Highly sensitive enzyme-free immunosorbent assay for porcine circovirus type 2 antibody using Au-Pt/SiO2 nanocomposites as labels

Biosens Bioelectron. 2016 Aug 15:82:177-84. doi: 10.1016/j.bios.2016.04.001. Epub 2016 Apr 6.

Abstract

Improving the performance of conventional enzyme-linked immunosorbent assay (ELISA) is of great importance to meet the demand of early clinical diagnosis of various diseases. Herein, we report a feasible enzyme-free immunosorbent assay (EFISA) system using antibody conjugated Au-Pt/SiO2 nanocomposites (APS NCs) as labels. In this system, Au-Pt/SiO2 nanospheres (APS NPs) were first synthesized by wet chemical method and exhibited intrinsic peroxidase and catalase-like activity with excellent water-solubility. Then APS NCs were utilized as labels to replace HRP conjugated antibody, and Fe3O4 magnetic beads (MBs) to entrap the analyte. To discuss the performance of EFISA system, Human IgG was served as a model analyte, and porcine circovirus type 2 (PCV2) serums as real samples. The system boosted the detection limit of HIgG to 75pgmL(-1) with a RSD below 5%, a 264-fold improvement as compared with conventional ELISA. This is the first time that APS NCs have been used and successfully optimized for the sensitive dilution detection of PCV2 antibody (5:10(7)) in ELISA. Besides, APS NCs have advantages related to low cost, easy preparation, good stability and tunable catalytic activity, which make them a potent enzyme mimetic candidate and may find potential applications in bioassays and clinical diagnostics.

Keywords: Au-Pt/SiO(2) nanocomposites; Enzyme-free immunosorbent assay; Fe(3)O(4) magnetic beads; Human IgG; Porcine circovirus type 2.

MeSH terms

  • Animals
  • Antibodies, Viral / analysis
  • Circoviridae Infections / diagnosis
  • Circoviridae Infections / veterinary*
  • Circovirus / isolation & purification*
  • Gold / chemistry
  • Humans
  • Immunoglobulin G / analysis
  • Immunosorbent Techniques*
  • Immunosorbents / chemistry
  • Limit of Detection
  • Magnetite Nanoparticles / chemistry
  • Magnetite Nanoparticles / ultrastructure
  • Nanocomposites / chemistry*
  • Platinum / chemistry
  • Silicon Dioxide / chemistry*
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / virology

Substances

  • Antibodies, Viral
  • Immunoglobulin G
  • Immunosorbents
  • Magnetite Nanoparticles
  • Platinum
  • Gold
  • Silicon Dioxide