Dual approach for the colorimetric determination of unamplified microRNAs by using citrate capped gold nanoparticles
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The authors describe a method for the colorimetric determination of unamplified microRNA. It is based on the use of citrate-capped gold nanoparticles (AuNPs) and, alternatively, a microRNA-probe hybrid or a magnetically extracted microRNA that serve as stabilizers against the salt-induced aggregation of AuNPs. The absorbance ratios A525/A625 of the reacted AuNP solutions were used to quantify the amount of microRNA. The assay works in the range of 5–25 pmol microRNA. The lower limit of detection (LOD) is 10 pmol. The performance of the method was tested by detection of microRNA-210-3p in totally extracted urinary microRNA from normal, benign, and bladder cancer subjects. The sensitivity and specificity for qualitative detection of urinary microRNA-210-3p using the assay are 74% and 88% respectively, which is consistent with real time PCR based assays. The assay was applied to the determination of specific microRNA by using its specific oligo targeter or following magnetic isolation of the desired microRNA. The method is simple, cost-efficient, has a short turn-around time and requires minimal equipment and personnel.
KeywordsMicroRNA-210-3p MicroRNA detection Bladder cancer AuNPs Magnetic nanoparticles Salt-induced aggregation Oligonucleotide adsorption Oligotargeter Streptavidin
This work was financially supported by the national grant: RSTDF 6635. Authors acknowledge Professor Mohamed Esmat, Prof of urology for provision of clinical samples and patient’s data. The National patent application number of this work is 1631/2017.
Compliance with ethical standards
The authors declare that they have no competing interests.
- 2.Krol J, Sobczak K, Wilczynska U, Drath M, Jasinska A, Kaczynska D, Krzyzosiak WJ (2004) Structural features of microRNA (miRNA) precursors and their relevance to miRNA biogenesis and small interfering RNA/short hairpin RNA design. J Biol Chem 279:42230–42239. https://doi.org/10.1074/jbc.M404931200 CrossRefGoogle Scholar
- 4.Ben-Dov IZ, Tan YC, Morozov P, Wilson PD, Rennert H, Blumenfeld JD, Tuschl T (2014) Urine microRNA as potential biomarkers of autosomal dominant polycystic kidney disease progression: description of miRNA profiles at baseline. PLoS One 9:e86856. https://doi.org/10.1371/journal.pone.0086856 CrossRefGoogle Scholar
- 24.Farkhari N, Abbasian S, Moshaii A, Nikkhah M (2016) Mechanism of adsorption of single and double stranded DNA on gold and silver nanoparticles: Investigating some important parameters in bio-sensing applications. Colloids Surf B: Biointerfaces 148:657–664. https://doi.org/10.1016/j.colsurfb.2016.09.022 CrossRefGoogle Scholar
- 36.Zeng K, Li H, Peng Y (2017) Gold nanoparticle enhanced surface plasmon resonance imaging of microRNA-155 using a functional nucleic acid-based amplification machine. Microchim Acta. https://doi.org/10.1007/s00604-017-2276-2