16S～23S rDNA间隔区序列PCR和RFLP分析对分枝杆菌复合菌群鉴定的研究

The study of differentiation of several mycobacterium complex by PCR and RELP analysis of 16S-23S rDNA spacer sequence

目的 对结核分枝杆菌复合菌群、鸟 胞内分枝杆菌复合菌群以及龟分枝杆菌龟亚种及脓肿亚种等几种快速生长分枝杆菌进行鉴定。方法 应用通用引物a及结核分枝杆菌特异的引物b ,对受试菌种的 16S～ 2 3SrDNA间隔区序列进行PCR扩增 ,并对引物a扩增产物进行限制性内切酶HaeⅢ、MSPⅠ消化反应。结果 应用引物aPCR扩增及RFLP分析能将除结核分枝杆菌复合菌群外的受试的其它菌群中的各菌种区别开 ;而引物b对受试各菌进行扩增 ,只有结核分枝杆菌有扩增条带出现 ,可将结核分枝杆菌与结核分枝杆菌复合菌群的其它菌区别开 ,达到菌种鉴定目的。结论 应用16S～ 2 3SrDNA间隔区序列PCR和RFLP分析能将结核分枝杆菌复合菌群等几组常规方法难鉴定的菌群加以区别。

Objective To evaluate the value of differentiation of several mycobacterium complex by amplifying 16S-23S rDNA spacer sequence and by restriction enzyme analysis of PCR products at gene level. Method The conservative primer a and special primer b were used to amplify 16S-23S rDNA spacer sequence M. tuberculosis complex, M. avium - M. intracellulare complex and several species of fast-growing mycobacterium such as M. chelonae subsp. chelonae, M. chelonae subsp. abscessus et al. Results The results of PCR and restriction enzyme of PCR products showed that this method was able to differentiate the species tested at species level except M.tuberculosis complex, using special primer b can differentiate M. tuberculosis from other species of the same complex. Conclusion PCR and RFLP analysis of 16S-23S rDNA spacer sequence can differentiate several mycobacterium complex quickly.