Fekete A, Bantle J A, Halling S M, Stich R W
USDA/ARS/NADC, Ames, Iowa 50010.
J Bacteriol. 1992 Dec;174(23):7778-83. doi: 10.1128/jb.174.23.7778-7783.1992.
DNA heterogeneity among members of the genus Brucella was demonstrated with the arbitrarily primed polymerase chain reaction (AP-PCR). Simple, reproducible genomic fingerprints from DNA of 25 different Brucella strains were generated with five arbitrarily chosen primers, alone and in pairs, with the PCR. Reaction conditions were optimized for each primer. Several DNA segments were amplified in each sample with all of the primers. PCR products that are not shared among all strains act as polymorphic markers. Polymorphism was apparent for each primer. The Brucella strains can be distinguished according to the banding patterns of their amplified DNA on agarose gels, and the differences can be diagnostic of specific strains. To determine genetic relatedness among the Brucella strains, similarity coefficients were calculated. Statistical analysis of the similarity coefficients revealed the degrees of relatedness among strains of the genus Brucella.
利用随机引物聚合酶链反应(AP-PCR)证明了布鲁氏菌属成员之间的DNA异质性。使用五个随机选择的引物单独或成对进行PCR,从25种不同布鲁氏菌菌株的DNA中生成了简单、可重复的基因组指纹图谱。针对每个引物优化了反应条件。使用所有引物在每个样品中扩增了几个DNA片段。并非所有菌株都共有的PCR产物作为多态性标记。每个引物都表现出多态性。根据布鲁氏菌菌株在琼脂糖凝胶上扩增DNA的条带模式可以区分它们,这些差异可用于诊断特定菌株。为了确定布鲁氏菌菌株之间的遗传相关性,计算了相似系数。对相似系数的统计分析揭示了布鲁氏菌属菌株之间的相关程度。