Carbon nanodot-based electrogenerated chemiluminescence biosensor for miRNA-21 detection

Mikrochim Acta. 2021 Oct 30;188(11):398. doi: 10.1007/s00604-021-05038-y.

Abstract

A simple carbon nanodot-based electrogenerated chemiluminescence biosensor is described for sensitive and selective detection of microRNA-21 (miRNA-21), a biomarker of several pathologies including cardiovascular diseases (CVDs). The photoluminescent carbon nanodots (CNDs) were obtained using a new synthesis method, simply by treating tiger nut milk in a microwave reactor. The synthesis is environmentally friendly, simple, and efficient. The optical properties and morphological characteristics of the CNDs were exhaustively investigated, confirming that they have oxygen and nitrogen functional groups on their surfaces and exhibit excitation-dependent fluorescence emission, as well as photostability. They act as co-reactant agents in the anodic electrochemiluminescence (ECL) of [Ru(bpy)3]2+, producing different signals for the probe (single-stranded DNA) and the hybridized target (double-stranded DNA). These results paved the way for the development of a sensitive ECL biosensor for the detection of miRNA-21. This was developed by immobilization of a thiolated oligonucleotide, fully complementary to the miRNA-21 sequence, on the disposable gold electrode. The target miRNA-21 was hybridized with the probe on the electrode surface, and the hybridization was detected by the enhancement of the [Ru(bpy)3]2+/DNA ECL signal using CNDs. The biosensor shows a linear response to miRNA-21 concentration up to 100.0 pM with a detection limit of 0.721 fM. The method does not require complex labeling steps, and has a rapid response. It was successfully used to detect miRNA-21 directly in serum samples from heart failure patients without previous RNA extraction neither amplification process.

Keywords: Carbon nanomaterials; ECL biosensor; Green chemistry synthesis; RNA detection.

MeSH terms

  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Carbon / chemistry
  • Coordination Complexes / chemistry
  • Electrochemical Techniques / instrumentation
  • Electrochemical Techniques / methods*
  • Electrodes
  • Gold / chemistry
  • Heart Failure / blood
  • Humans
  • Immobilized Nucleic Acids / genetics
  • Limit of Detection
  • Luminescent Agents / chemistry*
  • Luminescent Measurements / instrumentation
  • Luminescent Measurements / methods*
  • Male
  • MicroRNAs / blood*
  • MicroRNAs / genetics
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides / chemistry
  • Oligodeoxyribonucleotides / genetics
  • Quantum Dots / chemistry*
  • Ruthenium Compounds / chemistry

Substances

  • Coordination Complexes
  • Immobilized Nucleic Acids
  • Luminescent Agents
  • MIRN21 microRNA, human
  • MicroRNAs
  • Oligodeoxyribonucleotides
  • Ruthenium Compounds
  • tris(2,2-bipyridine)-ruthenium(II)
  • Carbon
  • Gold