Fox J G, Dewhirst F E, Fraser G J, Paster B J, Shames B, Murphy J C
Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge 02139.
J Clin Microbiol. 1994 May;32(5):1229-37. doi: 10.1128/jcm.32.5.1229-1237.1994.
Proliferative bowel disease is an intestinal disorder of a variety of domestic animals associated with the presence of an intracellular Campylobacter-like organism (ICLO). We have identified the ICLO obtained from a ferret with proliferative colitis by 16S rRNA sequence analysis. In this ferret, proliferative bowel tissue containing the ICLO had translocated to the mesenteric lymph nodes, omentum, and liver. The 16S rRNA genes of the ICLO were amplified from an infected fragment of extraintestinal tissue by using universal prokaryotic primers. Approximately 1,480 bases of the amplified 16S rRNA gene were sequenced by cycle sequencing. Comparison of the sequence of the ICLO with those of over 400 bacteria in our data base indicated that the sequence of the ICLO was most closely related to that of Desulfovibrio desulfuricans (87.5% similarity). Phylogenetic analysis with 12 Desulfovibrio species and 20 species from related genera placed the ICLO in a subcluster within the genus Desulfovibrio with D. desulfuricans and 5 other Desulfovibrio species. We will refer to this organism as the intracellular Desulfovibrio organism (IDO). Specific primers were produced for PCR amplification of a 550-base fragment of the 16S rRNA gene of the IDO in proliferative intestinal tissue samples. This unique 550-base segment was amplified from samples of frozen intestinal tissue from nine ferrets and three hamsters with ICLO-associated disease but not in four intestinal tissue samples from animals without the ICLO-associated disease. The 550-base amplified products from the bowel tissues of one hamster and one ferret were fully sequenced. The ferret IDO partial sequence was identical to the previously determined 16S rRNA sequence over its length, and the hamster IDO sequence differed by a single base. The same intracellular organism has been identified in proliferative intestinal tissues of swine and that the organism has been successfully maintained in tissue culture. The availability of specific primers for PCR-based detection of this intracellular Desulfovibrio organism will aid in the determination of its role in the pathogenesis of proliferative bowel disease in a variety of infected hosts.
增生性肠病是一种与细胞内弯曲杆菌样生物体(ICLO)存在相关的多种家畜肠道疾病。我们通过16S rRNA序列分析鉴定了从一只患有增生性结肠炎的雪貂身上获得的ICLO。在这只雪貂中,含有ICLO的增生性肠组织已转移至肠系膜淋巴结、网膜和肝脏。通过使用通用原核引物,从肠外组织的感染片段中扩增出ICLO的16S rRNA基因。通过循环测序对扩增的16S rRNA基因的约1480个碱基进行了测序。将ICLO的序列与我们数据库中400多种细菌的序列进行比较,结果表明ICLO的序列与脱硫脱硫弧菌的序列关系最为密切(相似性为87.5%)。对12种脱硫弧菌属物种和来自相关属的20种物种进行系统发育分析,结果将ICLO置于脱硫弧菌属内的一个亚群中,与脱硫脱硫弧菌和其他5种脱硫弧菌属物种在一起。我们将这种生物体称为细胞内脱硫弧菌生物体(IDO)。制备了特异性引物,用于PCR扩增增生性肠组织样本中IDO的16S rRNA基因的550个碱基片段。从9只雪貂和3只患有ICLO相关疾病的仓鼠的冷冻肠组织样本中扩增出了这个独特的550个碱基片段,但在4只没有ICLO相关疾病的动物的肠组织样本中未扩增出。对一只仓鼠和一只雪貂的肠组织中的550个碱基扩增产物进行了全序列测定。雪貂IDO部分序列在其全长上与先前确定的16S rRNA序列相同,仓鼠IDO序列有一个碱基不同。在猪的增生性肠组织中也鉴定出了相同的细胞内生物体,并且该生物体已在组织培养中成功保存。用于基于PCR检测这种细胞内脱硫弧菌生物体的特异性引物的可用性将有助于确定其在多种受感染宿主的增生性肠病发病机制中的作用。