16S-23SrDNA间隔区序列PCR扩增及探针杂交用于偶然分支杆菌暴发感染的研究  

Identification of post-injection outbreak of M.fortuitum by 16S-23S rDNA spacer sequences

作　　者：张灵霞[1] 庄玉辉[1] 池英藩 邱志强[2] 王玮[2] 刘坦业[2] 池细弟 高世华 林蓉金 

机构地区：[1]解放军三○九医院结核研究室 [2]福建省结核病防治所 [3]福建省第一人民医院

出　　处：《中国防痨杂志》2000年第4期206-208,共3页Chinese Journal of Antituberculosis

摘　　要：目的　从基因水平上确认福建省南平市注射后偶然分支杆菌暴发感染的致病菌 ,并建立PCR结合DNA探针杂交快速鉴定偶然分支杆菌的方案。方法　根据偶然分支杆菌 16S 2 3SrDNA间隔区序列 ,设计合成一段寡核苷酸探针 ,采用PCR扩增、RFLP和DNA斑点杂交技术 ,对 2 9株偶然分支杆菌临床分离株进行鉴定。结果　 2 9株偶然分支杆菌临床分离株均被PCR扩增出一条约 4 70bp的DNA条带 ,DNA探针杂交也出现特异的杂交斑点。结论　 16S 2 3SrDNA间隔区序列PCR扩增及DNA探针杂交在基因水平上确认引起此次注射后暴发感染的致病菌为偶然分支杆菌 ,该方法具有灵敏、特异的特点。Objective To determine the pathogen of the post-injection infection outbreak in Nanping at gene level with molecular biological technique and to establish a rapid identification method of M.fortuitum by PCR/RFLP and DNA probe technique.Methods A single pair of prime and oligonucleotide probe of M.fortuitum were designed,according to the sequence of mycobacterium of 16S-23S rDNA spacer sequences.29 clinical strains were amplified by PCR,then dot-blot hybridization and RFLP of PCR products were made.Results 470bp fragment were produced by PCR of 29 clinical strains,three fragment were produced by Hae Ⅲ,MSP Ⅰ digest of PCR products respectively.The probe only hybridized with M.fortuium and 29 clinical strains.Conclusion The results showed that the pathogen of infection outbreak were M.fortuitum.The 16S-23S rDNA PCR investigative system is sensitive and specific,which can identify M.fortuitum at gene level.

关 键 词：偶然分支杆菌 聚合酶链反应 DNA探针 

分 类 号：R521[医药卫生—内科学] R378.91[医药卫生—临床医学]