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目的以ITS1和ITS2基因对中国人源肺孢子菌进行系统发育研究。方法用巢式PCR扩增24例肺孢子菌肺炎(PCP)患者肺泡灌洗液(BALF)中人源肺孢子菌ITS1-5.8S rRNA-ITS2基因,其中包括2株纯培养菌株Pt1和Pt2,分别分离自抗中性粒细胞胞质(ANCA)相关小血管炎合并PCP患者和肾移植合并PCP患者的BALF标本;克隆测序后与GenBank已知序列比对,将多个克隆序列完全相同的标本用于分子系统发育研究。结果进化距离矩阵分析显示,肺孢子菌属内成员间的差异大于克鲁维酵母菌属和曲霉菌各自属内成员间的差异;系统发育树显示,肺孢子菌归属子囊菌门,人和狐猴源肺孢子菌聚为一组,bootstrap值达95%;4种鼠类肺孢子菌聚为一组,bootstrap值达100%。结论肺孢子菌之间的差异是“种”而非“株”的差异,本组比对的肺孢子菌大致分为2组,人和狐猴源肺孢子菌为一组,4种鼠源肺孢子菌为另一组。
Objective To study the phylogenetic relationship of Chinese spirulina with ITS1 and ITS2 genes. Methods The ITS1-5.8S rRNA-ITS2 gene of human Pneumocystis sp. From 24 patients with pneumocystis pneumonia (PCP) was amplified by nested PCR, including 2 strains of pure culture Pt1 and Pt2, The BALF specimens were isolated from anti-neutrophil cytoplasmic (ANCA) -related vasculitis-associated PCP patients and kidney-transplant-associated PCP patients respectively. After cloning and sequencing, they were compared with the known sequences in GenBank and multiple samples with the same clone sequence For molecular phylogenetic studies. Results The evolutionary distance matrix analysis showed that the differences among the members of Pneumocystis were larger than those of members of Kluyveromyces and Aspergillus species. Phylogenetic tree showed that Pneumocystis sp. Pneumocystis sp. Aggregates in one group with a bootstrap value of 95%; the 4 species of Pneumocystis sp. Are clustered together and the bootstrap value is 100%. Conclusions The difference between Pneumocystis is the difference between Pseudomonas aeruginosa and Pseudomonas aeruginosa but not Pseudomonas aeruginosa. There are two groups of Pneumocystis sp. Four species of Pneumocystis sp. Were another group.