特异性检测鼠李糖乳杆菌的引物探针及应用  

Primer probes for specific detection of Lactobacillus rhamnosus and its application

作　　者：李典 杜丽霞 陈文骋 侯亚茹 师文君 李岚芳 侯少阳 LI Dian;DU Li-xia;CHEN Wen-cheng;HOU Ya-ru;SHI Wen-jun;LI Lan-fang;HOU Shao-yang(School of Pharmacy,Heze University,Heze 274000,China)

机构地区：[1]菏泽学院药学院,山东菏泽274000

出　　处：《生物技术》2023年第4期414-421,共8页Biotechnology

基　　金：山东省自然科学基金项目(ZR2022QB146);菏泽学院博士基金(XY21BS35);横向课题(XY22HX02)。

摘　　要：[目的]基于ERIC-PCR技术设计引物探针,用于鼠李糖乳杆菌的特异性检测,探究引物探针的属性特征。[方法]以鼠李糖乳杆菌HP-B1083为研究对象,利用新型ERIC兼并引物与4因素3水平正交实验获得鼠李糖乳杆菌ERIC指纹图谱,根据鼠李糖乳杆菌ERIC序列设计特异性引物探针,并进行引物探针特异性、普适性及检测限度验证。[结果]使用ERIC-PCR技术获得的鼠李糖乳杆菌ERIC指纹图谱,成功设计了4对针对鼠李糖乳杆菌的引物探针。探针能在8种混合益生菌中特异性检出鼠李糖乳杆菌,能高效检出6株鼠李糖乳杆菌的同源菌株,在DNA为27 ng/mL的混合基因组中亦可实现鼠李糖乳杆菌的痕量检出。[结论]设计的4对引物探针能实现鼠李糖乳杆菌的特异性检出,具有高特异性、宽广谱性、强灵敏性的特征,实现DNA为27 ng/mL的混合基因组中鼠李糖乳杆菌的痕量检出,具有良好的探针属性特性。[Objective]The primer probe was designed based on ERIC-PCR technology for the specific detection of Lactobacillus rhamnosus,and the attribute characteristics of the primer probe were explored.[Method]Taking Lactobacillus rhamnosus HP-B1083 as the research object,the ERIC fingerprint of Lactobacillus rhamnosus was obtained by using the new ERIC degenerate primers and four-factor three-level orthogonal experiment.Specific primer probes were designed according to the ERIC sequence of Lactobacillus rhamnosus,and the specificity,universality and detection limit of primer probes were verified.[Result]Four pairs of primer probes for Lactobacillus rhamnosus were successfully designed by using ERIC-PCR technology to obtain the ERIC fingerprint of Lactobacillus rhamnosus.The designed four pairs of primers and probes can specifically detect Lactobacillus rhamnosus in eight mixed probiotics,and can efficiently detect six different strains of Lactobacillus rhamnosus,and trace detection of Lactobacillus rhamnosus can also be achieved in a mixed genome with DNA of 27 ng/mL.[Conclusion]The designed four pairs of primer probes can realize the specific detection of Lactobacillus rhamnosus,which has the characteristics of high specificity,broad spectrum and strong sensitivity.It can realize the trace detection of Lactobacillus rhamnosus in the mixed genome with DNA of 27 ng/mL,and has good probe property characteristics.

关 键 词：ERIC-PCR 鼠李糖乳杆菌 引物探针 特异性 琼脂糖凝胶电泳 4因素3水平正交实验 

分 类 号：Q93-331[生物学—微生物学]