艾滋病患者马尔尼菲青霉菌rDNA转录间隔区序列分析  

Sequence analysis of ribosomal DNA internal transcribed spacer of clinical isolates of Penicilliummarneffei obtained from acquired immunodeficiency syndrome patients

作　　者：李凌华[1] 胡凤玉[1] 陈万山[1] 宋伟南[1] 邝燕玲[1] 蔡卫平[1] 唐小平[1] 

机构地区：[1]广州市第八人民医院感染科,510060

出　　处：《中华传染病杂志》2012年第11期647-651,共5页Chinese Journal of Infectious Diseases

基　　金：广东省自然基金资助项目（10151006002000000）

摘　　要：目的了解广东省和广西地区AIDS患者马尔尼菲青霉菌（PM）分离株rDNA转录间隔区（ITS）序列的特征和变异。方法AIDS患者PM分离株15株，其中12株来源于广东，3株来源于广西。运用真菌核糖体通用引物ITSl和ITS4，对其rDNAITS进行扩增和测序，测序结果在GenBank核酸序列数据库中进行同源序列搜索，构建NJ系统树。结果15株PM分离株rDNAITS序列长度为578bp，包括18SrRNA、ITSl、5．8SrRNA、ITS2及28SrRNA，与来源于日本和泰国的PM的rDNAITS序列一致性高达97％～100％，同源性100％；11株广东来源PM和1株广西来源PM菌株rDNAITS序列完全相同，1株广东来源PM和2株广西来源PM菌株的rDNAITS序列存在1个碱基差异；分子进化树显示，PM与毛霉菌、稻根霉菌rDNAITS序列的遗传距离最远，与支顶孢菌、组织胞浆菌及青霉属其他菌种如嗜松青霉、棘孢青霉的rDNAITS序列的种间遗传距离较小。结论广东和广西地区AIDS患者PM分离株的rDNAITS序列非常保守，可作为靶基因来设计PM的种特异性引物，用于菌种鉴定。Objective To analyze the characteristics and mutations of the internal transcribed spacer （rDNA ITS） sequences of Penicillium marneffei （PM） isolates obtained from acquired immunodeficiency syndrome （AIDS） patients in different areas of Guangdong Province and Guangxi Zhuang Autonomous Region. Methods Fifteen PM isolates were obtained from AIDS patients, 12 of which were originated from four geographic areas of Guangdong （western Guangdong, eastern Guangdong, northern Guangdong and central Guangdong） and 3 of which came from Guangxi area. Fungal universal primers ITS1 and ITS4 were applied to amplify the rDNA ITS region of the 15 PM isolates. The polymerase chain reaction （PCR） products were purified and sequenced by T clone sequencing. Homologous sequences of the sequencing results were searched in the nucleotide sequence database of GenBank to construct the neighboring NJ phylogenetic tree. Results The rDNA ITS sequences of the 15 PM isolates obtained from AIDS patients were 578 bp in length, including 18S rRNA, ITS1, 5.8S rRNA, ITS2 and 28S rRNA. These rDNA ITS sequences were highly concordant with those PM isolates originated from Japanese and Thailand （97%- 100%）, with the homology of 100%. The rDNA ITS sequences of 11 PM isolates from Guangdong Province and 1 strain fromGuangxi area were identical, while 2 PM isolates from Guangxi area and 1 strain from Guangdong Province had one single base mutation. NJ phylogenetic tree analysis showed that the genetic distances of rDNA ITS sequences between PM and mold fungus, Rhizopus oryzae were the farthest, while those between PM and Acremonium cellulolyticus, Histoplasma capsulatum, and other species of marneffei, such as Penicillium pinophilum, Penicilliurn aculeatum were closer. Conclusions The rDNA ITS sequence of clinical PM isolate is highly conserved in Guangdong and Guangxi areas. Therefore, it may be the target gene to design the species specific primer for strain identification.

关 键 词：马尔尼菲青霉菌 DNA 核糖体 序列分析 获得性免疫缺陷综合征 

分 类 号：R512.91[医药卫生—内科学]