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。
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.
作者
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
CAO Weihua;LUO Na;SUN Yixuan;TU Jingxia;LIU Jing;WANG Deliang;HAO Jianqin;LUAN Chunguang;BAO Yihong(School of Forestry,Northeast Forestry University,Harbin 150040,China;China National Research Institute of,Beijing 100015,China;Guangzhou Nansha Zhujiang Brewery Co.Ltd,Guangzhou 511462,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2019年第21期53-59,共7页
Food and Fermentation Industries
基金
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)
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
Lactobacillus acetotolerans
RT-qPCR
specific primers
qualitative and quantitative detection
standard operating procdure(SOP)
