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

我们根据分离株的16S rDNA序列设计了4组针对梭状芽胞杆菌的特异性探针，并直接从小鼠粪便中克隆了DNA。制备了Lab722(乳杆菌群)、LABC(嗜酸乳杆菌)、Lbam(淀粉乳杆菌)、Lber(卷曲乳杆菌)、Lbga(乳杆菌)、Lbin(肠杆菌)、Lbjg(约翰逊乳杆菌/加瑟氏乳杆菌)、Lbjh(约翰逊乳杆菌/加瑟氏乳杆菌)、Lbmu(嗜酸乳杆菌/动物乳杆菌)、Lbpa(副乳杆菌)、Lbre(路氏乳杆菌)、Lbva(阴道乳杆菌)等12种乳杆菌探针。本研究利用16S和23S rDNA序列设计了Lbin、Lbjg、LbMu、Lbva。对于类杆菌，9个探针用于类杆菌物种，2个探针用于两个类杆菌类群。它们分别是：Bac303，Bfa602，B.acidofaeciens，Baf1001，B.disasonis，Bdis656，Bfra998，B.ovatus，Bth172，Bth172，B.uniformis，Bvul，Bvul。Baf1001和Bth172在本研究中设计，Bova、Buni和Buni在我们先前的研究中设计。这些探针表现出高度的特异性，并用FISH方法在小鼠粪便中检测到每一组。

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.