Mathieu-Daudé F, Stevens J, Welsh J, Tibayrenc M, McClelland M
California Institute of Biological Research, La Jolla 92037, USA.
Mol Biochem Parasitol. 1995 Jun;72(1-2):89-101. doi: 10.1016/0166-6851(95)00083-d.
Genomic fingerprinting by arbitrarily primed PCR was used to analyze the genetic variability among 59 Trypanosoma brucei stocks representing the three T. brucei subspecies isolated from various hosts and different countries in Africa. 14 oligonucleotide primers revealed 355 polymorphic binary characters which were used for phenetic and phylogenetic analysis and to perform recombination tests exploring the linkage disequilibrium in the sample. There was good concordance between arbitrarily primed PCR polymorphisms and isoenzyme data previously collected for many of the same strains [1]. However, the arbitrarily primed PCR typing was more discerning than multilocus enzyme electrophoresis typing. Phenetic and phylogenetic analysis using arbitrarily primed PCR markers did not confirm T. brucei brucei and T. brucei rhodesiense as separate subspecies, but T. brucei gambiense group I was monophyletic, confirming this group as suitable for the subspecies status. With this exception, there were no clear lineages among the sample, other than clustering of East African stocks and clustering of West African stocks. Some features of the phylogenetic analysis suggested that the population structure was not strictly clonal though recombination tests showed linkage disequilibrium, even in the absence of repeated genotypes. While genotypes appear stable enough for tracking in applied studies, sexuality will impact at the evolutionary time scale, and may be more frequent under some ecological conditions. The arbitrarily primed PCR approach should be an effective and simple approach to follow epidemics and to quantify the role of sexuality in T. brucei populations.
利用任意引物PCR进行基因组指纹分析，以分析59株布氏锥虫株之间的遗传变异性，这些菌株代表了从非洲不同宿主和不同国家分离出的布氏锥虫的三个亚种。14个寡核苷酸引物揭示了355个多态性二元特征，这些特征用于表型和系统发育分析，并进行重组测试以探索样本中的连锁不平衡。任意引物PCR多态性与先前为许多相同菌株收集的同工酶数据之间具有良好的一致性[1]。然而，任意引物PCR分型比多位点酶电泳分型更具辨别力。使用任意引物PCR标记进行的表型和系统发育分析并未证实布氏布氏锥虫和布氏罗德西亚锥虫为独立的亚种，但布氏冈比亚锥虫第一组是单系的，证实该组适合作为亚种地位。除此之外，除了东非菌株的聚类和西非菌株的聚类外，样本中没有明显的谱系。系统发育分析的一些特征表明，种群结构并非严格克隆性的，尽管重组测试显示存在连锁不平衡，即使在没有重复基因型的情况下也是如此。虽然基因型在应用研究中似乎足够稳定以便追踪，但有性生殖将在进化时间尺度上产生影响，并且在某些生态条件下可能更频繁。任意引物PCR方法应该是一种有效且简单的方法，可用于追踪流行病并量化有性生殖在布氏锥虫种群中的作用。
