RAPD技术在志贺菌及沙门菌鉴别中的应用

Study on Indentification of Shigella and S.Pullorum by Random Amplified Polymorphic DNA Technology

以鸡鲍氏志贺菌、鸡白痢沙门菌和痢疾志贺菌为研究对象,分别提取基因组DNA,利用6条随机引物以随机扩增多态性DNA(RAPD)技术对其基因组DNA进行了分析.结果表明,4条随机引物能较好地在这3种菌中检测到多态性分子标记.共扩增出了59个DNA片段,其中3个菌株共有的谱带有7条,而显示多态性的片段有52条,占88.1%.引物P6的扩增图谱可用于3株细菌的鉴别.在对3株细菌扩增中发现,鸡鲍氏志贺菌与人痢疾志贺菌的扩增条带相同率达67%,但各有自己的特征性谱带.鸡沙门菌与志贺菌的扩增谱带相差甚远.

The genetic diversity among strains of chicken Shigella boydii, Shigella dysenteriae and S. Pullorum was studied by random amplified polymorphic DNA （RAPD） with 6 random primers. The results showed that four efficient primers were selected from six 10 bp random primers. Fifty-nine RAPD bands were identified and 7 of them were common in all three strains tested and 52 of them were polymorphic between any two strains. The polymorphic bands accounted for 88.1% of the total amplified bands. The primer P6 could be used to identify different species. The result showed that the common rate of the amplified bands between Shigella boydii and Shigella dysenteriae was 67% , but the amplified bands produced by Shigella and S. Pullorum was much diferent from each other when primer P6 was used.