环介导等温扩增技术快速检测双歧杆菌属细菌

Rapid detection of Bifidobacterium spp. by loop-mediated isothermal amplification

中文关键词:  双歧杆菌  环介导等温扩增  特异性  16S rDNA基因

英文关键词:Bifidobacterium  loop-mediated isothermal amplification  specificity  16S rDNA gene

中文摘要:

      目的 建立一种利用环介导等温扩增(LAMP)技术快速检测双歧杆菌属细菌的方法。方法 针对双歧杆菌属细菌16S rDNA基因特异性区段设计LAMP引物, 根据反应体系的颜色或浊度变化特异性地检测样品中双歧杆菌属细菌。结果 对5株双岐杆菌属细菌和19株非双歧杆菌属细菌进行特异性和非特异检测, 结果均为100%。该检测可在60 min内完成, 双歧杆菌属细菌DNA的检测限为16.1±8.2 pg/μL, 对双歧杆菌属细菌菌体的检测限为0.1 CFU/mL。结论 LAMP方法可作为传统国家标准检测方法的辅助手段, 有效检测食品、保健食品和药品中双歧杆菌属细菌DNA的存在。

英文摘要:

      Objective To develop a specific and rapid method for detection of Bifidobacterium spp. by loop-mediated isothermal amplification (LAMP). Methods Specific DNA fraction of 16S rDNA gene of Bifidobacterium spp. was targeted for primer design of LAMP. Bifidobacterium spp. was confirmed by the changes of color or turbidity of reactions specifically. Results Five strains of Bifidobacterium spp. and 19 strains of non-Bifidobacterium spp. were tested respectively in 60 min. The specificity and exclusivity of the method were 100%. The detection limit of DNA from Bifidobacterium spp. was 16.1±8.2 pg/μL, and the detection limit of thallus from Bifidobacterium spp. was 0.1 CFU/mL. Conclusion As a complementary method of national standard, the LAMP can be used to test Bifidobacterium spp. in foods, health foods and pharmaceutical products.