Development of a rapid on-site detection method for pork in processed meat products using real-time loop-mediated isothermal amplification

• LAMP primers were designed based on the D-loop gene in pork and the 18S rRNA gene.
• The method was able to detect pork within 30 min without DNA extraction.
• Forty-two processed meat products were analyzed using this direct, real-time LAMP method.
• This assay can be applied to the on-site identification of pork in processed meat.

On-site detection with minimal equipment and no risk of contamination of samples is crucial for the rapid and sensitive identification of pork in processed meat products. To resolve this issue, a reliable loop-mediated isothermal amplification (LAMP) method was developed to detect pork in meat using a portable real-time fluorometer without the need for DNA extraction. Pork-specific primers for the LAMP assay were designed based on the mitochondrial D-loop regions, and eukaryotic primers based on the 18S rRNA gene were used for the endogenous control. The adoption of an endogenous control prevented false-negative results. The detection level was 1 pg for raw pork DNA and 0.1% of pork in a beef-meat mixture. The applicability of the developed method was demonstrated in commercially processed meat products. Forty-two meat products were successfully verified for labeling compliance using this method within 30 min without the need for nucleic acid extraction.