PCR-RFLP分析16s-23srDNA间区序列鉴定分枝杆菌菌种

The value of mycobacterial strains identified by the identification of 16s-23s rDNA spacer region with PCR-RFLP

目的探讨16S～23srDNA间区序列DNAPCR扩增和限制性酶切分析在分枝杆菌分类鉴定中的价值。方法对19种分枝杆菌标准株的16S～23SrDNA间区序列DNA进行PCR扩增并对扩增产物进行限制性内切酶HaeⅢ、MSP1消化反应，分析不同菌种扩增片段及其限制性片段长度多态性的差异。结果，PCR扩增结果显示：分枝杆菌一般扩增出1～2条带，缓慢生长分枝杆菌扩增片段在340～480bP，快速生长分枝杆菌扩增片段集中在470～575bP，单从扩增产物的琼脂糖凝胶电泳只能鉴定33．3％的受试菌种，酶切结果显示：分枝杆菌的酶切图谱彼此不同。结论16S～23SrDNA间区序列DNAPCR扩增和RFLP分析是分枝杆菌分类鉴定的一种快速、有效的方法。

Objective To study the value in the myeobacterial strains identified by the identification of 16s^23s rDNA spacer region with PCR-RFLP. Methods The 16s^23s rDNA spacer regions in 19 species of frequent mycobacterial standard strains were amplified by PCR and the amplified products were subjected to RFLP using the restriction enzymes, Hae Ⅲ, MspⅠ, by which the differences among the amplified fragments and among the restriction fragment length polymorphism （RFLP） were analyzed. Results The results of PCR amplifica- tion showed thatl or 2 bands were always amplified in the mycobacterial strains, that the amplified fragment of slow grower mycobacteria was one between 340 and 480 bp, rapid grower mycobacteria 470 and 575 bp. 33.3% strains could be identified by the amplified products of agarose gel electrophoresis（AGE） and the results digested by the restriction enzymes showed that the macrorestriction maps of the mycobacterial strains were different. Conclusions PCR amplification of the quickly and effective tool 16s^23s rDNA spacer regions s for rapid identification of and RFLP analysis were reliable, mycobacterial strains.