Collection, nucleic acid release, amplification, and visualization platform for rapid field detection of rice false...

Area of Science:

• Agricultural Science
• Biotechnology
• Food Safety

Background:

• Rice false smut (RFS) poses significant global food safety risks.
• Current diagnostic methods like traditional polymerase chain reaction (PCR) are unsuitable for rapid field detection due to complex sample handling.
• There is a critical need for efficient, on-site diagnostic tools for RFS.

Purpose of the Study:

• To develop a simple, portable microfluidic platform for the rapid detection of RFS.
• To integrate multiple diagnostic steps onto a single chip for streamlined field analysis.
• To provide a timely and accurate tool for monitoring RFS in agricultural settings.

Main Methods:

• Development of a microfluidic chip integrating spore purification, nucleic acid release, and loop-mediated isothermal amplification (LAMP).
• Utilized a micro air pump for efficient spore separation and collection.
• Employed benzyl chloride method for rapid nucleic acid release and SYBR Green I for visual detection of LAMP products.

Main Results:

• Achieved approximately 78% spore collection efficiency on the microfluidic chip.
• Demonstrated 100% specificity and a detection limit of 100 copies/reaction.
• Exhibited high stability (CV < 5%), excellent quantitative ability (R² = 0.989), and reduced detection time by ~20 minutes compared to RT-PCR.

Conclusions:

• The developed microfluidic chip offers a rapid, portable, and efficient solution for RFS detection.
• The integrated platform simplifies operations by avoiding complex nucleic acid extraction and large instruments.
• This tool holds potential for real-time monitoring and early warning systems for RFS in rice paddies.