荧光定量PCR法检测发酵果汁中嗜酸乳杆菌  

Detection of Lactobacillus acidophilus in Fermented Juice by Fluorescence Quantitative PCR

作　　者：王虹军 陈芝娟 李晓静 WANG Hongjun;CHEN Zhijuan;LI Xiaojing(Hangzhou Bioer Technology Co.,Ltd.,Hangzhou,Zhejiang 310053,China;Zhengzhou Grain Wholesale Market Co.,Ltd.,Zhengzhou,He'nan 450000,China)

机构地区：[1]杭州博日科技股份有限公司,浙江杭州310053 [2]郑州粮食批发市场有限公司,河南郑州450000

出　　处：《农产品加工》2022年第6期59-61,65,共4页Farm Products Processing

摘　　要：发酵饮品中目标菌种的定量检测对其质量与安全有着重要的意义。为了将发酵果汁中的嗜酸乳杆菌准确定量,采用现阶段食品微生物检测中广泛运用的分子学方法——荧光定量PCR法进行测定。根据嗜酸乳杆菌基因保守区域的序列设计其特异性引物和探针,以基因组纯化试剂盒提取纯化目标菌株的DNA为模板,利用荧光定量PCR方法先对模拟样品实施检测并制作出标准曲线,得到的相关系数为0.992,线性关系良好,可依此进行未知样品检测。最终检测出未知样品即发酵果汁中嗜酸乳杆菌的含量为2.27×10^(8) CFU/mL,并与平板计数结果基本一致。由此表明,荧光定量PCR法可快速准确地对发酵果汁中嗜酸乳杆菌实现定量检测。The quantitative detection of target bacteria in fermented materials is of great significance to the quality and safety of fermented drinks.In order to accurately quantify Lactobacillus acidophilus in fermented apple juice,the molecular method,fluorescence quantitative PCR,which is widely used in the detection of food microorganisms at present,was used to accomplish the determination.Design of specific primers and probes based on the sequence of conserved region of Lactobacillus acidophilus gene,extraction and purification of DNA of target strain with genomic purification kit as template,using fluorescence quantitative PCR to determine the simulated sample and draw the standard curve.The correlation coefficient was 0.992,and the linear relationship was good,which could be used to determine unknown samples.Finally,the quantity of Lactobacillus acidophilus in the unknown sample,i.e.fermented apple juice,was determined to be 2.27×10^(8) CFU/mL,which was basically consistent with the result of plate counting.The result showed that the fluorescent quantitative PCR method could detect Lactobacillus acidophilus in fermented apple juice quickly and accurately.

关 键 词：嗜酸乳杆菌 荧光定量PCR 定量检测 发酵果汁 

分 类 号：TS261.4[轻工技术与工程—发酵工程]