Applying nanopore sequencing technology in Paracoccidioides sp.: a high-quality DNA isolation method for...

Area of Science:

• Mycology
• Genomics
• Infectious Diseases

Background:

• Paracoccidioidomycosis is a significant endemic mycosis in Latin America caused by Paracoccidioides fungi.
• Limited progress in Paracoccidioides genomics is attributed to incomplete reference databases and challenges in DNA extraction.
• Next-generation sequencing (NGS) offers potential for epidemiological surveillance and genomic characterization.

Purpose of the Study:

• To develop an improved DNA extraction method for Paracoccidioides species.
• To generate a high-coverage de novo genome assembly using Oxford Nanopore Technology (ONT).
• To overcome DNA extraction barriers hindering genomic studies of Paracoccidioides.

Main Methods:

• An improved DNA extraction protocol was developed specifically for Paracoccidioides.
• High-quality DNA was extracted and sequenced using Oxford Nanopore Technology (ONT).
• De novo genome assembly was performed using the generated long-read sequencing data.

Main Results:

• The study successfully generated a high-coverage de novo genome assembly for Paracoccidioides.
• The size of the obtained assembly was comparable to those generated using Illumina technology.
• This represents the first large-scale genome assembly of Paracoccidioides generated with ONT.

Conclusions:

• The developed DNA extraction method is effective for obtaining sufficient high-quality DNA for ONT sequencing.
• This advancement enables robust genomic characterization and epidemiological surveillance of Paracoccidioides.
• The study provides a valuable resource for future research into Paracoccidioidomycosis.