Impedimetric genosensor for detection of hepatitis C virus (HCV1) DNA using viral probe on methylene blue doped silica nanoparticles

Int J Biol Macromol. 2017 May:98:84-93. doi: 10.1016/j.ijbiomac.2017.01.093. Epub 2017 Jan 23.

Abstract

An impedimetric genosensor was fabricated for detection of hepatitis C virus (HCV) genotype 1 in serum, based on hybridization of the probe with complementary target cDNA from sample. To achieve it, probe DNA complementary to HCVgene was immobilized on the surface of methylene blue (MB) doped silica nanoparticles MB@SiNPs) modified fluorine doped tin oxide (FTO) electrode. The synthesized MB@SiNPs was characterized using scanning electron microscopy (SEM), high resolution transmission electron microscopy (HRTEM) and X-ray diffraction (XRD) pattern. This modified electrode (ssDNA/MB@SiNPs/FTO) served both as a signal amplification platform (due to silica nanoparticles (SiNPs) as well as an electrochemical indicator (due to methylene blue (MB)) for the detection of the HCV DNA in patient serum sample. The genosensor was optimized and evaluated. The sensor showed a dynamic linear range 100-106 copies/mL, with a detection limit of 90 copies/mL. The sensor was applied for detection of HCV in sera of hepatitis patient and could be renewed. The half life of the sensor was 4 weeks. The MB@SiNPs/FTO electrode could be used for preparation of other gensensors also.

Keywords: Electrochemical impedance spectroscopy(EIS); Fluorine doped tin oxide(FTO) electrode; Hepatitis C; Hepatitis C virus (HCV); Impedimetric genosensor; Methylene blue; Probe DNA; Silica nanoparticles; cDNA.

MeSH terms

  • Base Sequence
  • Biosensing Techniques / instrumentation
  • DNA Probes / chemistry*
  • DNA Probes / genetics
  • DNA, Viral / analysis*
  • DNA, Viral / chemistry
  • DNA, Viral / genetics
  • Electric Impedance
  • Electrochemistry
  • Electrodes
  • Hepacivirus / genetics*
  • Humans
  • Limit of Detection
  • Methylene Blue / chemistry*
  • Nanocomposites / chemistry
  • Nanoparticles / chemistry*
  • Nucleic Acid Hybridization
  • Silicon Dioxide / chemistry*

Substances

  • DNA Probes
  • DNA, Viral
  • Silicon Dioxide
  • Methylene Blue