Abed Y, Bollet C, de Micco P
Hôpital Salvator, Marseille, France.
Res Microbiol. 1995 Jun;146(5):405-13. doi: 10.1016/0923-2508(96)80286-5.
Amplification of the region separating the genes coding for the two rRNA species 16S and 23S was performed to identify 56 mycobacterial strains, belonging to eleven species: Mycobacterium tuberculosis, M. avium, M. kansasii, M. gordonae, M. abscessus, M. fortuitum, M. xenopi, M. bovis, M. bovis/BCG, M. africanum and M. intracellulare. Reproducible amplification patterns were obtained with most species with the exception of M. kansasii which showed heterogeneity, confirming the existence of a genetically distinct subspecies within this species. In addition, we used the amplified products as target DNA for restriction endonuclease digestion and RAPD (randomly amplified polymorphic DNA) analysis to compare strains of M. abscessus, M. tuberculosis and M. avium. The discriminatory power of these two typing methods was higher than when whole genomic DNA is used as target. Our results demonstrate that the two-step approach to identification and typing on the basis of the hypervariability of 16S-23S spacer region is reliable, rapid and simple, and consequently could be an epidemiological tool in clinical laboratories.
为鉴定56株分枝杆菌菌株，对编码两种rRNA（16S和23S）的基因之间的区域进行了扩增，这些菌株分属于11个种：结核分枝杆菌、鸟分枝杆菌、堪萨斯分枝杆菌、戈登分枝杆菌、脓肿分枝杆菌、偶然分枝杆菌、蟾蜍分枝杆菌、牛分枝杆菌、牛分枝杆菌/卡介苗、非洲分枝杆菌和胞内分枝杆菌。除堪萨斯分枝杆菌表现出异质性外，大多数种均获得了可重复的扩增模式，这证实了该种内存在一个基因上不同的亚种。此外，我们将扩增产物用作限制性内切酶消化和RAPD（随机扩增多态性DNA）分析的靶DNA，以比较脓肿分枝杆菌、结核分枝杆菌和鸟分枝杆菌的菌株。这两种分型方法的鉴别力高于以全基因组DNA作为靶标时。我们的结果表明，基于16S - 23S间隔区高变异性的两步鉴定和分型方法可靠、快速且简单，因此可成为临床实验室的一种流行病学工具。
