PDF(Pubmed)
Abstract：
For decades now, DNA fingerprinting by means of pulsed-field gel electrophoresis (PFGE) continues to be the most widely used to separate large DNA molecules and distinguish between different strains in alternating pulses. This is done by isolating intact chromosomal DNA and using restriction enzymes with specific restriction sites to generate less than 30 restriction fragments from 50 Kb to 10 Mbp. These results make clone-specific band profiles easy to compare. Specialized equipment is required for the optimization of DNA separation and resolution, among which a contour-clamped homogeneous electric field (CHEF) apparatus is the most commonly used. As a result, the PFGE analysis of a bacterial genome provides useful information in terms of epidemiological investigations of different bacterial pathogens. For Staphylococcus aureus subtyping, despite its limitations and the emergence of alternative methods, PFGE analysis has proven to be an adequate choice and the gold standard for determining genetic relatedness, especially in outbreak detection and short-term surveillance in the veterinary field.
摘要：
几十年来，通过脉冲场凝胶电泳（PFGE）进行的DNA指纹分析仍然是最广泛用于分离大DNA分子并区分交替脉冲中的不同菌株的方法。这是通过分离完整的染色体DNA并使用具有特定限制性位点的限制性酶以产生少于30个50Kb至10Mbp的限制性片段来完成的。这些结果使得克隆特异性条带谱易于比较。需要专门的设备来优化DNA分离和分辨率，其中轮廓钳制均匀电场（CHEF）设备是最常用的。因此,对细菌基因组的PFGE分析为不同细菌病原体的流行病学调查提供了有用的信息.对于金黄色葡萄球菌亚型，尽管它的局限性和替代方法的出现，PFGE分析已被证明是确定遗传相关性的适当选择和黄金标准，特别是在兽医领域的疫情检测和短期监测方面。
