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目的建立创伤弧菌基因分型的新方法,了解创伤弧菌的分子特征。方法应用PCR-变形梯度凝胶电泳(PCR-DGGE)技术对创伤弧菌的16S-23S rDNA间区(16S-23S rDNA intergenic spacer regions,ISR)多态性进行分析比较,结合药敏试验,以进一步验证分离菌株之间的亲缘关系。结果 ISR-PCR将创伤弧菌菌株扩增出4条约900、750、650、550 bp大小不同的条带;同时,创伤弧菌的ISR-DGGE序列表现出明显的株间差异,18株共产生了16种不同的指纹图谱;聚类分析将所有的细菌分为两大类,相似性分别为0.65和0.71;亲缘关系较近的菌株具有不同于关系较远的菌株的耐药谱,与ISR-DGGE指纹图谱分析相一致。结论这种分子操作技术完全能够用于创伤弧菌株型的调查与鉴定,为创伤弧菌的基因分型提供了一种新的方法。
Objective To establish a new method of genotyping Vibrio vulnificus and understand the molecular characteristics of Vibrio vulnificus. Methods The 16S-23S rDNA intergenic spacer regions (ISR) polymorphisms of Vibrio vulnificus were analyzed by PCR-DGGE and PCR-DGGE. Further verify the genetic relationship between isolates. Results ISR-PCR amplified four Vibrio vulnificus strains with different sizes of 900, 750, 650 and 550 bp. At the same time, ISR-DGGE sequences of Vibrio vulnificus showed obvious inter-strain differences and 18 strains were co-produced 16 different fingerprints were obtained. Cluster analysis divided all the bacteria into two groups with similarities of 0.65 and 0.71, respectively. The more closely related strains had different resistance spectrum than the strains with ISR -DGEGE fingerprinting analysis. Conclusion This molecular manipulation technique can be completely used in the investigation and identification of Vibrio vulnificus. It provides a new method for genotyping Vibrio vulnificus.