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High‐Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus

Summary

Scientists have developed a faster, cheaper way to grow and extract DNA from Aspergillus fumigatus, a fungus that causes serious infections in sick people. Using 96-well plates (the same format used in many laboratories), researchers can now process many samples at once instead of handling them one by one. The method works well for identifying drug-resistant fungal strains and preparing DNA for genetic analysis.

Background

Aspergillus fumigatus is an opportunistic human fungal pathogen widely distributed in the environment that causes serious diseases including invasive aspergillosis in immunocompromised patients. There is a critical need to screen large numbers of environmental and clinical isolates for phenotypic and genotypic characteristics, particularly to identify antifungal-resistant strains, but current protocols are time-consuming and labor-intensive.

Objective

The objective is to develop a novel high-throughput approach that enables researchers to culture individual A. fumigatus isolates and isolate DNA of sufficient quality for PCR and whole genome sequencing in a 96-well plate format. This method aims to significantly reduce cost, labor, and incubator space requirements compared to currently available protocols.

Results

The protocols successfully produced sufficient spore suspensions from mini slants for downstream applications and yielded PCR-quality DNA suitable for amplicon sequencing and genotyping. High-purity genomic DNA isolated from mycelial mats was of sufficient quality for whole genome sequencing, with >99% variant congruency to gold-standard methods at sequencing depths above 15×.

Conclusion

These novel high-throughput protocols enable efficient screening of large numbers of A. fumigatus isolates in 96-well format, reducing time, labor, and consumable costs while maintaining DNA quality for downstream molecular analysis including PCR, Sanger sequencing, and whole genome sequencing.
  • Published in:Current Protocols,
  • Study Type:Protocol Development Study,
  • Source: PMID: 40172288, DOI: 10.1002/cpz1.70112
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