Yoshikawa Hisao, Wu Zhiliang, Nagano Isao, Takahashi Yuzo
Department of Biological Science, Faculty of Science, Nara Women's University, Kitauoya-Nishimachi, Nara 630-8506, Japan.
J Parasitol. 2003 Jun;89(3):585-94. doi: 10.1645/0022-3395(2003)089[0585:MCSABI]2.0.CO;2.
This study compared specific polymerase chain reaction (PCR) primers and phylogenetic tree analysis of restriction fragment length polymorphism using the small subunit ribosomal RNA gene in various Blastocystis populations. A phylogenetic tree was constructed using 12 restriction enzymes and a sample pool of 22 isolates, including 2 reference strains and Proteromonas lacertae as an outgroup. The analysis showed that the 22 isolates could be separated into 7 clusters. Four of the 7 clusters were mixed groups that comprised isolates from both humans and nonhuman hosts. The other 3 clusters contained isolates from humans or nonhuman hosts only. The phylogenetic analysis also showed that B. hominis isolates from geographical separated areas did not necessarily cluster in the genetically different groups. The results of genetic homology and phylogenetic tree analysis among Blastocystis isolates from humans and animals indicated that all isolates from animals appear to be B. hominis. Polymerase chain reaction amplifications using previously described and newly defined specific primers mirrored the clusters obtained by the phylogenetic tree analysis. Our results show that primer PCR can be used as a powerful tool for the typing of Blastocystis populations.
本研究比较了使用小亚基核糖体RNA基因对不同芽囊原虫群体进行特异性聚合酶链反应(PCR)引物分析以及限制性片段长度多态性的系统发育树分析。使用12种限制性内切酶和一个包含22个分离株的样本库构建了系统发育树,其中包括2个参考菌株以及作为外群的蜥蜴原滴虫。分析表明,这22个分离株可分为7个簇。7个簇中的4个是混合组,包含来自人类和非人类宿主的分离株。其他3个簇仅包含来自人类或非人类宿主的分离株。系统发育分析还表明,来自地理隔离区域的人芽囊原虫分离株不一定聚集在基因不同的组中。人和动物芽囊原虫分离株之间的遗传同源性和系统发育树分析结果表明,所有动物分离株似乎都是人芽囊原虫。使用先前描述的和新定义的特异性引物进行的聚合酶链反应扩增反映了通过系统发育树分析获得的簇。我们的结果表明,引物PCR可作为芽囊原虫群体分型的有力工具。