Feiková S, Klement C
Regionálny úrad verejného zdravotníctva, Banská Bystrica, SR.
Klin Mikrobiol Infekc Lek. 2007 Dec;13(6):236-41.
Pulsed-field gel electrophoresis was developed for separating and analyzing of long DNA fragments in alternating electric field. In homogenous electric field, fragments longer than 50 kb run as a broad, unresolved band with high mobility. PFGE separated the DNA by periodicaly changing the direction of electric field. DNA molecules are moving "zig-zag" through the gel and they can be better separated. Fragments of several megabases can be resolved using this method. PFGE can be used for genome mapping of microorganisms as well as higher organisms. In microbiology, PFGE is a standard method for typization of bacteria. Comparison of electrophoresis profiles after digestion od DNA from bacterial isolates with restriction endonuclease is a very useful epidemiologists tool. Genetically identical organisms have the same PFGE profiles, different strains have different profiles. Related strains have also similar electrophoretic profiles. This enables to determine if the outbreaks are caused by the same strain of microorganism, to locate the source of outbreak and to monitor the spread of the microorganism. The most followed-up are nosocomial and the food-borne pathogens. PFGE can be also used for monitoring genetic evolution of the microorganism and the most prevalent types which circulate in population. This can be very useful for preparation of vaccines.
脉冲场凝胶电泳是为在交变电场中分离和分析长DNA片段而开发的。在均匀电场中,长度超过50 kb的片段会以高迁移率形成一条宽的、未分离的条带。PFGE通过周期性改变电场方向来分离DNA。DNA分子在凝胶中呈“之字形”移动,从而能得到更好的分离。使用这种方法可以分辨出几兆碱基大小的片段。PFGE可用于微生物以及高等生物的基因组图谱绘制。在微生物学中,PFGE是细菌分型的标准方法。用限制性内切酶消化细菌分离株的DNA后比较电泳图谱是流行病学家非常有用的工具。基因相同的生物体具有相同的PFGE图谱,不同菌株具有不同的图谱。相关菌株也有相似的电泳图谱。这使得能够确定疫情是否由同一菌株的微生物引起,找到疫情源头并监测微生物的传播。最受关注的是医院感染病原体和食源性病原体。PFGE还可用于监测微生物的遗传进化以及在人群中传播的最普遍类型。这对于疫苗制备可能非常有用。