DNA-based one-pot rapid biosensing platform integrating turn-back loop primer-accelerated LAMP and Pyrococcus...

Area of Science:

• Food Science
• Biotechnology
• Analytical Chemistry

Background:

• Meat adulteration poses a global food safety risk.
• Existing detection methods can be time-consuming and require specialized facilities.
• There is a need for rapid, on-site, and reliable detection technologies.

Purpose of the Study:

• To develop an intelligent one-pot biosensing platform for rapid meat adulteration detection.
• To integrate TLAMP and PfAgo for sensitive and specific DNA detection.
• To create a cost-effective and user-friendly assay for point-of-care applications.

Main Methods:

• Development of the one-pot TLAMP-PfAgo assay (OTPA) integrating TLAMP and PfAgo.
• Utilizing a heat-activatable design with microcrystalline wax for reaction containment.
• Validation using commercial meat products and comparison with PCR methods.

Main Results:

• The OTPA assay achieved limits of detection as low as 3×10⁻⁴ ng/μL (pork DNA) and 2×10⁻⁴ ng/μL (beef DNA) within 30 minutes.
• Demonstrated sensitive and specific duplex target identification.
• Showed perfect concordance with standard PCR qualitative detection in commercial samples.
• Results were directly visualized under blue light.

Conclusions:

• The OTPA assay is a sensitive, specific, and rapid platform for detecting meat adulteration.
• The heat-activatable one-pot design ensures contamination-free and streamlined operation.
• This cost-effective and simple biosensing platform has significant potential for point-of-care testing and food safety surveillance.