双歧杆菌地高辛标记寡核苷酸基因探针制备和应用研究  

Application and preparation of Bifidobacteria 16S rRNA oligonucleotide probe

作　　者：姚玉川[1] 冯天杰[1] 赵江华[1] 王磊辉 王继德[2] 熊德鑫[3] 

机构地区：[1]解放军152医院消化内科,河南平顶山467000 [2]南方医院消化研究所,广东广州510515 [3]解放军304医院中心研究室,北京100037

出　　处：《中国微生态学杂志》2006年第3期187-188,共2页Chinese Journal of Microecology

摘　　要：目的探讨地高辛标记寡核苷酸基因探针应用于微生态研究的可行性和实用性。方法制备双歧杆菌属和部分种的地高辛标记16S rRNA寡核苷酸探针,初步应用于微生态制剂鉴定和临床肠道微生态检测,评价寡核苷酸探针杂交在肠道微生态研究和检测中的应用价值。结果地高辛标记寡核苷酸探针具有较好的特异性与灵敏度:地高辛标记的双歧杆菌属和种的共6种寡核苷酸基因探针与标准菌株杂交后灵敏度和特异度分别为属探针95%、75%,青春双歧87.5%、90%,两歧双歧87.5%、87.5%,短双歧87.5%、92.5%,婴儿双歧75%、95%,长双歧75%、100%。结论寡核苷酸基因探针用于肠道细菌的鉴定显示出一定前景,加大探针的种类与扩大调查范围有可能使该技术替代现有细菌培养技术。Objective To investgate the application and preparation of Bifidobacteria 16S rRNA oligonucleotide probe. Methods To try to simplify the detection of the imblance of the intestinal microflora, digoxigenin-labeled 16S rRNA oligonucleotide probes of Bifidobacterium genus bacteria were designed and evaluated their sensitivity and specificity in the detection of Bifidobacterium in probiotic preparatios and feces. Results Dig-labeled 16S rRNA oligonucleotide probes of Bifidobacterium had quite high sensitivity and specificity when used to detect Bifidobacterium in probiotics and feces （ strain,95% ,75% ; adolescentis, 87.5% ,90% ; infantis ,75% ,95% ; longum, 7%, 100% ; breve ,87.5% ,92. 5% ; b/fidum 87.5% ,87.5% ）. Conclusion Molecular assays by analyzing 16S rRNA gene of intestinal bacteria should be the promised assay in the evaluation of intestinal microflora dysfunction to replace the classical bacterial culture techniques.

关 键 词：双歧杆菌 菌落杂交 基因探针 16S RRNA 

分 类 号：R378.992[医药卫生—病原生物学]