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目的比较NotI和SfiI酶切霍乱弧菌基因组DNA进行PFGE指纹图谱分析菌株间遗传关系效果的优劣。方法分别运用NotI和SfiI对2008年在海南省暴发的2起霍乱疫情中分离到的76株菌株进行PFGE指纹分析,并利用BioNumerics Version4.0软件聚类分析。结果 NotI酶切图谱将76株菌株划分为3种不同的带型,SfiI酶切图谱将76株菌株划分为6种不同的带型。两种酶分析的结果均显示,6月份和10～11月份暴发的两起疫情的菌株分别具有不同的带型。SfiI酶切图谱分析显示,6月份疫情中,1株分离自永兴镇的菌株与4株长流镇的菌株间相似系数为98%;10～11月份疫情中3株分离自海南大学的菌株具有不同的带型分别独立成为一支,而NotI酶切图谱中,这些菌株均与同一起疫情中的其他菌株具有相同的带型,未能检测到遗传变异。结论 NotI更适合于对不同暴发事件整体性分析,而SfiI适合于分析暴发事件中菌株间的差异,这两种酶的共同使用起到较好的互补作用,为进一步的防控提供有力的依据。
Objective To compare the genomic DNA of Notch and Vibrio cholerae Vibrio cholerae by PFGE fingerprinting to analyze the genetic relationship between strains. Methods 76 strains isolated from two cholera outbreaks in Hainan Province in 2008 were analyzed by PFGE fingerprinting using NotI and SfiI, respectively. The data were analyzed by BioNumerics Version 4.0 software. RESULTS: Sixty-six isolates were divided into three different bands by NotI digestion, and 76 strains were divided into six different bands by SfiI digestion. The results of the two enzyme assays showed that the strains of the two outbreaks that broke out in June and from October to November each had a different band pattern. SfiI digestion mapping analysis showed that in June the outbreak, a strain isolated from Yongxing town and 4 strains of Changli Town, the similarity coefficient was 98%; 10 to 11 outbreaks in the strains isolated from Hainan University Which belonged to different bands. However, in NotI digestion maps, all the isolates belonged to the same type with other strains in the same epidemic, and no genetic variation was detected. Conclusion NotI is more suitable for the overall analysis of different outbreaks, while SfiI is suitable for analyzing the differences among isolates in outbreaks. The co-use of these two enzymes plays a complementary role and provides a strong basis for further prevention and control .