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A New Protocol for the Detection of Sterigmatocystin-producing Aspergillus Section Versicolores Using a High
Atsutaka Kubosaki1, Naoki Kobayashi2, Maiko Watanabe1
1Division of Microbiology, National Institute of Health Sciences.
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.
Insights
A new assay detects sterigmatocystin (STC)-producing fungi, excluding Aspergillus sydowii. This method uses single nucleotide polymorphism (SNP)-based PCR to reliably screen for these potentially carcinogenic strains in various environments.
Area of Science:
- Mycology
- Food Safety
- Molecular Biology
Background:
- Certain species within Aspergillus section Versicolores produce sterigmatocystin (STC), a carcinogenic mycotoxin.
- These fungi are frequently found in diverse environments, including indoor dust and food items, posing potential health risks.
- A need exists for a sensitive and convenient method to identify STC-producing fungal strains.
Purpose of the Study:
- To develop a sensitive and convenient assay for detecting sterigmatocystin (STC)-producing fungal strains.
- To differentiate STC-producing species within Aspergillus section Versicolores from non-producing species like Aspergillus sydowii.
Main Methods:
- Utilized high-discrimination DNA polymerase (HiDi DNA polymerase) for polymerase chain reaction (PCR) amplification.
- Employed single nucleotide polymorphism (SNP)-based PCR with specific primer pairs designed to target variations between Aspergillus sydowii and other Aspergillus section Versicolores strains.
- Successfully amplified genomic DNA from target strains.
Main Results:
- The SNP-based PCR assay effectively amplified genomic DNA from all targeted Aspergillus section Versicolores strains, with the exception of Aspergillus sydowii.
- This demonstrates the specificity of the primer pairs in distinguishing between STC-producing and non-STC-producing strains within the section.
Conclusions:
- The developed SNP-based PCR amplification technique, utilizing high-discrimination DNA polymerase, is a reliable and robust screening method.
- This assay can efficiently identify STC-producing fungal strains, contributing to improved food safety and environmental monitoring.
- The method offers a convenient approach for the detection of potentially carcinogenic fungi.
