Stefańska Ilona, Kwiecień Ewelina, Górzyńska Małgorzata, Sałamaszyńska-Guz Agnieszka, Rzewuska Magdalena
Department of Preclinical Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Ciszewskiego 8 St., 02-786 Warsaw, Poland.
Pathogens. 2022 May 10;11(5):562. doi: 10.3390/pathogens11050562.
In this study, a Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) method for genetic typing of , an opportunistic bacterial pathogen, was designed. The method optimization was performed for 37 clinical strains isolated from various infections in different animal species. Optimal conditions for reliable and reproducible DNA fingerprinting were determined according to the modified Taguchi method. The developed method was assessed regarding its typeability, reproducibility, and discriminatory power using the Hunter's and Gatsons' index of discrimination. A high degree of genetic diversity was shown between the studied strains, which represented 31 genotypes. The generated RAPD profiles were relatively complex and simultaneously easy to interpret due to the wide size range of amplicons. The discriminatory index of the designed method was sufficiently high; thus, only strains epidemiologically related displayed identical RAPD genotypes. In conclusion, the DNA fingerprinting of by the developed RAPD-PCR method is a reliable typing tool that may allow a better understanding of the epidemiology as well as pathogenesis of infections caused by this pathogen.
在本研究中,设计了一种用于对机会性细菌病原体进行基因分型的随机扩增多态性DNA - 聚合酶链反应(RAPD - PCR)方法。对从不同动物物种的各种感染中分离出的37株临床菌株进行了方法优化。根据改进的田口方法确定了可靠且可重复的DNA指纹图谱的最佳条件。使用亨特和加特森鉴别指数评估所开发方法的分型能力、重现性和鉴别力。研究菌株之间显示出高度的遗传多样性,代表了31种基因型。由于扩增子的大小范围广泛,所生成的RAPD图谱相对复杂且同时易于解读。所设计方法的鉴别指数足够高;因此,只有在流行病学上相关的菌株显示出相同的RAPD基因型。总之,通过所开发的RAPD - PCR方法对该病原体进行DNA指纹图谱分析是一种可靠的分型工具,可能有助于更好地理解由该病原体引起的感染的流行病学以及发病机制。