Rapid Russula senecis identification assays using loop-mediated isothermal amplification based on real-time...

Area of Science:

• Mycology
• Molecular Biology
• Food Safety

Background:

• Mushroom poisoning is a growing global health concern.
• Russula senecis is a common poisonous mushroom found in China.
• Accurate and rapid detection methods are crucial for public health.

Purpose of the Study:

• To develop novel loop-mediated isothermal amplification (LAMP) assays for detecting Russula senecis.
• To optimize LAMP assays for speed and sensitivity.
• To validate the applicability of LAMP assays for real-world samples.

Main Methods:

• Development of real-time fluorescence and visual loop-mediated isothermal amplification (LAMP) assays.
• Optimization of visual LAMP reaction time.
• Testing sensitivity with varying concentrations of genomic DNA.
• Validation using processed (fried, digested) mushroom samples and mixtures.

Main Results:

• Developed sensitive real-time and visual LAMP assays for R. senecis detection.
• Achieved a detection limit of 3.2 pg of genomic DNA.
• Successfully detected R. senecis in processed samples and mixtures down to 1% concentration.
• Demonstrated comparable performance to real-time PCR.

Conclusions:

• The developed LAMP assays are highly sensitive and specific for R. senecis.
• The assays are applicable to clinical and forensic identification, even with processed food samples.
• LAMP offers a reliable alternative to real-time PCR for mushroom poisoning detection.