Hernández G, Olmos J
Molecular Microbiology Laboratory, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Department of Marine Biotechnology, Ensenada, B.C. México.
Appl Microbiol Biotechnol. 2004 Feb;63(6):722-7. doi: 10.1007/s00253-003-1451-z. Epub 2003 Oct 21.
Fifteen environmental samples of Vibrio spp. isolated from healthy and diseased shrimps were tested for pathogenicity to juvenile shrimps. Two isolates, strains Z2 and Z3, were observed to be pathogenic, causing 100% mortality of the target host compared to the control strain Vibrio harveyi ATCC 14126. Environmental and type strains were subjected to molecular characterization by restriction fragment length polymorphism (RFLP) and PCR using primers targeted to different virulence, transcriptional regulator, or quorum sensing genes from V. harveyi. Primers designed for luxN were specific and identified all the environmental strains as V. harveyi. The random amplified polymorphic DNA (RAPD) method was used to differentiate between pathogenic and nonpathogenic strains of V. harveyi. These methodologies allowed us to detect and distinguish strains virulent and avirulent to juvenile shrimp.
对从健康和患病虾中分离出的15株弧菌属环境菌株进行了对幼虾的致病性测试。观察到两株菌株Z2和Z3具有致病性，与对照菌株哈维氏弧菌ATCC 14126相比，导致目标宿主100%死亡。通过限制性片段长度多态性（RFLP）和使用针对哈维氏弧菌不同毒力、转录调节因子或群体感应基因的引物进行PCR，对环境菌株和模式菌株进行了分子特征分析。为luxN设计的引物具有特异性，将所有环境菌株鉴定为哈维氏弧菌。采用随机扩增多态性DNA（RAPD）方法区分哈维氏弧菌的致病菌株和非致病菌株。这些方法使我们能够检测和区分对幼虾有毒力和无毒力的菌株。
