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Rapid identification of Ochroconis gallopava by a loop-mediated isothermal amplification (LAMP) method
Akira Ohori1, Shigeo Endo, Ayako Sano
1Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, 1-8-1 Inohana, Chiba 260-8673, Japan.
Abstract:
Ochroconis gallopava is a species of dematiaceous fungi recognized as a causative agent of zoonotic and emerging fungal infections. It affects the central nervous system and respiratory tracts of humans, birds and cats. We designed O. gallopava species-specific primer sets to aid in its identification by a loop-mediated isothermal amplification (LAMP) method based on the D1/D2 domain of the LSU rDNA sequence. The LAMP method successfully detected the gene from both fungal DNA and experimentally infected brains and spleens of mice and will be helpful in the diagnosis of O. gallopava infection.
Insights
A new loop-mediated isothermal amplification (LAMP) method was developed for identifying Ochroconis gallopava, a fungus causing emerging infections. This diagnostic tool accurately detects the fungus in various samples, aiding in O. gallopava infection diagnosis.
Area of Science:
- Mycology
- Infectious Diseases
- Molecular Diagnostics
Background:
- Ochroconis gallopava is a dematiaceous fungus causing zoonotic and emerging infections.
- This fungus affects the central nervous system and respiratory tracts in humans, birds, and cats.
- Accurate and rapid identification is crucial for managing O. gallopava infections.
Purpose of the Study:
- To develop a species-specific identification method for Ochroconis gallopava.
- To utilize the loop-mediated isothermal amplification (LAMP) technique for fungal detection.
- To establish a diagnostic tool for O. gallopava infections.
Main Methods:
- Designed O. gallopava species-specific primer sets targeting the D1/D2 domain of the LSU rDNA sequence.
- Employed the loop-mediated isothermal amplification (LAMP) method for fungal DNA detection.
- Validated the LAMP assay using fungal DNA and experimentally infected mouse tissues (brains and spleens).
Main Results:
- The developed LAMP method demonstrated high specificity for O. gallopava.
- Successful detection of O. gallopava DNA from pure cultures and infected mouse tissues.
- The assay proved effective in identifying the fungus in complex biological samples.
Conclusions:
- The O. gallopava-specific LAMP assay provides a rapid and reliable method for fungal identification.
- This molecular tool can significantly aid in the clinical and veterinary diagnosis of O. gallopava infections.
- The LAMP method offers a valuable advancement in detecting emerging fungal pathogens.
