Establishing a Universal Fluorescence Real-time PCR Method for the Detection of Yeast in Food
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Abstract:
The aim of this study was to establish a rapid fluorescence real-time PCR method for the detection of contamination in yeast present in food products. For this purpose, a universal probe and primers based on the yeast genome sequence were designed. The reaction system and conditions for fluorescence real-time PCR detection of yeast were established. The method was tested for its specificity, sensitivity, and reproducibility, and was also applied to the examination of food samples. The fluorescence real-time PCR assay was shown to display 100% specificity, as positive results for the specificity test were obtained only in yeast and not in bacteria or mold. The limit of detection of sensitivity was determined to be 760 cfu/mL. The sample was also tested for reproducibility, and the coefficient of variation (CV) was observed to fluctuate in the range of 0.62%~0.81% within the yeast group, and 0.43%~0.77% between groups. Moreover, a yeast enrichment test was performed, which demonstrated that the positive samples could be detected even after 12 h. In summary, the fluorescence real-time PCR assay established in this study showed excellent specificity, high sensitivity, and good reproducibility, in addition to being rapid and easy to use. This method could be used in the rapid detection of yeast contamination in food, and therefore can be used in research and related applications.
