荧光原位杂交法检测双歧杆菌  

FLUORESCENCE IN SITU PROBING OF BIFIDOBACTERIA

作　　者：修淑丽[1] 熊德鑫[1] 杨义[1] 陆莲蓉 

机构地区：[1]解放军304医院创伤中心研究室,北京100037

出　　处：《中国微生态学杂志》2000年第1期3-5,共3页Chinese Journal of Microecology

摘　　要：目的:检验荧光原位杂交法在双歧杆菌属鉴定方面的用途。方法:采用对数生长期的8株双歧杆菌和10株其他厌氧、需氧杆菌在相同的条件下分别与双歧杆菌属特异性16SrRNA寡核苷酸基因探针和细菌界通用16SrRNA寡核苷酸基因探针在载玻片上进行原位杂交,在荧光显微镜下观察杂交结果,拍摄同一视野的荧光显微镜照片和相差显微镜照片,计算杂交率。结果:所用的双歧杆菌菌株均与两种基因探针杂交,在荧光显微镜下杂交菌与黑暗背景对照显著,杂交率在80%以上而其他菌株仅能与细菌界通用探针杂交。结论:所选用的以16SrRNA为靶基因的寡核苷酸探针可用于原位杂交法对双歧杆菌属的鉴定。Objective:To determine the value of fluorescence in situ hybridization (FISH) in identiying genus Bifidobacterium. Methods:Eight strains of Bifidobacterium as well as ten aerobic and anaerobic bacillus in logarrithmic growth phase were used.Hybridization was performed with Bifidobacterium genus-specific 16S rRNA oligonucleotide probe and universal kindom probe under same condition.Hybridization results were observed with OLYMPUS VANOX microscope.The fluorescence photograph and phase-contrast photograph of the same field were taken and hybridizaton percentage was calculated. Results: All Bifidobacteria hybridized with the two probes,their fluorescence contrasted obviousely agaist the background and hybridization percentage was more than80%. Whereas other strains only hybridized with universal-kindom probe. Conclusions: The selected oligonucleotide probe targeting 16S rRNA can be used to identify Bifidobacterium genus using in-situ hybridization methord.

关 键 词：双歧杆菌 检测 荧光原位杂交 16SrRNA 

分 类 号：Q939.139[生物学—微生物学]