A highly specific competitive direct enzyme immunoassay for sterigmatocystin as a tool for rapid immunochemotaxonomic differentiation of mycotoxigenic Aspergillus species

Lett Appl Microbiol. 2017 Feb;64(2):124-130. doi: 10.1111/lam.12702.

Abstract

A simplified method to produce specific polyclonal rabbit antibodies against sterigmatocystin (STC) was established, using a STC-glycolic acid-ether derivative (STC-GE) conjugated to keyhole limpet haemocyanin (immunogen). The competitive direct enzyme immunoassay (EIA) established for STC had a detection limit (20% binding inhibition) of 130 pg ml-1 . The test was highly specific for STC, with minor cross-reactivity with O-methylsterigmatocystin (OMSTC, 0·87%) and negligible reactivity with aflatoxins (<0·02%). STC-EIA was used in combination with a previously developed specific EIA for aflatoxins (<0·1% cross-reactivity with STC and OMSTC), to study the STC/aflatoxin production profiles of reference strains of Aspergillus species. This immunochemotaxonomic procedure was found to be a convenient tool to identify STC- or aflatoxin-producing strains.

Significance and impact of the study: The carcinogenic mycotoxin sterigmatocystin (STC) is produced by several Aspergillus species, either alone or together with aflatoxins. Here, we report a very simple and straightforward procedure to obtain highly sensitive and specific anti-STC antibodies, and their use in the first ever real STC-specific competitive direct enzyme immunoassay (EIA). In combination with a previous EIA for aflatoxins, this study for the first time demonstrates the potential of a STC/aflatoxin EIA pair for what is branded as 'immunochemotaxonomic' identification of mycotoxigenic Aspergillus species. This new analytical tool enhances analytical possibilities for differential analysis of STC and aflatoxins.

Keywords: Aspergillus; aflatoxin; immunoassay; mycotoxin; polycolonal antibodies; sterigmatocystin.

Publication types

  • Evaluation Study

MeSH terms

  • Aflatoxins / analysis*
  • Aflatoxins / biosynthesis
  • Aflatoxins / immunology
  • Antibodies / immunology
  • Aspergillus / classification
  • Aspergillus / isolation & purification*
  • Aspergillus / metabolism
  • Cross Reactions / immunology
  • Immunoenzyme Techniques*
  • Sensitivity and Specificity
  • Sterigmatocystin / analogs & derivatives*
  • Sterigmatocystin / analysis
  • Sterigmatocystin / immunology
  • Sterigmatocystin / metabolism

Substances

  • Aflatoxins
  • Antibodies
  • Sterigmatocystin
  • O-methylsterigmatocystin