椰毒假单胞菌酵米面亚种脉冲场凝胶电泳分型的研究  

Molecular Typing of Pseudomonas cocovenenans subsp.farinofermentans with Pulsed-Field Gel Electrophoresis

作　　者：郭云昌[1] 王岗[1] 杜春明[1] 付萍[1] 裴晓燕[1] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所,北京100021

出　　处：《中国食品卫生杂志》2010年第3期236-239,共4页Chinese Journal of Food Hygiene

基　　金：国家自然科学基金青年基金(30901200)

摘　　要：目的建立椰毒假单胞菌酵米面亚种的脉冲场凝胶电泳(pulsed-field gel electrophoresis,PFGE)分型方法,并对29株分离株进行PFGE分型研究。方法分别选择5种限制性内切酶(XbaⅠ、SpeⅠ、NotⅠ、SmaⅠ、AscⅠ)酶切椰毒假单胞菌酵米面亚种染色体DNA以进行PFGE分析,并利用BioNumerics软件对分离株的指纹图谱进行聚类分析。结果NotⅠ、SmaⅠ和AscⅠ的酶切片段过小,而XbaⅠ酶切片段的大小适中但数量过多,不能获得清晰可辨的图谱,均不适于椰毒假单胞菌酵米面亚种的PFGE分型。SpeⅠ酶切的PFGE条带数量和大小适中、指纹图谱清晰可辨,对椰毒假单胞菌酵米面亚种具有足够的分辨率。结论本研究建立的PFGE分型方法可应用于椰毒假单胞菌酵米面亚种的分子分型和溯源。Objective To establish a method for molecular typing Pseudomonas cocovenenans subsp.farinofermentans with pulsed-field gel electrophoresis（PFGE）,and to analyze the type of 29 isolates with PFGE method.Method The isolates were digested with 5 kinds of restriction endonucleases XbaⅠ,SpeⅠ,NotⅠ,SmaⅠ and AscⅠ,and the PFGE patterns were analyzed by BioNumerics software.Results The bands on PFGE for the fragments digested by restriction endonucleases NotⅠ,SmaⅠ or AscⅠwere too small,and that digested by XbaⅠ were too many to identify clearly.The restriction endonucleases NotⅠ,SmaⅠ,Asc Ⅰ and XbaⅠwere not suitable for typing the pathogen on PFGE.The number of bands on PFGE for the fragments digested by SpeⅠ was moderate and the pattern of molecular weight was clear and well discriminated.Conclusion This method can be used for molecular typing and tracing Pseudomonas cocovenenans subsp.farinofermentans.

关 键 词：椰毒假单胞菌酵米面亚种 脉冲场凝胶电泳 分子分型 

分 类 号：R155.3[医药卫生—营养与食品卫生学]