Rapid and Simple Detection of Trichosporon asahii by Optimized Colony PCR

Area of Science:

• Medical Mycology
• Molecular Diagnostics
• Infectious Diseases

Background:

• Invasive trichosporonosis is primarily caused by *Trichosporon asahii*.
• Current detection methods for *T. asahii* are slow, expensive, and require complex sample preparation, delaying clinical diagnosis.
• Rapid and accurate identification of *T. asahii* is crucial for effective patient management.

Purpose of the Study:

• To evaluate colony PCR as a rapid diagnostic tool for *Trichosporon asahii*.
• To compare different DNA extraction methods for colony PCR suitability.
• To develop and validate species-specific primers for sensitive and specific *T. asahii* detection.

Main Methods:

• Colony PCR was assessed using four DNA extraction strategies: direct colony, freeze-thaw, glass beads, and enzymolysis.
• Species-specific primers targeting the IGS1 region of *T. asahii* ribosomal DNA were designed and screened.
• The optimized colony PCR method was used to detect *T. asahii* in mock clinical samples.

Main Results:

• The glass beads DNA extraction method combined with species-specific colony PCR demonstrated optimal performance.
• This method achieved a short procedure time of approximately 155 minutes.
• High sensitivity (detection limit of 10² CFU/mL) and specificity for *T. asahii* were observed.

Conclusions:

• Species-specific colony PCR using the glass beads method provides a rapid, simple, and sensitive approach for *T. asahii* identification.
• This technique significantly improves upon conventional diagnostic methods, facilitating quicker clinical decisions.
• The developed method holds promise for routine use in diagnosing invasive trichosporonosis.