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目的建立与验证创伤弧菌脉冲场凝胶电泳(PFGE)分子分型方法。方法以分子分型监测网络脉冲网(Pulse Net)中公布的霍乱弧菌的PFGE标准方法为基础,选用限制性内切酶NotⅠ和SfiⅠ对36株不同来源的创伤弧菌分离株进行PFGE分型。结果 36株创伤弧菌菌株重复经2种酶切电泳后得到的条带均在25条~30条。有3株菌PFGE电泳条带出现降解,其余33株菌经NotⅠ酶切后,形成32种PFGE型别,图谱相似性为45%~100%,分辨力D值为0.998。33株菌经SfiⅠ酶切后可被完全分型,图谱相似性为50.9%~96.3%,D值为1。结论 PFGE作为一种分辨力高、重复性好的分子分型方法,对创伤弧菌进行PFGE分子分型,优先采用SfiⅠ酶切方法,NotⅠ可以为备选酶,同时结合分离菌株的相关流行病学信息,可作为研究创伤弧菌感染与传播的有力工具。
Objective To establish and validate the molecular typing method of Vibrio vulnificus pulsed-field gel electrophoresis (PFGE). Methods Based on the PFGE standard method of molecular typing monitoring of Vibrio cholerae published in Pulse Net, 36 strains of Vibrio vulnificus isolated from different origins were genotyped by PFGE using restriction endonucleases NotⅠand SfiⅠ . Results 36 strains of Vibrio vulnificus were obtained from 25 to 30 bands after 2 kinds of enzyme digestion electrophoresis. Three strains of PFGE electrophoresis bands were degraded. The remaining 33 strains were digested with NotⅠ to form 32 types of PFGE. The similarity of the bands was 45% -100% and the discrimination value was 0.998. After digestion can be completely sub-type map similarity of 50.9% to 96.3%, D value of 1. Conclusion As a molecular typing method with high resolution and good repeatability, PFGE molecular typing of Vibrio vulnificus was preferentially performed by PFGE, and SfiⅠ digestion was the first choice. NotⅠcan be a candidate enzyme, Learn information, can be used as a powerful tool to study Vibrio vulnificus infection and transmission.