussionmentioning

confidence: 99%

Section: Other Methodologiesmentioning

confidence: 99%

PNA-Based MicroRNA Detection Methodologies

2020

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“…The detection may be done with any methodology (chemiluminescent, fluorescent, or colorimetric), depending on the type of modification the SMART-NB carries. The technology may be used for direct detection from several sources, including biofluids, avoiding RNA extraction, RT and/or PCR amplification and it has been validated in several contexts for miRNAs analysis [155][156][157][158][159].…”

Section: Chem-natmentioning

confidence: 99%

Measurements Methods for the Development of MicroRNA-Based Tests for Cancer Diagnosis

Precazzini

Detassis²,

Imperatori

et al. 2021

IJMS

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Studies investigating microRNAs as potential biomarkers for cancer, immune-related diseases, or cardiac pathogenic diseases, among others, have exponentially increased in the last years. In particular, altered expression of specific miRNAs correlates with the occurrence of several diseases, making these molecules potential molecular tools for non-invasive diagnosis, prognosis, and response to therapy. Nonetheless, microRNAs are not in clinical use yet, due to inconsistencies in the literature regarding the specific miRNAs identified as biomarkers for a specific disease, which in turn can be attributed to several reasons, including lack of assay standardization and reproducibility. Technological limitations in circulating microRNAs measurement have been, to date, the biggest challenge for using these molecules in clinical settings. In this review we will discuss pre-analytical, analytical, and post-analytical challenges to address the potential technical biases and patient-related parameters that can have an influence and should be improved to translate miRNA biomarkers to the clinical stage. Moreover, we will describe the currently available methods for circulating miRNA expression profiling and measurement, underlining their advantages and potential pitfalls.

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“…This first step, to hybridise the miR-122, was performed in a 96-well plate using a microplate orbital at 700 rpm for 1 h at 40 • C. After the hybridization, the DGL-122 beads were washed three times with the wash buffer. The DGL-122 beads were resuspended in 50 µL of assay buffer containing 5 µM SMART-C biotin and 1 mM sodium cyanoborohydride [17][18][19][20][27][28][29][30][31][32][33]. The 96-well plate was shaken at 700 rpm at 40 • C for 1 h. The beads were washed three more times and incubated with 70 µL of 2 µg/mL SA-PE for 5 min at 40 • C while being shaken at 700 rpm.…”

Section: Calibration Curve For Mir-122 Assaymentioning

confidence: 99%

“…With the advances made by our team with dynamic chemical labelling (DCL) technology for the direct detection of nucleic acids, the deliverable of simultaneous detection of protein and miRNA in clinical samples is now possible. The DCL approach [17][18][19][20][27][28][29][30][31][32][33] is particularly well suited to deliver consistent and reliable quantitative readings of miRNAs in clinical samples when merged with bead-based systems. By simplifying the workflow, especially removing extraction, isolation and amplification steps, DCL was able to direct detect miRNAs in enzyme-linked immunosorbent assay (ELISA)-type format without affecting protein co-analytes, overcoming the current limitation issues that inhibit the development of simultaneous detection of proteins and miRNAs with high specificity and accuracy.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Detection of Drug-Induced Liver Injury Protein and microRNA Biomarkers Using Dynamic Chemical Labelling on a Luminex MAGPIX System

et al. 2021

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Drug-induced liver injury (DILI) is a potentially fatal adverse event and a leading cause for pre- and post-marketing drug withdrawal. Several multinational DILI initiatives have now recommended a panel of protein and microRNA (miRNA) biomarkers that can detect early liver injury and inform about mechanistic basis. This manuscript describes the development of seqCOMBO, a unique combo-multiplexed assay which combines the dynamic chemical labelling approach and an antibody-dependant method on the Luminex MAGPIX system. SeqCOMBO enables a versatile multiplexing platform to perform qualitative and quantitative analysis of proteins and miRNAs in patient serum samples simultaneously. To the best of our knowledge, this is the first method to profile protein and miRNA biomarkers to diagnose DILI in a single-step assay.

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PCR-free and chemistry-based technology for miR-21 rapid detection directly from tumour cells

Cited by 20 publications

References 19 publications

PNA-Based MicroRNA Detection Methodologies

PNA-Based MicroRNA Detection Methodologies

Measurements Methods for the Development of MicroRNA-Based Tests for Cancer Diagnosis

Simultaneous Detection of Drug-Induced Liver Injury Protein and microRNA Biomarkers Using Dynamic Chemical Labelling on a Luminex MAGPIX System

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