Establishment of TaqMan Real-time PCR Method for Detection of Wild Rice Derived Ingredients
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Graphical Abstract
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Abstract
In this study, a TaqMan real-time PCR method for rapid detection of wild rice in foods was established. Specific primers and probes were designed according to conserved sequence of internal transcribed space (ITS) gene of Z. latifolia, Z. aquatica, Z. palustris and Z. texana to test the target gene fragment in the samples, and then species-specific detection, sensitivity detection and practical application detection were carried out. Results showed that this real-time PCR method had strong specificity, only showed specific amplification curves for wild rice genomic DNA, and there was no amplification curve for other grains, animal and plant materials. The limit of detection was 0.001 ng/μL genomic DNA or 0.01% (W/W) wild rice powder per reaction. The feasibility of the method was further verified by detecting 100 samples of commercial imported wild rice, broken wild rice, coarse grain and rice flour. The wild rice ingredients were detected in 80 commercial imported wild rice and 5 broken wild rice samples. This method has the characteristics of strong specificity, high sensitivity, rapidity and high sufficiency, and is suitable for rapid identification of wild rice ingredients.
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