Abstract
The rising price of sika deer meat is increasing the risk of economic adulteration, highlighting the need for rapid and reliable authentication methods to protect both market integrity and consumers. This work presents a novel countermeasure: a polymerase chain reaction (PCR)-based nucleic acid test strip designed for the specific and visual identification of sika deer meat. Our approach commenced with the design of specific primers targeting the cytochrome C oxidase subunit I (COI) gene. To guarantee the reliability of the assay, a DNA standard plasmid was constructed to serve as an unambiguous positive control for the PCR. Under optimized conditions, results showed that authentic sika deer meat generated both test and control lines on the strip, while adulterated and negative samples produced only the control line. The assay demonstrated flawless specificity and a detection sensitivity of 1.0 ng·μL(-1) for target DNA, representing a tenfold enhancement over gel electrophoresis. Furthermore, the method demonstrated a detection limit of 1% for sika deer meat in admixed samples, with a faint but visible signal observed down to 0.1% under optimized conditions. In conclusion, the developed test strip method is not only specific and sensitive but also user-friendly, positioning it as a practical and powerful tool for rapid, on-site meat authentication.