Farber J M, Addison C J
Microbiology Research Division, Health and Welfare Canada, Ottawa, Ontario.
J Appl Bacteriol. 1994 Sep;77(3):242-50. doi: 10.1111/j.1365-2672.1994.tb03070.x.
The randomly amplified polymorphic DNA (RAPD) technique was employed in the development of a typing protocol for Listeria isolates, particularly Listeria monocytogenes strains. A single strain of L. monocytogenes was used and 200 random decamer primers were screened for their discriminatory abilities by visualizing the amplification products electrophoretically. Three candidate primers displaying potentially useful banding patterns were selected and tested against 52 L. monocytogenes strains, encompassing 11 serotypes, and 12 other strains representing five other Listeria spp. Thirty-four banding profiles were obtained with one particular primer. RAPD analysis allowed differentiation between Listeria spp. and was found to further subdivide strains of the same serotype. Where only one primer was used strains from different serotypes were occasionally found to produce identical banding profiles. RAPD analysis, which in our hands proved to be reproducible, shows much promise as a molecular alternative to traditional L. monocytogenes typing protocols.
随机扩增多态性DNA(RAPD)技术被用于开发李斯特菌分离株的分型方案,特别是单核细胞增生李斯特菌菌株。使用了一株单核细胞增生李斯特菌,并通过电泳观察扩增产物来筛选200种随机十聚体引物的鉴别能力。选择了三种显示出潜在有用条带模式的候选引物,并针对52株单核细胞增生李斯特菌菌株(包括11个血清型)和代表其他五种李斯特菌属的12株其他菌株进行测试。使用一种特定引物获得了34种条带图谱。RAPD分析能够区分李斯特菌属,并且发现可以进一步细分相同血清型的菌株。在仅使用一种引物的情况下,偶尔会发现来自不同血清型的菌株产生相同的条带图谱。在我们手中,RAPD分析被证明是可重复的,作为传统单核细胞增生李斯特菌分型方案的分子替代方法,它显示出很大的前景。