A New Protocol for the Detection of Sterigmatocystin-producing Aspergillus Section Versicolores Using a High Discrimination Polymerase

Biocontrol Sci. 2020;25(2):113-118. doi: 10.4265/bio.25.113.

Abstract

Aspergillus section Versicolores species, except Aspergillus sydowii, produce a carcinogenic mycotoxin sterigmatocystin (STC). Since these fungi are found in varied environmental milieu including indoor dust and food products, our aim was to develop a sensitive and convenient assay to detect STC producing fungal strains. We made use of a high discrimination DNA polymerase (HiDi DNA polymerase), for single nucleotide polymorphism (SNP)-based PCR amplification. Using specific primer pairs based on the SNPs between A. sydowii and other strains of Aspergillus section Versicolores, we succeeded in amplifying the genomic DNA all target strains except A. sydowii. These results confirm that the SNP-based PCR amplification technique, using a high discrimination DNA polymerase, was a reliable and robust screening method for target fungal strains.

Keywords: Aspergillus section Versicolores; High discrimination polymerase; SNP-based PCR amplification.

MeSH terms

  • Aspergillus / genetics*
  • Aspergillus / isolation & purification
  • Aspergillus / metabolism
  • Base Sequence
  • Calmodulin / genetics
  • Calmodulin / metabolism
  • Carcinogens / analysis
  • Carcinogens / metabolism
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / metabolism
  • Fungal Proteins / genetics*
  • Fungal Proteins / metabolism
  • Gene Expression Regulation, Fungal*
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Polymorphism, Single Nucleotide*
  • RNA Polymerase I / genetics
  • RNA Polymerase I / metabolism
  • Sequence Alignment
  • Sterigmatocystin / analysis
  • Sterigmatocystin / biosynthesis

Substances

  • Calmodulin
  • Carcinogens
  • DNA, Fungal
  • Fungal Proteins
  • Sterigmatocystin
  • RNA Polymerase I
  • DNA-Directed DNA Polymerase

Supplementary concepts

  • Aspergillus sydowii
  • Aspergillus versicolor