Hong Y P, Oh S H, La M S, Im K
Department of Parasitology, Yonsei University College of Medicine, Seoul, Korea.
Korean J Parasitol. 1995 Dec;33(4):341-8. doi: 10.3347/kjp.1995.33.4.341.
Genetic status of Acanthamoeba spp. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Acanthamoeba species, 4 Korean isolates of Acanthamoeba sp., and one American isolate of Acanthamoeba sp. were analyzed by RAPD-PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and stained by ethidium bromide. Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbertsoni and other species (i.e., A. hatchetti, A. triangularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hatchetti and A. triangularis was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triangularis). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbertsoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phenogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hatchetti, A. triangularis, and 3 Korean isolates (YM-2, -3, -4), and the other group consists of A. culbertsoni, A. polyphaga, HOV, and YM-5.
基于随机扩增多态性DNA(RAPD)标记分析对棘阿米巴属的基因状况进行了检测。使用任意十聚体引物通过RAPD-PCR分析了4种先前已确定的棘阿米巴物种、4株韩国棘阿米巴属分离株以及1株美国棘阿米巴属分离株。扩增产物通过琼脂糖凝胶电泳进行分离并用溴化乙锭染色。18种引物产生的DNA扩增图谱显示出4个物种之间存在明显差异。其中9种引物还产生了包含一些分离株特异性扩增产物的DNA扩增图谱。基于18种引物的扩增片段,culbertsoni棘阿米巴与其他物种(即hatchetti棘阿米巴、triangularis棘阿米巴、polyphaga棘阿米巴)之间的成对相似性指数分别为0.300、0.308和0.313。hatchetti棘阿米巴与triangularis棘阿米巴之间的相似性指数为0.833。3株韩国分离株(YM-2、-3、-4)之间的平均相似性指数为0.959,它们与另外2个物种(hatchetti棘阿米巴和triangularis棘阿米巴)之间的平均相似性指数为0.832。YM-5与其他韩国分离株(YM-2、-3、-4)之间的平均相似性指数为0.237。然而,YM-5与culbertsoni棘阿米巴之间的相似性指数为0.857,这表明YM-5在基因上与culbertsoni棘阿米巴比其他韩国分离株更相似。通过UPGMA方法重建的系统发育树显示有两个组:一组由hatchetti棘阿米巴、triangularis棘阿米巴和3株韩国分离株(YM-2、-3、-4)组成,另一组由culbertsoni棘阿米巴、polyphaga棘阿米巴、HOV和YM-5组成。