基于流式细胞术的保加利亚乳杆菌活性检测方法应用研究

Application Research of Lactobacillus activity detection method based on flow cytometry

中文关键词:  流式细胞术  荧光染色  活性  保加利亚乳杆菌

英文关键词:flow cytometry  fluorescent staining  activity  Lactobacillus

中文摘要:

      【背景】对于发酵乳、饮料中的乳酸菌数量，国家的规范性文件GB 19302-2010《食品安全国家标准 发酵乳》对乳酸菌数做了明确的限量，即乳酸菌数要≥106CFU/mL(g)，以此来控制其产品质量。其中活的乳酸菌的活性更强，在人体消化系统环境中生存能力更强，人体在摄入后，到达肠道的数量会更多。【目的】基于流式细胞术的保加利亚乳杆菌活性检测方法，对现今售卖模式下的含保加利亚乳杆菌饮料中活性菌进行实时监测。【方法】 利用SYTO?24与PI双染色法对发酵乳饮料中的保加利亚乳杆菌的活性检测，并模拟现有售卖模式下发酵乳制品在其验货或销售等常温保存阶段保加利亚乳杆菌活性的变化进行监测。【结果】 在商超验货等常温放置时间在五个小时内，活性乳酸菌的数量级不会有显著性变化。【结论】 流式细胞术法能够对保加利亚乳杆菌活性有效检测，且能够单样品检测时间短，适合商品的快速检测和放行。由此可见，该方法对于需要快速放行的液体产品具有广阔的应用前景。

英文摘要:

      [Background] For fermented milk products, the state issued GB 19302-2010 "Food Safety National Standard Fermented Milk" to control its quality and safety, and made a clear limit on the number of lactic acid bacteria, that is, the number of lactic acid bacteria should be ≥10₆ CFU/mL (g). Among them, active lactic acid bacteria are more active, and their acid and alkali resistance is stronger. After ingesting the human body, the number of intestinal tracts will be more. [Objective] To establish a method for detecting the activity of lactic acid bacteria based on flow cytometry, and to simulate the change of lactic acid bacteria activity in fermented dairy products during the inspection and sale. [Methods] SYTO?24 and PI double staining were used to detect the activity of lactic acid bacteria in fermented milk beverages, and the results were compared with the plate counting method. [Results] The active bacterial community of lactic acid bacteria determined by this method has the ability to grow, and the number is not significantly different from the plate method. It can be used for the detection of lactic acid bacteria activity. The amount of lactic acid bacteria active bacteria will not change by an order of magnitude within five hours of normal temperature inspection. [Conclusion] Flow cytometry can effectively detect the activity of lactic acid bacteria, and the detection speed is fast. The single sample detection time is short and the sensitivity is high. It can be seen that the method has broad application prospects for liquid products requiring rapid release.