Dendani Chadi Zoubida, Arcangioli Marie-Anne
Laboratory of Biodiversity and Pollution of Ecosystems, Department of Veterinary Medicine, Faculty of Natural Science and Life, University of Chadli Bendjedid, El Tarf 36000, Algeria.
VetAgro Sup, Université de Lyon, UMR Mycoplasmoses Animales, 69280 Marcy l'Etoile, France.
Pathogens. 2023 Jul 24;12(7):966. doi: 10.3390/pathogens12070966.
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 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.
几十年来,通过脉冲场凝胶电泳(PFGE)进行DNA指纹识别一直是分离大型DNA分子并在交替脉冲中区分不同菌株的最广泛使用的方法。这是通过分离完整的染色体DNA并使用具有特定限制位点的限制酶来产生50 Kb至10 Mbp的少于30个限制片段来实现的。这些结果使得克隆特异性条带图谱易于比较。优化DNA分离和分辨率需要专门的设备,其中轮廓钳制均匀电场(CHEF)装置是最常用的。因此,细菌基因组的PFGE分析在不同细菌病原体的流行病学调查方面提供了有用的信息。对于亚型分析,尽管存在局限性且出现了替代方法,但PFGE分析已被证明是确定遗传相关性的适当选择和金标准,特别是在兽医领域的疫情检测和短期监测中。