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食品与发酵工业  2019, Vol. 45 Issue (21): 53-59    DOI: 10.13995/j.cnki.11-1802/ts.022197
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
啤酒腐败菌耐乙酸乳杆菌种特异性检测体系的建立与应用
曹伟华1,2, 罗娜3, 孙义玄1,2, 涂京霞3, 刘静3, 王德良2, 郝建秦2, 栾春光2*, 包怡红1*
1 (东北林业大学 林学院,黑龙江 哈尔滨,150040)
2 (中国食品发酵工业研究院有限公司,北京,100015)
3 (广州南沙珠江啤酒有限公司,广东 广州,511462)
Establishment and application of a species-specific detection system for beerspoilage bacteria-Lactobacillus acetotolerans
CAO Weihua1,2, LUO Na3, SUN Yixuan1,2, TU Jingxia3, LIU Jing3, WANG Deliang2, HAO Jianqin2, LUAN Chunguang2*, BAO Yihong1*
1 (School of Forestry, Northeast Forestry University, Harbin 150040, China)
2 (China National Research Institute of Food and Fermentation Industries,Beijing 100015, China)
3 (Guangzhou Nansha Zhujiang Brewery Co.Ltd,Guangzhou 511462, China)
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摘要 以耐乙酸乳杆菌的3个特异性基因为靶标,设计并优化了3对耐乙酸乳杆菌的特异性引物,建立了基于SYBR GREEN实时荧光定量PCR的耐乙酸乳杆菌定性定量检测的标准检测(standard operation procedure, SOP)方法。利用耐乙酸乳杆菌CN247的基因组DNA作为标准品绘制了标准曲线,实现了对耐乙酸乳杆菌的定量检测,耐乙酸乳杆菌的最低检测限可达100 CFU/mL,精度可达10 CFU/mL。大生产样品在培养基中富集培养48 h后即可满足检测的要求,检出率为100%,整个检测时间控制在3 d以内,满足了啤酒企业对出厂产品快速检测的质量控制要求。
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曹伟华
罗娜
孙义玄
涂京霞
刘静
王德良
郝建秦
栾春光
包怡红
关键词:  耐乙酸乳杆菌  RT-qPCR  特异性引物  定性定量检测  标准检测方法    
Abstract: A species-specific PCR detection system was established for beer spoilage bacteria Lactobacillus acetotolerans CN247. Three sets of species-specific primers were designed and optimized in SYBR GREEN real-time PCR detection system. A standard operating procedure (SOP) for qualitative and quantitative detection of Lactobacillus acetotolerans in beer samples was obtained. The results showed that the detection limit for Lactobacillus acetotolerans reached 102 CFU/mL and the detection accuracy reached 101 CFU/mL by using the detection system developed here. The results could be obtained within 48 h after pre-enrichment treatment by using NBB media. This research developed a Lactobacillus acetotolerans species-specific detection system which could meet the quality control requirements of the manufacturer in beer industry.
Key words:  Lactobacillus acetotolerans    RT-qPCR    specific primers    qualitative and quantitative detection    standard operating procdure(SOP)
收稿日期:  2019-09-06                出版日期:  2019-11-15      发布日期:  2019-11-15      期的出版日期:  2019-11-15
基金资助: 广州市科技计划项目珠江科技新星专项(201610010183);北京市科技新星计划(Z181100006218004)
作者简介:  曹伟华硕士研究生和罗娜硕士研究生为共同第一作者(栾春光高级工程师和包怡红教授为共同通讯作者,E-mail:chunguangluan@163.com;baoyihong@163.com)。
引用本文:    
曹伟华,罗娜,孙义玄,等. 啤酒腐败菌耐乙酸乳杆菌种特异性检测体系的建立与应用[J]. 食品与发酵工业, 2019, 45(21): 53-59.
CAO Weihua,LUO Na,SUN Yixuan,et al. Establishment and application of a species-specific detection system for beerspoilage bacteria-Lactobacillus acetotolerans[J]. Food and Fermentation Industries, 2019, 45(21): 53-59.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.022197  或          http://sf1970.cnif.cn/CN/Y2019/V45/I21/53
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