Brachyspira aalborgi infection diagnosed by culture and 16S ribosomal DNA sequencing using human colonic biopsy specimens
Brachyspira aalborgi infection diagnosed by culture and 16S ribosomal DNA sequencing using human colonic biopsy specimens
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DOI：
10.1128/jcm.38.10.3555-3560.2000
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发表时间：
2000-10-01
影响因子：
9.4
通讯作者：
Fellström, C
中科院分区：
文献类型：
--
作者：
Kraaz, W;Pettersson, B;Fellström, C
在这项研究中，我们报告了使用取自患有肠道螺旋体病的年轻成年男性结肠的人类粘膜活检标本对肠道螺旋体阿尔伯格短螺旋体的分离和表征。将含有400 μ g壮观霉素/ml和5 μ g多粘菌素/ml的选择性培养基用于分离过程。在表型性状和16 S核糖体DNA序列方面，观察到分离的菌株W1与B的模式菌株513 A之间具有高度的相似性。阿尔博吉W1与513 A（T）的核苷酸序列同源性为99.7%。16 S rRNA基因的一个短片段通过PCR扩增，使用从石蜡包埋的活检标本，这是从病人的两个场合的遗传物质富集。产物显示16 S rRNA基因序列在实际区域中与菌株513 A（T）的序列几乎相同。免疫组织化学进行结肠活检标本与多克隆抗体提出了对肠道螺旋体分离在以前的情况下，人类肠道螺旋体病。抗体与腔上皮上的螺旋体反应强烈。在表面上皮内或下方未观察到免疫反应。常规组织学检查未发现结肠炎体征。电子显微镜检查显示螺旋体末端附着于结肠粘膜表面。该分离株在肠螺旋体常用的选择性培养基上生长不良，这可能解释了以前未能分离出B，aalborgi的原因。
In this study we report on the isolation and characterization of the intestinal spirochete Brachyspira aalborgi using human mucosal biopsy specimens taken from the colon of a young adult male with intestinal spirochetosis. A selective medium, containing 400 mu g of spectinomycin/ml and 5 mu g of polymyxin/ml was used for the isolation procedure. A high degree of similarity, in terms of phenotypic properties and 16S ribosomal DNA sequence, was observed between the isolated strain, named W1, and the type strain, 513A, of B. aalborgi. A similarity of 99.7% in the nucleotide sequence was found between W1 and 513A(T), based on the almost-complete gene. A short segment of the 16S rRNA gene was amplified by PCR using genetic material enriched from paraffin-embedded biopsy specimens, which were taken from the patient on two occasions. The products showed 16S rRNA gene sequences virtually identical to that of strain 513A(T) in the actual region. Immunohistochemistry was performed on the colonic biopsy specimens with a polyclonal antibody raised against an intestinal spirochete isolated in a previous case of human intestinal spirochetosis. The antibody reacted strongly with the spirochete on the luminal epithelium. No immune reaction was seen within or below the surface epithelium. Routine histology did not reveal signs of colitis. Electron microscopy showed spirochetes attached end-on to the colonic mucosal surface. The isolate grew poorly on a commonly used selective medium for intestinal spirochetes, which may explain previous failures to isolate B, aalborgi.