16S-23S rDNA间隔序列PCR与RFLP对分支杆菌临床分离株的鉴定价值

The value of PCR and RFLP of 16S-233S ribosomal DNA spacerin the differentiation of Mycobacteria clinical isolates

为阐明分支杆菌同种临床分离株不同菌株之间以及与标准株之间１６Ｓ－２３ＳｒＤＮＡ间隔序列是否有差异，以及结核分支杆菌耐药性和１６Ｓ－２３ＳｒＤＮＡ间隔序列有无关系。本文对５８株结核分支杆菌临床分离株（２０株敏感株，３８株耐药株）和淡黄、副偶然、母牛等分支杆菌的临床分离株的１６Ｓ－２３ＳｒＤＮＡ间隔序列进行了扩增，并对扩增产物进行了聚丙烯酰胺凝胶电和琼脂糖凝胶电泳及限制性内切酶ＨａｅⅢ、ＭａｐⅠ消化反应。同种分支杆菌各菌株之间扩增产物及限制性内切酶消化产物均无差异。不同种各株之间均不相同。结果说明同种分支杆菌不同临床分离株１６Ｓ－２３ＳｒＤＮＡ间隔序列没有差异，结核分支杆菌１６Ｓ－２３ＳｒＤＮＡ间隔序列与菌株耐药性没有关系。１６Ｓ－２３ＳｒＤＮＡ间隔序列ＰＣＲ扩增与ＲＦＬＰ分析对分支杆菌临床分离株的鉴定有价值。

To evaluate the possibility of differentiating type of clinical isolates of same Mycobacterium species and to detect whether 16S-23S rDNA relates to drug-resistance M.tuberculosis,We amplified 58 M.tuberculosis strains(20 sensi- tive strains,38 drug-resistance strains)and 12 M.gilvum clinical strains et al.The amplified products as well as the HaeⅢ. Map Ⅰ-digested products of M.tuberculosis were visulized after agarose electrophoresis and PAGE.The amplified fragments and the restriction profiles of the investigated species were different from each other,but no difference were found in the clinical isolates of same species.This study suggested the method can be useful for specise differentiation of Mycobacteria.