de Wit M Y, Klatser P R
Royal Tropical Institute, Department of Biomedical Research, Amsterdam, The Netherlands.
Microbiology (Reading). 1994 Aug;140 ( Pt 8):1983-7. doi: 10.1099/13500872-140-8-1983.
To test for genotypic variations between different isolates of Mycobacterium leprae, the causative agent of leprosy, the 282 bp spacer region between the 16S and 23S rRNA genes was amplified using PCR, and submitted to single-strand conformation polymorphism (SSCP) analysis. The procedure was optimized using four modified spacer fragments, containing mutations at one, three, four and six positions, respectively. Seventy-five M. leprae isolates from different sources, including isolates from leprosy patients, healthy individuals, armadillos and mouse footpads were identical in the SSCP analysis. DNA sequencing and restriction enzyme analysis performed on four and 40 samples, respectively, confirmed the results obtained with SSCP analysis.
为检测麻风病病原体麻风分枝杆菌不同分离株之间的基因型变异,使用聚合酶链反应(PCR)扩增16S和23S rRNA基因之间282 bp的间隔区,并进行单链构象多态性(SSCP)分析。使用四个分别在一个、三个、四个和六个位置含有突变的修饰间隔片段对该程序进行了优化。来自不同来源的75株麻风分枝杆菌分离株,包括来自麻风病患者、健康个体、犰狳和小鼠脚垫的分离株,在SSCP分析中表现相同。分别对四个和40个样本进行的DNA测序和限制性内切酶分析证实了SSCP分析的结果。