Development of multi-color FISH method for analysis of seven Bifidobacterium species in human feces
Development of multi-color FISH method for analysis of seven Bifidobacterium species in human feces
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DOI：
10.1016/j.mimet.2004.05.006
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发表时间：
2004-09-01
影响因子：
2.2
通讯作者：
Nomoto, K
中科院分区：
文献类型：
--
作者：
Takada, T;Matsumoto, K;Nomoto, K
我们建立了一种多色荧光原位杂交(FISH)方法，通过一次反应检测所有七种双歧杆菌(双歧杆菌、短链双歧杆菌、短链双歧杆菌、齿双歧杆菌和长双歧杆菌)，它们是人类粪便中的优势细菌。首先，设计了8种新型寡核苷酸探针，与双歧杆菌属和上述每种双歧杆菌特有的16S rRNA序列互补。利用全细胞杂交，针对每个探针所针对的细菌物种测量了荧光强度，表明每个探针针对目标细菌，相对荧光强度(RFI)作为探针可及性的指标很高，为61-117%。然后，用荧光染料(FITC、TAMRA和Cy5)以7种不同的方式单独或组合标记细菌物种特异性探针。使用这些探针，七种双歧杆菌在培养的细菌和成年志愿者粪便的混合样本中进行了差异染色，证明了这项技术的有效性。(C)2004爱思唯尔B.V.保留所有权利。
We have developed a multi-color fluorescence in situ hybridization (FISH) method which detects, by a single reaction, all seven species of Bifidobacterium (B. adolescentis, B. angulation, B. bifidum, B. breve, B. catenulatum, B. dentium, and B. longum), the dominant bacteria in human feces. First, eight new types of oligonucleotide probe were designed, complementary with the 16S rRNA sequence specific to genus Bifidobacterium and each bifidobacterial species described above. Using whole cell hybridization, the fluorescent intensity was measured against the bacterial species targeted by each probe, to show that each probe is specific to the targeted bacteria and that the relative fluorescent intensity (RFI) as an indicator of probe accessibility is high at 61-117%. Then, bacterial species-specific probes were labeled with fluorochromes (FITC, TAMRA and Cy5) in seven different ways, singly or in combination. Using these probes, seven species of Bifidobacterium were differentially stained in mixed samples of cultured bacteria and feces from adult volunteers, proving the efficacy of this technique. (C) 2004 Elsevier B.V. All rights reserved.