He Yiping, Xie Yanping, Reed Sue
Molecular Characterization of Foodborne Pathogens Research Unit, US Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, PA, USA.
Methods Mol Biol. 2014;1085:103-11. doi: 10.1007/978-1-62703-664-1_6.
Pulsed-field gel electrophoresis (PFGE) is the most applied and effective genetic typing method for epidemiological studies and investigation of foodborne outbreaks caused by different pathogens, including Staphylococcus aureus. The technique relies on analysis of large DNA fragments generated by the cleavage of intact bacterial chromosomes with a rare cutting restriction enzyme, subsequently resolved by pulsed-field electrophoresis with periodic changes of the orientation of the electrical field across the gel. The high discriminatory power, improved reproducibility due to standardization of experimental protocols and data interpretation guidelines, and establishment of a national PFGE database of S. aureus profiles have made it a valuable means for global tracking of S. aureus infection sources and determination of genetic relatedness of outbreak isolates.
脉冲场凝胶电泳(PFGE)是流行病学研究以及调查包括金黄色葡萄球菌在内的不同病原体引起的食源性疾病暴发时应用最广泛且有效的基因分型方法。该技术依赖于用一种稀有切割限制酶切割完整细菌染色体产生的大片段DNA的分析,随后通过脉冲场电泳进行分离,电场方向在凝胶上周期性变化。高分辨力、因实验方案和数据解释指南标准化而提高的重现性,以及建立金黄色葡萄球菌谱的国家PFGE数据库,使其成为全球追踪金黄色葡萄球菌感染源和确定暴发分离株遗传相关性的宝贵手段。