马拉色菌表型鉴定及分子生物学鉴定与BIOLOG微生物自动分析系统相关性研究

发布时间：2018-05-22 15:38

本文选题：马拉色菌 + 鉴定　；参考：《复旦大学》2010年硕士论文

【摘要】： 第一部分：马拉色菌属的分类及其在常见皮肤病的分布情况目的：研究马拉色菌属的分类及其在两种常见马拉色菌相关疾病中的分布情况。方法：以标准株作为对照,采用PCR扩增rDNA ITS2区并测序的分子生物学方法将来源于花斑糠疹和马拉色菌毛囊炎的马拉色菌进行分类和描述,分析各菌种在这两种疾病的分布情况。结果：在135例花斑糠疹和马拉色菌毛囊炎分离到6个菌种,共114株马拉色菌。鉴定出球形马拉色菌47株(41%)、合轴马拉色菌40株(35%)、糠秕马拉色菌20株(17.5%)、钝形马拉色菌4株(3.5%)、大和马拉色菌2株(1.8%)、皮炎马拉色菌1株(0.9%)。两种疾病菌种构成与疾病类型有关,近半数花斑癣患者皮损中分离出合轴马拉色菌,大部分马拉色菌毛囊炎中分离出球形马拉色菌。结论：不同疾病马拉色菌菌种构成不同,球形马拉色菌与马拉色菌毛囊炎密切相关；马拉色菌rDNA的ITS2区PCR扩增与测序可用于马拉色菌的分类鉴定。第二部分：利用Biolog微生物自动分析系统建立常见马拉色菌的鉴定数据库目的：利用Biolog微生物自动分析系统建立常见马拉色菌的鉴定数据库,探讨该系统鉴定马拉色菌的应用前景。方法：采用表型及PCR扩增、测序的方法,将临床收集的菌株鉴定至种；选取临床最常见的糠秕、合轴和球形马拉色菌,接种于FF微孔板,记录常见马拉色菌对95种不同碳源的利用情况,描述其各自的生长反应谱,建立鉴定数据库。结果：三种马拉色菌中绝大部分菌株可以利用吐温80、糊精；所有马拉色菌菌株均不能利用D-环式糊精、D-半乳糖醛酸、癸二酸；三种马拉色菌对碳源的利用具有明显的差别,而同种马拉色菌对碳源的利用趋势基本相同。糠秕马拉色菌可以利用N-乙酰-D-葡萄糖胺、D-纤维二糖、i-赤藻糖醇、D-果糖等；而球形和合轴马拉色菌均可以利用D-阿拉伯糖、L-阿拉伯糖、D-核糖和D-木糖；其中合轴马拉色菌可以利用甘油／丙三醇。结论：Biolog微生物鉴定系统采用一种特殊的表型鉴定方法,可以对该三种常见马拉色菌进行鉴定,初步推测该系统在马拉色菌不同菌种鉴定中的应用前景。
[Abstract]:Part I: classification of Malassezia and its distribution in common skin diseases Objective: to study the classification of Malassezia and its distribution in two common malassezia related diseases. Methods: Malassezia derived from pityriasis florescens and Malassezia folliculitis were classified and described by PCR amplification of rDNA ITS2 region and sequencing. The distribution of each strain in the two diseases was analyzed. Results: totally 114 strains of Malassezia were isolated from 135 cases of furfurous plaque and Malassezia folliculitis. A total of 47 strains of Malassezia globosa were identified, 40 strains of Malassezia spp., 20 strains of Malassezia furfura, 4 strains of Malassezia obtuse Malassezia, 2 strains of Malassezia grandis, 1 strain of Malassezia dermatitis, and 1 strain of Malassezia dermatitis were identified. The composition of the two diseases was related to the type of disease. Malassezia sphaeroides was isolated from the skin lesions of nearly half of the patients with tinea versicolor and Malassezia sphaeroides was isolated from most of the malassezia folliculitis. Conclusion: Malassezia sphaeroides is closely related to Malassezia folliculitis and the ITS2 region PCR amplification and sequencing of Malassezia rDNA can be used for classification and identification of Malassezia. Part two: the establishment of identification database of common Malassezia by Biolog microorganism automatic analysis system Objective: to establish a database for the identification of common Malassezia by Biolog microbiological automatic analysis system, and to explore the application prospect of the system in the identification of Malassezia. Methods: phenotypic, PCR amplification and sequencing methods were used to identify the strains collected in clinic, and the most common clinical strains, chaff, rachis and Malassezia globularis, were inoculated on FF microporous plate. The utilization of 95 different carbon sources of Malassezia was recorded, their growth response spectra were described, and the identification database was established. Results: most of the three strains of Malassezia could use Tween 80 and dextrin, all Malassezia strains could not use D-cyclodextrin D-galacturonic acid and sebacic acid. The trend of carbon utilization of Malassezia homogenate was basically the same. Malassezia furfuris can use N- acetyl-D- glucosamine D- fiberdisaccharide-alginate, D- fructose, etc., while D- arabinose L- arabinose D- ribose and D- xylose can be used by Malassezia sphaeraceus, and D- arabinose L- arabinose D- ribose and D- xylose. Malassezia can use glycerol / glycerol. Conclusion A special phenotypic identification method was used to identify the three common Malassezia species in the microbiological identification system. The application prospect of the system in different species identification of Malassezia was preliminarily speculated.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R756

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