利用临床分离株的DNA探针检测鸟分枝杆菌-胞内分枝杆菌复合体的初步研究。
Preliminary studies on the detection ofMycobacterium avium-intracellulare complex using DNA probe from a clinical isolate.
作者信息
Sharma P, Bose M, Mohd I, Bagdi S, Raj H G
机构信息
Department of Biochemistry, Vallabhbhai Patel Chest Institute, University of Delhi, 110007 Delhi.
出版信息
Indian J Clin Biochem. 2000 Jul;15(2):83-7. doi: 10.1007/BF02883733.
Genomic DNA from a clinical isolate ofMycobacterium avium-intracellulare complex was purified and cloned in PBR 322 at the tetracycline resistance site using Bam HI restriction enzyme. A 16 kb cloned fragment was purified, radiolabeled and used as a probe. Genomic DNA isolated from nineteen MAC strains, threeM. tuberculosis strains and oneM. kansasii strain were digested with Eco RI restriction enzyme, Southern blotted and hybridized with the 16 kb cloned and labeled fragment. Twelve MAC strains showed positive hybridization although five strains gave faint signals. Positive hybridization was noted in two out of the threeM. tuberculosis strains, possibly due to shared DNA homology. No signal was received from the singleM. kansasii strain used in this study.
从鸟分枝杆菌-胞内分枝杆菌复合群的临床分离株中提取基因组DNA，使用Bam HI限制性内切酶在四环素抗性位点将其纯化并克隆到PBR 322中。纯化出一个16 kb的克隆片段，进行放射性标记并用作探针。从19株MAC菌株、3株结核分枝杆菌菌株和1株堪萨斯分枝杆菌菌株中分离出的基因组DNA用Eco RI限制性内切酶消化，进行Southern印迹，并用16 kb的克隆和标记片段进行杂交。12株MAC菌株显示阳性杂交，尽管有5株信号较弱。3株结核分枝杆菌菌株中有2株出现阳性杂交，可能是由于存在共享的DNA同源性。本研究中使用的单一堪萨斯分枝杆菌菌株未检测到信号。
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