Onsite rapid detection method for genetically modified maize and soybean based on recombinase polymerase...

Area of Science:

• Agricultural Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Commercial planting of transgenic crops in China necessitates rapid on-site detection methods.
• Existing methods for detecting genetically modified organisms (GMOs) often lack speed and portability.
• Ensuring the healthy development of China's biological breeding industry requires efficient GMO monitoring.

Purpose of the Study:

• To develop a rapid, on-site detection method for transgenic maize and soybeans.
• To overcome the limitations of current on-site GMO detection techniques.
• To provide technical support for the on-site detection of transgenic crops.

Main Methods:

• Utilized recombinase polymerase amplification (RPA) for sensitive and specific DNA amplification.
• Designed and screened specific primers and probes for target transgenic events.
• Developed rapid DNA extraction protocols for seed and leaf samples, including chlorophyll interference mitigation.
• Integrated portable equipment for amplification and visual detection.

Main Results:

• Established a highly specific and sensitive RPA system detecting down to 20 copies per transgenic event.
• Developed rapid DNA extraction from seeds using nucleic acid release agents.
• Created a method for leaf DNA extraction that eliminates chlorophyll interference.
• Constructed a complete on-site detection system yielding results consistent with conventional PCR in approximately 20 minutes.

Conclusions:

• The developed RPA-based method offers a practical and adaptable solution for on-site transgenic maize and soybean detection.
• The technique is suitable for field use, requiring minimal equipment.
• This approach provides valuable technical support for GMO monitoring and serves as a reference for other rapid nucleic acid detection technologies.