Microchimica Acta

, 185:236 | Cite as

Dual approach for the colorimetric determination of unamplified microRNAs by using citrate capped gold nanoparticles

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Abstract

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.

Graphical abstract

Schematic of the two detection schemes: In the first approach, matched microRNA hybridizes with its specific probe to stabilize gold nanoparticles (AuNPs) against salt induced aggregation and to leave the red color of the AuNPs unchanged. In the second one, microRNA extracted via magnetic nanoparticles (MNP) stabilizes AuNPs against aggregation.

Keywords

MicroRNA-210-3p MicroRNA detection Bladder cancer AuNPs Magnetic nanoparticles Salt-induced aggregation Oligonucleotide adsorption Oligotargeter Streptavidin 
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Notes

Acknowledgements

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.

Supplementary material

604_2018_2767_MOESM1_ESM.docx (826 kb)
ESM 1 (DOCX 826 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biochemistry Department, Faculty of PharmacyMisr University for Science and Technology (MUST)GizaEgypt
  2. 2.Faculty of BiotechnologyOctober University for Modern Sciences & ArtsCairoEgypt
  3. 3.Oncology Diagnostic Unit, Medical Biochemistry & Molecular Biology Department, Faculty of MedicineAin Shams UniversityCairoEgypt
  4. 4.Faculty of Medicine Ain Shams Research Institute (MASRI)CairoEgypt

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