利用Real-time PCR技术分离鉴定酸乳中的乳双歧杆菌(Bifidobacterium Animalis Subsp.Lactis)  

Isolation and Identification of Bifidobacterium Animalis Subsp. Lactis in Yogurt by Real-Time PCR

作　　者：周燕霞 王青龙 任岩 李树垚 李龙 冉令磊 史锦硕 蔡雪凤 ZHOU Yanxia;WANG Qinglong;REN Yan;LI Shuyao;LI Long;RAN Linglei;SHI Jinshuo;CAI Xuefeng(Beijing Institute of Food Inspection(Beijing Food Safety Monitoring and Risk Assessment Center),Beijing 100094,China)

机构地区：[1]北京市食品检验研究院(北京市食品安全监控和风险评估中心),北京100094

出　　处：《食品科技》2023年第1期16-21,28,共7页Food Science and Technology

基　　金：国家市场监督管理总局科技计划项目(2019MK004)。

摘　　要：目的:利用Real-time PCR建立酸乳中乳双歧杆菌的快速精准鉴定方法。方法:选取乳双歧杆菌atpD基因,利用Primer Express 3.0.1设计引物探针,进而验证引物探针的特异性、灵敏度和抗干扰能力,最后应用到17种酸乳样品的检测。结果:所设计的引物探针可以有效地扩增出乳双歧杆菌,而对于20种乳酸菌和18种常见的致病菌都不能扩增,具有良好的特异性;其灵敏度在DNA水平可以达到1 pg/μL,在菌浓度水平可以达到10^(3)cfu/mL;并且不受酸乳中的嗜热链球菌和保加利亚乳杆菌的干扰,具有很好的抗干扰能力。利用RT-PCR技术能很好地鉴定17种酸乳中乳双歧杆菌。结论:该研究建立了酸乳中快速精准鉴定乳双歧杆菌的方法,对食品中双歧杆菌鉴定起到积极推动作用。Objective: A rapid and accurate identification method for Bifidobacterium animalis subsp. lactis in yoghurt by Real-time PCR was established. Methods: The atpD gene of Bifidobacterium animalis subsp. lactis was selected to be designed primer and probe using Primer Express 3.0.1 software, and the primer and probe were verified the specificity, sensitivity and anti-interference ability, finally the primer and probe were applied to detect the Bifidobacterium animalis subsp. lactis in 17 kinds of yogurt samples.Results: The primer and probe could effectively amplify Bifidobacterium animalis subsp. lactis except for 20 lactic acid bacteria and 18 common pathogenic bacteria, which had good specificity. The sensitivity of primer and probe could reach 1 pg/μL at the DNA level and 10^(3) cfu/mL at the bacterial concentration level, and was not interfered by Streptococcus thermophilus and Lactobacillus bulgaricus in yogurt, so it had good anti-interference ability. Bifidobacterium animalis subsp. lactis could be well identified in 17 kinds of yogurt by RT-PCR technology. Conclusion: This study establishes a method for rapid and accurate identification of Bifidobacterium animalis subsp. lactis in yogurt, which play a positive role in the identification of lactic acid bacteria in food.

关 键 词：乳双歧杆菌 实时荧光PCR 特异性 灵敏度 抗干扰性 

分 类 号：TS207.4[轻工技术与工程—食品科学]