A "signal on" protection-displacement-hybridization-based electrochemical hepatitis B virus gene sequence sensor with high sensitivity and peculiar adjustable specificity

Biosens Bioelectron. 2016 Aug 15:82:212-6. doi: 10.1016/j.bios.2016.04.014. Epub 2016 Apr 6.

Abstract

One "signal on" electrochemical sensing strategy was constructed for the detection of a specific hepatitis B virus (HBV) gene sequence based on the protection-displacement-hybridization-based (PDHB) signaling mechanism. This sensing system is composed of three probes, one capturing probe (CP) and one assistant probe (AP) which are co-immobilized on the Au electrode surface, and one 3-methylene blue (MB) modified signaling probe (SP) free in the detection solution. One duplex are formed between AP and SP with the target, a specific HBV gene sequence, hybridizing with CP. This structure can drive the MB labels close to the electrode surface, thereby producing a large detection current. Two electrochemical testing techniques, alternating current voltammetry (ACV) and cyclic voltammetry (CV), were used for characterizing the sensor. Under the optimized conditions, the proposed sensor exhibits a high sensitivity with the detection limit of ∼5fM for the target. When used for the discrimination of point mutation, the sensor also features an outstanding ability and its peculiar high adjustability.

Keywords: Electrochemical sensor; Hepatitis B virus gene sequence; Methylene blue; Point mutation discrimination.

MeSH terms

  • Biosensing Techniques / instrumentation
  • Electrochemical Techniques / instrumentation*
  • Electrodes
  • Gold / chemistry
  • Hepatitis B / virology*
  • Hepatitis B virus / genetics*
  • Hepatitis B virus / isolation & purification
  • Humans
  • Limit of Detection
  • Methylene Blue / chemistry
  • Nucleic Acid Hybridization
  • Point Mutation

Substances

  • Gold
  • Methylene Blue