An improved gold nanoparticle probe-based assay for HCV core antigen ultrasensitive detection

J Virol Methods. 2017 May:243:142-145. doi: 10.1016/j.jviromet.2017.02.007. Epub 2017 Feb 8.

Abstract

A gold nanoparticle probe-based assay (GNPA) was developed for ultrasensitive detection of Hepatitis C virus (HCV) core antigen. In the GNPA, after anti-HCV core antigen polyclonal antibodies and single-stranded barcode signal DNA were labeled on gold nanoparticle probe (NP), DNA enzyme was used to degrade the unbound barcode DNAs. The anti-HCV core antigen monoclonal antibodies were coated on magnetic microparticles probe (MMP). Then the NP-HCV core antigen-MMP sandwich immuno-complex was formed when the target antigen protein was added and captured. Magnetically separated, the immuno-complex containing the single-stranded barcode signal DNA was characterized by TaqMan probe based real-time fluorescence PCR. A detection limit of 1 fg/ml was determined for the HCV core antigen which is magnitude greater than that of ELISA (2ng/ml). The coefficients of variation (CV) of intra-assay and inter-assay respectively ranged from 0.22-2.62% and 1.92-3.01%. The improved GNPA decreased the interference of unbound barcode DNAs and may be an new way for HCV core antigen detection.

Keywords: Bio-bar code assay; Gold nanoparticle probe based assay; Hepatitis C virus; Ultrasensitive detection.

Publication types

  • Evaluation Study

MeSH terms

  • Antigens, Viral / analysis*
  • Gold*
  • Hepacivirus / isolation & purification*
  • Hepatitis C / diagnosis*
  • Humans
  • Immunoassay / methods*
  • Nanoparticles*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Viral Core Proteins / analysis*

Substances

  • Antigens, Viral
  • Viral Core Proteins
  • nucleocapsid protein, Hepatitis C virus
  • Gold