Ratiometric enhanced fluorometric determination and imaging of intracellular microRNA-155 by using carbon dots, gold nanoparticles and rhodamine B for signal amplification
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An ultrasensitive and highly reliable ratiometric assay is described for the determination of microRNA-155. It works at the attomolar concentration level and has high selectivity which warrants its potential application in cancer biomarker tracking. The excellent performance of this method results from (a) the use of a hybrid conjugate prepared from Rhodamine B (RhB), carbon dots (CDs) and probe-microRNA, and (b) from the measurement of fluorescence resonance energy transfer (FRET) that is observed in the AuNP/target-microRNA system as a result of RNA hybridization. The dye RhB (emission peak at 580 nm) serves as an internal reference. The sensitivity of this assay is increased by about 30% because of the broad emissions of CDs (489 nm and 665 nm) through a sequential FRET phenomenon. RhB-CDs were covalently bio-conjugated to probe microRNA. In the presence of AuNPs, the fluorescence of the CDs is quenched, while in the presence of microRNA-155, the ratio of fluorescences at 489 and 665 nm (I489/I665) is enhanced again. A linear relationship exists between the ratio of fluorescence and the concentration of microRNA-155 in the range from 1 aM to 0.1 μM, and the detection limit is 0.3 aM. The assay was applied to quantitative studies of target microRNA-155 in multiple pathways associated with cancer progression in biological fluids include human serum samples and cancer cells. The nanoprobe also deliver clear signal to microRNA target in fixed and lived MDA-MB-231 cells.
KeywordsHybridization Live cell Ratiometric sensing Dual emission Fluorescence resonance energy transfer
This research was supported by the Iranian Nanotechnology Initiative (142565) and the Research Offices of the University of Kurdistan (grant number 4.1404086).
Compliance with ethical standards
The author(s) declare that they have no competing interests.
- 7.Dong H, Zhang J, Ju H, Lu H, Wang S, Jin S, Hao K, Du H, Zhang X (2012) Highly sensitive multiple microRNA detection based on fluorescence quenching of graphene oxide and isothermal Strand- displacement polymerase reaction. Anal Chem 84:4587–4593. https://doi.org/10.1021/ac300721u CrossRefPubMedGoogle Scholar
- 10.Azimzadeh M, Rahaie M, Nasirizadeh N, Ashtari K, Naderi-Manesh H (2015) An electrochemical nanobiosensor for plasma miRNA-155, based on graphene oxide and gold nanorod, for early detection of breast cancer. Biosens Bioelectron 77:99–106. https://doi.org/10.1016/j.bios.2015.09.020 CrossRefPubMedGoogle Scholar
- 12.Mansouri-Majd S, Salimi A, Ghasemi F (2018) An ultrasensitive detection of miRNA-155 in breast cancer via direct hybridization assay using two-dimensional molybdenum disulfide field-effect transistor biosensor. Biosens Bioelectron 105:6–13. https://doi.org/10.1016/j.bios.2018.01.009 CrossRefGoogle Scholar
- 16.Deng H, Liu Q, Wang X, Huang R, Liu H, Lin Q, Zhou X, Xing D (2017) Quantum dots-labeled strip biosensor for rapid and sensitive detection of microRNA based on target-recycled nonenzymatic amplification strategy. Biosens Bioelectron 87:931–940. https://doi.org/10.1016/j.bios.2016.09.043 CrossRefPubMedGoogle Scholar
- 25.Hamd -Qaddare S, Salimi A (2017) Amplified fluorescent sensing of DNA using luminescent carbon dots and AuNPs/GO as a sensing platform: a novel coupling of FRET and DNA hybridization for homogeneous HIV-1 gene detection at femtomolar level. Biosens Bioelectron 89:773–780. https://doi.org/10.1016/j.bios.2016.10.033 CrossRefGoogle Scholar
- 27.Teymourian H, Salimi A, Khezrian S (2017) Development of a new label-free, indicator-free strategy toward ultrasensitive electrochemical DNA biosensing based on Fe3O4 anoparticles/ reduced graphene oxide composite. Electroanalysis 29:409–414. https://doi.org/10.1002/elan.201600336 CrossRefGoogle Scholar
- 29.Hamd-Ghadareh S, Salimi A, Fathi F, Bahrami S (2017) An amplified comparative fluorescence resonance energy transfer immunosensing of CA125 tumor marker and ovarian cancer cells using green and economic carbon dots for bio-applications in labeling, imaging and sensing. Biosens Bioelectron 96:308–316. https://doi.org/10.1016/j.bios.2017.05.003 CrossRefPubMedGoogle Scholar
- 30.Hamd-Ghadareh S, Salimi A, Parsa S, Fathi F (2018) Simultaneous biosensing of CA125 and CA15-3 tumor markers and imaging of OVCAR-3 and MCF-7 cells lines via bi-color FRET phenomenon using dual blue-green luminescent carbon dots with single excitation wavelength. Int J Biol Macromol 118:617–628. https://doi.org/10.1016/j.ijbiomac.2018.06.116 CrossRefPubMedGoogle Scholar