Terletski Valeri, Schwarz Stefan, Carnwath Joseph, Niemann Heiner
All Russian Research Institute for Animal Genetics & Breeding, 196600 St.Petersburg-Pushkin, Russia.
Microbiol Res. 2003;158(2):135-42. doi: 10.1078/0944-5013-00191.
The aim of the present study was to evaluate the usefulness of a new typing technique called subtracted restriction fingerprinting (SRF) for bacterial strain and isolate discrimination. The technique was applied to isolates of Salmonella enterica subsp. enterica (S.) serovars Choleraesuis, Typhimurium, Dublin and to two laboratory strains of E. coli. SRF is based on the selective removal of excess fragments from a restriction digest using magnetic particles. Subsequently, the remaining subset of restriction fragments can easily be analyzed with a conventional agarose gel. Larger fragments are preferentially removed by SRF. This results in an even distribution of bands within each electrophoretic lane and significantly improves scoring. The high discriminatory index for (S.) Choleraesuis (D = 0.914) illustrated the suitability of SRF for genome typing.
本研究的目的是评估一种名为减法限制性指纹图谱(SRF)的新分型技术在区分细菌菌株和分离株方面的实用性。该技术应用于肠炎沙门氏菌肠炎亚种(S.)霍乱血清型、鼠伤寒血清型、都柏林血清型的分离株以及两株大肠杆菌实验室菌株。SRF基于使用磁性颗粒从限制性酶切消化产物中选择性去除多余片段。随后,剩余的限制性片段子集可以很容易地用传统琼脂糖凝胶进行分析。较大的片段优先被SRF去除。这导致每个电泳泳道内条带分布均匀,并显著提高了评分。霍乱血清型(S.)的高鉴别指数(D = 0.914)表明SRF适用于基因组分型。