Attomolar electrochemical detection of the BCR/ABL fusion gene based on an amplifying self-signal metal nanoparticle-conducting polymer hybrid composite

Colloids Surf B Biointerfaces. 2016 Dec 1:148:576-584. doi: 10.1016/j.colsurfb.2016.09.029. Epub 2016 Sep 22.

Abstract

In the last ten years, conjugated polymers started to be used in the immobilization of nucleic acids via non-covalent interactions. In the present study, we describe the construction and use of an electrochemical DNA biosensor based on a nanostructured polyaniline-gold composite, specifically developed for the detection of the BCR/ABL chimeric oncogene. This chromosome translocation is used as a biomarker to confirm the clinical diagnosis of both chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL). The working principle of the biosensor rests on measuring the conductivity resulting from the non-covalent interactions between the hybrid nanocomposite and the DNA probe. The nanostructured platform exhibits a large surface area that enhances the conductivity. Positive cases, which result from the hybridization between DNA probe and targeted gene, induce changes in the amperometric current and in the charge transfer resistance (RCT) responses. Atomic force microscopy (AFM) images showed changes in the genosensor surface after exposure to cDNA sample of patient with leukemia, evidencing the hybridization process. This new hybrid sensing-platform displayed high specificity and selectivity, and its detection limit is estimated to be as low as 69.4 aM. The biosensor showed excellent analytical performance for the detection of the BCR/ABL oncogene in clinical samples of patients with leukemia. Hence, this electrochemical sensor appears as a simple and attractive tool for the molecular diagnosis of the BCR/ABL oncogene even in early-stage cases of leukemia and for the monitoring of minimum levels of residual disease.

Keywords: BCR/ABL fusion gene; Biosensor; Electrochemistry; Hybrid nanoparticles; Leukemia.

MeSH terms

  • Aniline Compounds / chemistry
  • Biosensing Techniques / methods
  • DNA Probes / chemistry
  • DNA Probes / genetics
  • DNA, Neoplasm / chemistry
  • DNA, Neoplasm / genetics
  • Electric Conductivity
  • Electrochemical Techniques / methods
  • Fusion Proteins, bcr-abl / genetics*
  • Gold / chemistry
  • Humans
  • Leukemia / diagnosis
  • Leukemia / genetics
  • Metal Nanoparticles / chemistry*
  • Microscopy, Atomic Force
  • Nanocomposites / chemistry*
  • Polymerase Chain Reaction
  • Polymers / chemistry*
  • Reproducibility of Results
  • Sensitivity and Specificity

Substances

  • Aniline Compounds
  • DNA Probes
  • DNA, Neoplasm
  • Polymers
  • polyaniline
  • Gold
  • Fusion Proteins, bcr-abl