Hilton A C, Banks J G, Penn C W
School of Biological Sciences, University of Birmingham, UK.
J Appl Bacteriol. 1996 Dec;81(6):575-84. doi: 10.1111/j.1365-2672.1996.tb03550.x.
Random amplification of polymorphic DNA (RAPD) was evaluated for its ability to differentiate Salmonella strains from various sources. Under defined conditions RAPD using a 10-mer primer (1254) produced a series of amplification products able to reproducibly distinguish strains representing 20 different serotypes of Salmonella. Primer 1254 also proved capable of discrimination between some but not all isolates of Salm. ser. Enteritidis and Salm. ser. Typhimurium, phage typing proving to be most discriminatory for the latter serotype. Cloning of fragments into a vector allowed sequencing and database searching for identification of fragments and an indication of criteria for primer template interaction in RAPD. Southern blotting using a digoxigenin-labelled probe allowed identification of related bands between RAPD profiles. These observations demonstrate the potential of rapid molecular typing by RAPD for the genomic typing of Salmonella strains.
对随机扩增多态性DNA(RAPD)区分不同来源沙门氏菌菌株的能力进行了评估。在特定条件下,使用10聚体引物(1254)的RAPD产生了一系列扩增产物,能够可重复地区分代表20种不同血清型沙门氏菌的菌株。引物1254也被证明能够区分一些但不是所有肠炎沙门氏菌和鼠伤寒沙门氏菌的分离株,噬菌体分型对后一种血清型的区分最为明显。将片段克隆到载体中可进行测序和数据库搜索,以鉴定片段并指示RAPD中引物-模板相互作用的标准。使用地高辛标记探针的Southern印迹法可鉴定RAPD图谱之间的相关条带。这些观察结果证明了通过RAPD进行快速分子分型用于沙门氏菌菌株基因分型的潜力。