Department of Epidemiology and Clinical Microbiology, National Medicines Institute, Warszawa, Poland.
Virulence. 2012 Oct 1;3(6):534-8. doi: 10.4161/viru.21887.
We recently developed a method that allows fast differentiation between Streptococcus pyogenes (GAS) strains. The method named phage profiling (PP) is based on a simple assumption that a regular PCR reaction with Taq polymerase and relatively short elongation time is not able to yield long DNA fragment, such as ~40-50 kb integrated prophage. Only fragments without any integrated DNA or short fragments inserted between integration sites can be efficiently amplified. We designed primers that anneal upstream and downstream prophage integration sites, so in simple PCR reaction we can test if any additional DNA is integrated into particular site. Profiling of integrated elements can be used as rapid, high resolution typing method, with the resolution as high as PFGE and is excellent predictor of PFGE type.
我们最近开发了一种方法,可以快速区分酿脓链球菌(GAS)菌株。该方法名为噬菌体分析(PP),基于一个简单的假设,即使用 Taq 聚合酶的常规 PCR 反应和相对较短的延伸时间不能产生长的 DNA 片段,例如~40-50 kb 整合的前噬菌体。只有没有任何整合 DNA 的片段或插入整合位点之间的短片段才能被有效扩增。我们设计了与前噬菌体整合位点上下游退火的引物,因此在简单的 PCR 反应中,我们可以测试任何额外的 DNA 是否整合到特定的位点。整合元件的分析可以作为一种快速、高分辨率的分型方法,分辨率高达 PFGE,并且是 PFGE 类型的优秀预测因子。