嗜水气单胞菌分离株ERIC-PCR及RAPD分型比较

Typing of Aeromonas hydrophila isolates by ERIC-PCR and RAPD

采用ERIC-PCR和RAPD 2种方法对嗜水气单胞菌2009年分离菌株及实验室保存菌株共83株进行分子分型。参考相关文献设计并合成引物,分别进行PCR扩增及琼脂糖凝胶电泳,电泳结果经Quantity One软件数值化后利用SPSS软件进行聚类分析。结果显示,除少数分离株外,使用2种方法对大多数菌株分型,均能得到多态性较好的指纹图谱。RAPD的AP12H引物能扩增出1～12个200～4 500bp之间的条带,可将嗜水气单胞菌分离株分为4群、12类。ERIC-PCR能扩增出4～16个100～5 800bp之间的条带,将嗜水气单胞菌分离株分为4群、17类,同一年代、同一地域分离株分别有聚成一类的趋势。2种分型方法均体现很好的分型能力。但ERIC-PCR分型与RAPD分型相比,重复性和稳定性更好,分辨力更高,且具备RAPD不要求模板序列已知而直接进行扩增的优点,更适合嗜水气单胞菌的分型研究。

83 Aeromonas hydrophila isolates was typed using ERIC-PCR and RAPD.The results of agarose gel electrophoresis were figured by the Quantity One software,and then clustered by SPSS.The results showed that except few strains,most of the A.hydrophila isolates obtained the well-polymorphism fingerprints using the two methods.One to 12 bands between 200 bp and 4 500 bp were amplified by RAPD using the AP12H primer,and the A.hydrophila strains could be divided into 4 main groups and 12 subdivisions.Four to 16 bands between 100 bp and 5 800 bp were amplified by ERIC-PCR,and the A.hydrophila strains could be divided into 4 main groups and 17 subdivisions.There was a tendency that the strains from the same time or geographic area could be clustered into one group.The two methods showed good typing capability.But ERIC-PCR had more reproducibility and stability than RAPD,whick could amplify template unkown its sequences.Therefore ERIC-PCR was more suitable to type A.hydrophila strains.