Development of a highly sensitive gold nanoparticle probe-based assay for bluetongue virus detection

J Virol Methods. 2012 Jul;183(1):45-8. doi: 10.1016/j.jviromet.2012.03.027. Epub 2012 Mar 30.

Abstract

A simple gold nanoparticle (GNP) probe based assay (GNPA) that was modified from a bio-barcode assay (BCA) technique, was developed for ultra-sensitive, rapid detection of the bluetongue virus (BTV) VP7 outer-core protein. This assay captures the VP7 target antigen using the GNP probe coated with anti-VP7 polyclonal antibodies and single-stranded signal DNA. Magnetic microparticle (MMP) probes coated with anti-VP7 monoclonal antibodies were then added to form a sandwich immuno-complex. The single-stranded signal DNA coated onto the GNP probe present in the immuno-complex could then be detected by PCR and real-time fluorescence PCR using a TaqMan probe. The assay has a purified VP7 detection limit of 10(-2)fg/ml which is 8 orders of magnitude greater than that of conventional antigen capture ELISAs and 1 order of magnitude more sensitive than that of a conventional BCA. These results indicate that the GNPA is a highly sensitive method for easy detection of BTV proteins and that it can be modified as needed to measure the presence of other proteins.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral
  • Antigens, Viral / analysis
  • Bluetongue / diagnosis*
  • Bluetongue / virology
  • Bluetongue virus / isolation & purification*
  • Clinical Laboratory Techniques / methods*
  • Immunoassay / methods
  • Nanoparticles*
  • Polymerase Chain Reaction / methods
  • Sensitivity and Specificity
  • Sheep
  • Veterinary Medicine / methods*
  • Viral Core Proteins / analysis*
  • Virology / methods*

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Viral Core Proteins
  • VP7 protein, orbivirus