基于实时荧光定量环介导等温扩增技术快速检测发酵乳中嗜热链球菌和保加利亚乳杆菌
Development of Quantitative Loop-Mediated Isothermal Amplification for Rapid Detection of Streptococcus thermophilus and Lactobacillus bulgaricus in Fermented Milk
周帅康 1胡连霞 2王孟阳 3李苗 4卢涵 1王世杰5
作者信息
- 1. 河北科技大学食品与生物学院,河北 石家庄 050091
- 2. 石家庄学院化工学院,河北 石家庄 050035
- 3. 沧州市食品药品检验所,河北 沧州 060012
- 4. 君乐宝乳业集团股份有限公司,河北 石家庄 050221
- 5. 河北科技大学食品与生物学院,河北 石家庄 050091;君乐宝乳业集团股份有限公司,河北 石家庄 050221
- 折叠
摘要
基于实时荧光定量环介导等温扩增(quantitative loop-mediated isothermal amplification,qLAMP)技术建立发酵乳中嗜热链球菌和保加利亚乳杆菌活菌数快速定量方法.通过优化嗜热链球菌和保加利亚乳杆菌叠氮溴化丙锭(propidium monoazide,PMA)处理条件,对菌浓度对数与相应的循环阈值进行线性拟合,构建标准曲线,建立PMA-qLAMP检测方法.结果表明,该方法对嗜热链球菌的最低定量限为7.2×103 CFU/g,对保加利亚乳杆菌的最低定量限为5.6×104 CFU/g,将该方法应用于市售发酵乳产品活菌数定量检测,显示出良好的灵敏度和实用性.
Abstract
In this study,a rapid and quantitative method for the detection of viable bacterial counts of Streptococcus thermophilus and Lactobacillus bulgaricus in fermented milk was established using propidium monoazide(PMA)combined with quantitative loop-mediated isothermal amplification(PMA-qLAMP).The PMA treatment conditions for S.thermophilus and L.bulgaricus were optimized.A linear relationship was fitted between the logarithm of bacterial concentration and cycle threshold(Ct)value.The limit of quantification for S.thermophilus and L.bulgaricus was 7.2×103 and 5.6×104 CFU/g,respectively.The PMA-qLAMP method was applied to measure viable bacterial counts in commercial fermented milk,demonstrating good sensitivity and practicality.
关键词
发酵乳/环介导等温扩增/叠氮溴化丙锭/乳酸菌Key words
fermented milk/loop-mediated isothermal amplification/propidium monoazide/lactic acid bacteria引用本文复制引用
基金项目
中央引导地方科技发展资金项目(科技创新基地项目)(236Z2804G)
河北省"三三三人才工程"重点项目(A20231002)
河北省创新能力提升计划项目(22567685H)
石家庄市产业技术研究院建设项目(228790859A)
出版年
2024
国家科技期刊平台
NSTL
万方数据