Akhtar N, Eley A
Department of Experimental and Clinical Microbiology, University of Sheffield Medical School, United Kingdom.
J Clin Microbiol. 1992 Sep;30(9):2408-14. doi: 10.1128/jcm.30.9.2408-2414.1992.
Thirty-three clinical isolates from male nongonococcal urethritis and 28 isolates from soft tissue infections and ulcers were identified as Bacteroides ureolyticus by conventional bacteriological tests and were compared with five reference strains of the species. Whole-cell proteins from these clinical isolates and the reference strains were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The majority of the strains from the two sources could be divided into five different groups, named phenons I to V; phenons I to IV have been described previously by others, while phenon V has been described recently by us. Digestion of chromosomal DNA from 16 of the clinical isolates (including strains representative of each of the five SDS-PAGE phenons) and the five reference strains was attempted with restriction endonucleases EcoRI, PstI, SmaI, and HindIII. After electrophoresis in agarose gels, good digestion was observed with HindIII only, and 12 different banding patterns (restriction endonuclease analysis [REA] profiles) were obtained for the 19 strains digested; one nongonococcal urethritis isolate and one reference strain did not show any digestion. From the agarose gels, HindIII-digested fragments of DNA were transferred to nylon membranes by use of vacuum blotting and subjected to hybridization with 32P-labelled 16S-23S rRNA from Escherichia coli. The resultant pattern of bands (ribotypes), which depends on the restriction fragment length polymorphisms in the rRNA genes, was used as a measure of genomic variation within the species. In total, 13 different ribotypes were obtained for the 19 strains. For some strains, good correlation was achieved among the SDS-PAGE phenons, REA profiles, and ribotypes. However, for others, REA analysis and ribotyping were able to discriminate between strains which shared the same SDS-PAGE phenon. Interestingly, these two techniques of DNA characterization were able to differentiate between isolates from the genital tract and those associated with soft tissue infections and ulcers.
通过传统细菌学检测，从男性非淋菌性尿道炎中分离出33株临床菌株，从软组织感染和溃疡中分离出28株菌株，鉴定为解脲拟杆菌，并与该菌种的5株参考菌株进行比较。这些临床分离株和参考菌株的全细胞蛋白通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行分离。来自这两个来源的大多数菌株可分为五个不同的组，命名为表型I至V；表型I至IV先前已被其他人描述过，而表型V最近由我们描述。尝试用限制性内切酶EcoRI、PstI、SmaI和HindIII对16株临床分离株（包括代表五个SDS-PAGE表型的菌株）和5株参考菌株的染色体DNA进行消化。在琼脂糖凝胶中电泳后，仅观察到HindIII有良好的消化效果，对19株消化的菌株获得了12种不同的条带模式（限制性内切酶分析[REA]图谱）；一株非淋菌性尿道炎分离株和一株参考菌株未显示任何消化。从琼脂糖凝胶中，通过真空印迹将HindIII消化的DNA片段转移到尼龙膜上，并用来自大肠杆菌的32P标记的16S-23S rRNA进行杂交。所得的条带模式（核糖型）取决于rRNA基因中的限制性片段长度多态性，用作该菌种内基因组变异的度量。总共为19株菌株获得了13种不同的核糖型。对于一些菌株，SDS-PAGE表型、REA图谱和核糖型之间具有良好的相关性。然而，对于其他菌株，REA分析和核糖分型能够区分具有相同SDS-PAGE表型的菌株。有趣的是，这两种DNA表征技术能够区分生殖道分离株与软组织感染和溃疡相关的分离株。
