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。
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
. 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
.
DOI: 10.13995/j.cnki.11-1802/ts.022197
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.
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