NotI和SfiI酶切PFGE指纹分析比较  

Comparison of PFGE fingerprinting between NotI enzyme and SfiI

作　　者：吴捷[1] 朱建华[1] 刁保卫[2] 周海建 阚飙[2] 黄昌和[1] 王少玲[1] 苏新元[1] 马焱[1] 

机构地区：[1]海南省疾病预防控制中心,海南海口570203 [2]中国疾病预防控制中心传染病预防控制所中国疾病预防控制中心传染病预防控制国家重点实验室

出　　处：《现代预防医学》2012年第12期3054-3057,3061,共5页Modern Preventive Medicine

基　　金：海南省卫生厅科学研究课题(琼卫2009-2)

摘　　要：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 use NotI enzyme digestion and SfiI enzyme digestion of the genome DNA of Vibrio Cholera to conduct the PFGE fingerprinting analysis about genetic relation among strains, and compare the effect of the two enzyme. METHODS NotI and SfiI were respectively used for PFGE fingerprinting analysis of 76 strains which were isolated from 2 outbreaks of cholera in 2008, and analyze the data with BioNumerics Version 4.0. RESULTS Nod Macrorestriction Map divided those 76 strains to 3 banding patterns, while SfiI＇s were 6 banding patterns. Both results showed that there were different banding pat- terns between strains isolated from 2 outbreaks. One outbreak occurred in June and another one was during October to November. According to the results of Sill Maerorestrietion Map, and the similarity factor was 98%, and one strain was isolated from Yong Xing Town and anther four strains were from Chang Lin Town in June; There were different banding patterns among 3 strains isolated from Hainan University in the outbreak during October to November. However, the results of Notl Macrorestriction Map showed that the banding patter of those strains mentioned above was the same as the others from one outbreak, and there was no genetic variation. CONCLUSION Not/is better for the integrity analysis about different outbreak events, while SfiI is fit for the analysis about the diversity among strains of outbreak. The complementation of the two enzymes provides the scientific support for further prevention and control of infection.

关 键 词：霍乱弧菌 脉冲场凝胶电泳 NotI酶切 SfiI酶切 

分 类 号：R378.3[医药卫生—病原生物学]