Abstract
Authenticity identification is crucial for the legal control of meat fraud and adulteration. Authentication capability of a method in mixed meat is essential for authenticity identification. This study aimed to develop a triplex real-time PCR method with better authentication capability to identify ovine, porcine, bovine, and equine DNAs in meat. The target DNAs were identified using specific primers and probes in the developed triplex real-time PCR technique, and an endogenous control was simultaneously amplified to eliminate false-negative results. The method had limits of detection (LOD) of 0.01–0.001 ng, 0.01–0.0005 ng, 0.1–0.001 ng, and 0.001–0.00005 ng for ovine, porcine, bovine, and equine, respectively. Furthermore, increasing the primer specificity enhanced this method’s authentication capability for complex meat from 10 to 0.1–1%. Overall, the results indicate that conservation for forward primer and specificity for reverse primer guarantee the compatibility and specificity of multiplex real-time PCR to detect minor adulteration. Therefore, this optimized method is an economical and efficient technique for authentication in food.
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This research was funded by the Science and Technology Program of Xilingol, grant numbers 202103 and BG202101.
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Liang Guo declares that he has no conflict of interest. Xiao Hai declares that he has no conflict of interest. Guo-Qiang Liu declares that he has no conflict of interest. Jian-Xing Luo declares that he has no conflict of interest. Yuan-Sheng Guo declares that he has no conflict of interest.
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Guo, L., Hai, X., Liu, GQ. et al. Enhancing the Authentication Capability of Triplex Real-Time PCR by Increasing the Primer Specificity. Food Anal. Methods 15, 2642–2651 (2022). https://doi.org/10.1007/s12161-022-02321-3
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DOI: https://doi.org/10.1007/s12161-022-02321-3