Novel microarrays for simultaneous serodiagnosis of multiple antiviral antibodies

PLoS One. 2013 Dec 18;8(12):e81726. doi: 10.1371/journal.pone.0081726. eCollection 2013.

Abstract

We developed an automated diagnostic system for the detection of virus-specific immunoglobulin Gs (IgGs) that was based on a microarray platform. We compared efficacies of our automated system with conventional enzyme immunoassays (EIAs). Viruses were immobilized to microarrays using a radical cross-linking reaction that was induced by photo-irradiation. A new photoreactive polymer containing perfluorophenyl azide (PFPA) and poly(ethylene glycol) methacrylate was prepared and coated on plates. Inactivated measles, rubella, mumps, Varicella-Zoster and recombinant Epstein-Barr viruse antigen were added to coated plates, and irradiated with ultraviolet light to facilitate immobilization. Virus-specific IgGs in healthy human sera were assayed using these prepared microarrays and the results obtained compared with those from conventional EIAs. We observed high correlation (0.79-0.96) in the results between the automated microarray technique and EIAs. The microarray-based assay was more rapid, involved less reagents and sample, and was easier to conduct compared with conventional EIA techniques. The automated microarray system was further improved by introducing reagent storage reservoirs inside the chamber, thereby conserving the use of expensive reagents and antibodies. We considered the microarray format to be suitable for rapid and multiple serological diagnoses of viral diseases that could be developed further for clinical applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral / blood*
  • Azides
  • Herpesvirus 4, Human / immunology
  • Humans
  • Hydrocarbons, Fluorinated
  • Immunoenzyme Techniques
  • Immunoglobulin G / blood
  • Mumps virus / immunology
  • Oligonucleotide Array Sequence Analysis*
  • Rubella virus / immunology
  • Serologic Tests / methods*

Substances

  • Antibodies, Viral
  • Azides
  • Hydrocarbons, Fluorinated
  • Immunoglobulin G
  • perfluorophenyl azide

Grants and funding

This work was sopported by Japan Society for the Promotion of Science KAKENHI Grant Number 22220009. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.