Appl Environ Microbiol. 1997 Jul;63(7):2600-6. doi: 10.1128/aem.63.7.2600-2606.1997.
The intraspecific genomic relatedness of 80 Vibrio vulnificus isolates, 44 of biotype 1 and 36 of biotype 2, from different geographic origins and sources was evaluated by ribotyping and AFLP (amplified fragment length polymorphism) fingerprinting. Ribopatterns of DNAs digested with KpnI and hybridized with an oligonucleotide complementary to a highly conserved sequence in the 23S rRNA gene revealed up to 19 ribotypes in the species, which were different for the two biotypes. Sixteen different ribotypes were found within biotype 1 strains from clinical and environmental sources, and only three, recovered mainly from diseased eels, were found within biotype 2. Within this biotype, 96% of the strains showed the same ribopattern. The closest similarity was shown by the strains coming from the same eel farm, irrespectively of biotype. AFLP fingerprints obtained by selective PCR amplification of HindIII-TaqI double-restricted DNA fragments exhibited a strain-specific pattern which allowed the finest differentiation of subgroups within the eel-pathogenic isolates sharing the same ribopattern. Both techniques revealed good genetic markers for intraspecific differentiation of V. vulnificus. Ribotyping clearly separated the eel-pathogenic strains from the clinical and environmental isolates, whereas AFLP enabled the monitoring of individual strains and therefore constitutes one of the most discriminative tools for epidemiological and ecological studies.
通过核糖体分型和 AFLP（扩增片段长度多态性）指纹分析，评估了来自不同地理来源和来源的 80 株创伤弧菌（Vibrio vulnificus）分离株的种内基因组相关性，其中 44 株为生物型 1，36 株为生物型 2。用 KpnI 消化的 DNA 的核糖体图谱，并与 23S rRNA 基因中高度保守序列的寡核苷酸杂交，显示该物种中有多达 19 种核糖体型，两种生物型之间存在差异。在临床和环境来源的 1 型菌株中发现了 16 种不同的核糖体型，而在主要从患病鳗鱼中分离的 2 型菌株中仅发现了 3 种。在该生物型中，96%的菌株显示出相同的核糖体型。来自同一鳗鱼养殖场的菌株显示出最接近的相似性，与生物型无关。通过 HindIII-TaqI 双限制 DNA 片段的选择性 PCR 扩增获得的 AFLP 指纹显示出菌株特异性模式，允许对具有相同核糖体型的鳗鱼致病性分离株内的亚群进行最精细的分化。这两种技术都为创伤弧菌的种内分化提供了良好的遗传标记。核糖体分型清楚地将鳗鱼致病性菌株与临床和环境分离株分离，而 AFLP 能够监测单个菌株，因此是流行病学和生态学研究中最具区分力的工具之一。
