A label-free, PCR-free and signal-on electrochemical DNA biosensor for Leishmania major based on gold nanoleaves

Talanta. 2016 Dec 1:161:48-53. doi: 10.1016/j.talanta.2016.08.030. Epub 2016 Aug 9.

Abstract

Detection of leishmaniasis is important in clinical diagnoses. In the present study, identification of Leishmania parasites was performed by a label-free, PCR-free and signal-on ultrasensitive electrochemical DNA biosensor. Gold nanoleaves were firstly electrodeposited by an electrodeposition method using spermidine as a shape directing agent. The biosensor was fabricated by immobilization of a Leishmania major specific DNA probe onto gold nanoleaves, and methylene blue was employed as a marker. Hybridization of the complementary single stranded DNA sequence with the biosensor under the selected conditions was then investigated. The biosensor could detect a synthetic DNA target in a range of 1.0×10-10 to 1.0×10-19molL-1 with a limit of detection of 1.8×10-20molL-1, and genomic DNA in a range of 0.5-20ngμL-1 with a limit of detection of 0.07ngμL-1. The biosensor could distinguish Leishmania major from a non-complementary-sequence oligonucleotide and the tropica species with a high selectivity. The biosensor was applicable to detect Leishmania major in patient samples.

Keywords: DNA biosensor; Genosensor; Gold nanoleaves; Leishmaniasis.

MeSH terms

  • Biosensing Techniques*
  • DNA Probes / chemistry
  • DNA, Protozoan / analysis*
  • DNA, Protozoan / chemistry
  • DNA, Single-Stranded / analysis
  • DNA, Single-Stranded / chemistry
  • Electrochemical Techniques
  • Gold / chemistry
  • Humans
  • Immobilized Nucleic Acids / chemistry
  • Leishmania major / genetics*
  • Leishmaniasis, Cutaneous / microbiology*
  • Microscopy, Electron, Scanning
  • Nanostructures / chemistry
  • Nanostructures / ultrastructure

Substances

  • DNA Probes
  • DNA, Protozoan
  • DNA, Single-Stranded
  • Immobilized Nucleic Acids
  • Gold