Rapid and selective detection of macrocyclic trichothecene producing Stachybotrys chartarum strains by loop-mediated isothermal amplification (LAMP)

Anal Bioanal Chem. 2021 Aug;413(19):4801-4813. doi: 10.1007/s00216-021-03436-y. Epub 2021 Jun 15.

Abstract

Cytotoxic macrocyclic trichothecenes such as satratoxins are produced by chemotype S strains of Stachybotrys chartarum. Diseases such as stachybotryotoxicosis in animals and the sick building syndrome as a multifactorial disease complex in humans have been associated with this mold and its toxins. Less toxic non-chemotype S strains of S. chartarum are morphologically indistinguishable from chemotype S strains, which results in uncertainties in hazard characterization of isolates. To selectively identify macrocyclic trichothecene producing S. chartarum isolates, a set of sat14 gene-specific primers was designed and applied in a loop-mediated isothermal amplification (LAMP) assay using neutral red for visual signal detection. The assay was highly specific for S. chartarum strains of the macrocyclic trichothecene producing chemotype and showed no cross-reaction with non-macrocyclic trichothecene producing S. chartarum strains or 152 strains of 131 other fungal species. The assay's detection limit was 0.635 pg/rxn (picogram per reaction) with a reaction time of 60 min. Its high specificity and sensitivity as well as the cost-saving properties make the new assay an interesting and powerful diagnostic tool for easy and rapid testing.

Keywords: Diagnostic test kit; Indoor air quality; Sick building syndrome; Stachybotryotoxicosis; Water damage; sat14 gene.

MeSH terms

  • Genotype*
  • Macrocyclic Compounds / chemistry
  • Macrocyclic Compounds / metabolism*
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / methods*
  • Sensitivity and Specificity
  • Stachybotrys / genetics*
  • Stachybotrys / metabolism*
  • Trichothecenes / chemistry
  • Trichothecenes / metabolism*

Substances

  • Macrocyclic Compounds
  • Trichothecenes

Supplementary concepts

  • LAMP assay
  • Stachybotrys chartarum