靶向鉴别、检测动物双歧杆菌的引物探针及应用  

Primer probes for targeted identification and detection of Bifidobacterium animalis and their application

作　　者：杜丽霞 李典 禚惠荣 石梦楠 陈延宁 李岚芳 侯少阳 DU Lixia;LI Dian;ZHUO Huirong;SHI Mengnan;CHEN Yanning;LI Lanfang;HOU Shaoyang(College of Pharmacy,Heze University,Heze 274015,China)

机构地区：[1]菏泽学院药学院,菏泽274015

出　　处：《中国药科大学学报》2023年第2期255-262,共8页Journal of China Pharmaceutical University

基　　金：山东省自然科学基金资助项目(No.ZR2022QB146);菏泽学院博士基金资助项目(No.XY21BS35)。

摘　　要：为解决目前应用于鉴别动物双歧杆菌的方法的用时长、操作复杂、实验环境低适配性等缺点,开发了一种基于ERIC-PCR技术设计特异性引物探针以靶向鉴别、检测动物双歧杆菌的方法。以动物双歧杆菌HP-B1124基因组DNA为基础,优化动物双歧杆菌HP-B1124的ERIC-PCR反应条件,对ERIC-PCR片段逐一回收并进行测序,针对测序结果设计两对特异性引物探针,对两对引物探针设计结果的准确性、特异性、可检测基因组DNA的含量限度、普适性进行检验,并运用所设计的两对特异性引物探针检验市售公示配方中含有动物双歧杆菌的产品。本研究设计的两对特异性引物探针可简单、快速、靶向的用于动物双歧杆菌的鉴定。此方法优化了目前相对传统的动物双歧杆菌纯培养及平板计数鉴定方法,一定程度上解决了SNP基因分型技术及实时荧光定量PCR法等方法在鉴定动物双歧杆菌方面对实验设备及试剂的高要求,具有成本低、特异性强的特点,有较为广阔的市场开发前景。In this study,in order to overcome the shortcomings of the current methods used to identify Bifidobac⁃terium animalis,such as long time,complicated operation and low adaptability of experimental environment,specific primer probes were designed based on ERIC-PCR technology to identify and detect B.animalis.Based on the genomic DNA of B.animalis HP-B1124,the ERIC-PCR reaction conditions of B.animalis HP-B1124 were optimized,and the ERIC-PCR fragments were obtained one by one and sequenced.Two pairs of specific primer probes were designed.The accuracy,specificity,limitation and universality of the two pairs of primer probes were evaluated,and the two pairs of specific primer probes were used for testing the products containing B.animalis in the commercially published formula.The two pairs of specific primer probes designed in this study could be used for identified strains of B.animalis more simply,quickly and targeted.This method has optimized the current relatively traditional methods of pure culture and plate counting identification of B.animalis,and has solved the high requirements of SNP genotyping technology and real-time fluorescence quantitative PCR method for experimental equipment and reagents in the identification of B.animalis to a certain extent.It has the characteristics of low cost,high specificity and earn a broad market development prospect.

关 键 词：动物双歧杆菌 ERIC-PCR 细菌鉴定 引物探针 

分 类 号：R917[医药卫生—药物分析学]