马乳酒样乳杆菌马乳酒样亚种real-time PCR检测方法的建立与应用  

Establishment and Application of Real-Time PCR Detection Method for Lactobacillus kefiranofaciens subsp.kefiranofaciens

作　　者：吕厚姣 李欣媛 白小佳[1] 贾龙刚 耿伟涛 王艳萍[1] LÜHoujiao;LI Xinyuan;BAI Xiaojia;JIA Longgang;GENG Weitao;WANG Yanping(College of Food Science and Engineering,Tianjin University of Science and Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学食品科学与工程学院,天津300457

出　　处：《食品科学》2024年第9期102-108,共7页Food Science

摘　　要：本研究建立了一种特异性实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测方法,根据模式菌株马乳酒样乳杆菌马乳酒样亚种ZW3的16S rDNA序列和全基因组序列设计筛选特异性引物,采用SYBR Green I荧光染料建立real-time PCR方法,并对方法的特异性、灵敏度、重复性和混合体系等进行检测。结果表明,本研究所建立的方法特异性强、灵敏度高、重复性好,建立real-time PCR的标准曲线,其决定系数R2为0.965,具有良好的线性关系,且在马乳酒样乳杆菌马乳酒样亚种及混合体系中可以特异性检出。综上,本研究建立的real-time PCR法可以快速、准确地检测马乳酒样乳杆菌马乳酒样亚种,为马乳酒样乳杆菌的特异性定性定量检测提供了一种新的方法。This study developed a real-time polymerase chain reaction(real-time PCR)method for the detection of Lactobacillus kefiranofaciens subsp.kefiranofaciens.Based on the 16S rDNA gene sequence and whole genome sequence of the type strain of L.kefiranofaciens subsp.kefiranofaciens ZW3,specific primers were designed and screened.The fluorescent dye SYBR Green I was used in the real-time PCR method.Its specificity,sensitivity and repeatability were evaluated,and this method was applied to detect several strains of this species and its mixtures with other lactic acid bacteria.The results showed that the proposed method was highly specific,sensitive and repeatable.The standard curve was linear with a determination coefficient(R2)of 0.965.Moreover,this method could specifically detect L.kefiranofaciens subsp.kefiranofaciens from its mixture with other lactic acid bacteria.In summary,the real-time PCR method could quickly and accurately detect L.kefiranofaciens subsp.kefiranofaciens,providing a new method for the specific qualitative and quantitative detection of L.kefiranofaciens subsp.kefiranofaciens.

关 键 词：马乳酒样乳杆菌马乳酒样亚种 实时聚合酶链式反应 特异性引物 

分 类 号：Q93-331[生物学—微生物学]