A chemiluminescent immunoassay for the diagnosis of grapevine closteroviruses on nitrocellulose membrane

J Virol Methods. 1993 Apr;42(1):107-16. doi: 10.1016/0166-0934(93)90182-q.

Abstract

A dot-immunobinding assay was adapted on enhanced chemiluminescence (DIBA-ECL), which employs luminol, a cyclic diacylhydrazide, as substrate for horseradish peroxidase conjugated with a secondary antibody, for the diagnosis of grapevine closteroviruses I and III. The sensitivity of DIBA-ECL was also compared to other immunoenzymatic methods. DIBA-ECL proved to be at least 16 times more sensitive than the dot-immunobinding assay using chloronaphthol/diaminobenzidine mixture as a substrate, which was at least twice as sensitive as DAS-ELISA, DAS-indirect avidin-biotin complex ELISA, and dot-immunobinding assay, using alkaline phosphatase as enzyme. Optimisation of all parameters involved in the DIBA-ECL procedure and its advantages are discussed.

Publication types

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

MeSH terms

  • Collodion
  • Fruit / microbiology*
  • Immunoassay / methods*
  • Immunoblotting / methods*
  • Immunoenzyme Techniques
  • Luminescent Measurements
  • Membranes, Artificial
  • Plant Diseases
  • Plant Viruses / isolation & purification*
  • Sensitivity and Specificity

Substances

  • Membranes, Artificial
  • Collodion