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研究以戊糖片球菌16srDNA为靶目标设计引物,采用基因组DNA标准品以及质粒标准品构建两条标准曲线,建立了SYBR Green I荧光定量PCR法定量检测啤酒酿造过程中的戊糖片球菌的方法使用基因组DNA标准品构建标准曲线能去除DNA提取过程中损失对检测结果的影响,定量结果较质粒构建标准曲线的方法更为准确,标准曲线的相关系数0.994,检出限为10~3个/mL,得出的结果与平板计数法得出的结果无显著性差异,荧光定量PCR检测戊糖片球菌具有快速、灵敏的特点,适用于啤酒酿造过程中戊糖片球菌污染的检测。
In this study, primers were designed based on 16S rDNA of Pediococcus pentosaceus. Two standard curves were constructed using genomic DNA standards and plasmid standards. SYBR Green I quantitative PCR was used to quantitatively detect Pediococcus pentosaceus during beer brewing The construction of standard curve using the standard of genomic DNA can remove the influence of the loss during the DNA extraction process on the detection results. The quantitative results are more accurate than the standard curve of the plasmid. The correlation coefficient of the standard curve is 0.994 and the detection limit is 10-3 / mL. The results obtained by plate counting method showed no significant difference. The detection of Pediococcus pentosaceus by fluorescence quantitative PCR was rapid and sensitive, and was suitable for the detection of Pediococcus pentosaceus contamination in beer brewing.