Development of monitaruin system for intestinal flora of experimental animals using FISH method

根据分离的梭菌16 SrDNA序列设计了4个组特异性探针，并从小鼠粪便中直接克隆了DNA。为了检测乳杆菌，制备了12种探针，Lab 722（乳杆菌组）、Labc（嗜酸乳杆菌）、Lbam（嗜淀粉乳杆菌）、Lber（卷曲乳杆菌）、Lbga（加氏乳杆菌）、Lbin（嗜酸乳杆菌）、Lbjg（约氏乳杆菌/加氏乳杆菌）、Lbjh（约氏乳杆菌）、Lbmu（鼠乳杆菌/动物乳杆菌）、Lbpa（副干酪乳杆菌）、Lbre（罗伊氏乳杆菌）、Lbva（迷走乳杆菌）。本研究利用16 S和23 S rDNA序列设计了Lbin、Lbjg、Lbmu、Lbva四个基因。对于拟杆菌属，9个探针用于拟杆菌属物种，2个探针用于2个拟杆菌属组。拟杆菌群的Bac 303、拟杆菌群的Bfa 602、酸粪B的Baf 1001、狄氏B的Bdis 656、脆弱B的Bfra 998、卵形B的Bova、多形B的Bth 172、单形B的Buni、普通B的Bvul。本研究设计了Baf 1001和Bth 172，前期设计了Bova、Buni和Buni。这些探针显示出高特异性，并使用FISH检测小鼠粪便中的各组。

We designed four group specific probes for Clostridium based on the 16S rDNA sequenc3es of isolates and directly cloned DNA from mouse feces. For detection of Lactobacillus 12 probes were prepared, Lab722 (Lactobacillus group), Labc (L.acidophilus), Lbam (L.amylovorus), Lber (L.crispatus), Lbga (L.gasseri), Lbin (L.intestinalis), Lbjg (L.johnsonii/L.gasseri), Lbjh (L.johnsonii), Lbmu (L.murinus/L.animalis), Lbpa (L.paracasei), Lbre (L.reuteri), Lbva (L.vaginalis). Lbin, Lbjg, Lbmu, Lbva were designed in this study using 16S and 23S rDNA sequences. For Bacteroides, nine probes for Bacteroides species and two probes for two Bacteroides groups. They were Bac303 for Bacteroides cluster, Bfa602 for Bacteroides group, Baf1001 for B.acidofaeciens, Bdis656 for B.distasonis, Bfra998 for B.fragilis, Bova for B.ovatus, Bth172 for B.thetaiotaomicron, Buni for B.uniformis, Bvul for B.vulgatus. Baf1001 and Bth172 were designed in this study and Bova, Buni and Buni were designed in our previous study. These probes showed high specificity and detected each group in mouse feces using FISH.