食品科学

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深度测序技术检测益生菌产品菌株组成及16S rDNA序列

杨捷琳,袁辰刚,窦同海,丁小磊,潘良文   

  1. 1.上海出入境检验检疫局,上海 200135;2. 复旦大学生命科学学院,上海 200433
  • 出版日期:2013-10-25 发布日期:2013-09-28

Application of Next-Generation Sequencing (NGS) Technology in Analysis of Bacterial Species Composition and 16S rDNA Sequencing of Commercial Yoghurt Products

YANG Jie-lin,YUAN Chen-gang,DOU Tong-hai,DING Xiao-lei,PAN Liang-wen   

  1. 1. Shanghai Entry and Exit Inspection and Quarantine Bureau, Shanghai 200135, China;
    2. School of Life Sciences, Fudan University, Shanghai 200433, China
  • Online:2013-10-25 Published:2013-09-28

摘要:

目的:探查益生菌制品的标签标示与产品中实际检测到的菌株比较及不同产品中菌株的异同性。方法:以市场上购买的进口、出口酸乳制品作为研究对象,设计可区别24种产品的样品标签序列(Barcode),通过PCR及Illumina平台深度测序分析,获得了酸乳制品中所含益生菌种类及序列等详细信息,根据Barcode序列信息对其中所含的24个混合样本进行区分,然后利用16S RDP的16S数据库,对测序的Reads进行比对。结果:24种产品中标签标示与样品中实际检测到的菌株完全一致的仅有两种,在24种酸乳制品中,以嗜热链球菌、嗜酸乳杆菌、保加利亚乳杆菌/德式乳杆菌为主要发酵菌株,双歧杆菌作为添加的益生菌基本在所有样品中存在,但含量较低,另一方面,不同产品中同一菌株的16S rDNA序列相似度较高,仅有两种菌株的1~2个碱基存在差异。结论:深度测序是一种可快速有效对食品中微生物群体精确筛查的技术,能够精确深入了解食品中微生物群落。使用溶方法检测目前市场上的酸乳制品,可发现使用的发酵菌种多来自商业化购买的菌剂,同质化程度较高,而作为后添加的双歧杆菌由于其生长条件的苛刻,实际活菌数量较低。

关键词: 乳酸菌, 酸乳制品, 16S rDNA, 深度测序

Abstract:

In recent years, an increasing number of applications of probiotic dairy products have been reported to maintain
the intestinal flora balance, treat chronic and acute diarrhea and enhance the immune system. Nevertheless, many probiotic
products from the market are often not accurately labeled for the bacterial composition. Therefore, it is necessary to know
the bacterial composition of probiotic products for better judgment of their quality. For this purpose, we investigated the
accuracy and feasibility of 16S rDNA sequencing using Illumina next-generation sequencing (NGS) platform for bacterial
identification of the bacterial species composition in probiotic foods, especially in 24 probiotic products from 19 manufacturers.
Data showed that some probiotic products contained unadvertised lactobacilli and bifidobacteria, whereas others were
not the same as labeled. In summary, 16S rDNA sequencing by Illumina NGS platform is a rapid method for profiling the
microbial community in foods and clinical studies.

Key words: lactic acid bacterial, yoghurt, 16S rDNA, next-generation sequencing