本文選題：馬拉色菌屬　切入點(diǎn)：微流芯片　出處：《復(fù)旦大學(xué)》2012年碩士論文

【摘要】：第一部分：馬拉色菌屬的分類及其在常見皮膚病的分布情況 目的：研究馬拉色菌屬的分類及其在二種常見馬拉色菌相關(guān)疾病中的分布情況。 方法：以標(biāo)準(zhǔn)株作對(duì)照,采用PCR擴(kuò)增rDNAITS2區(qū)并測(cè)序的分子生物學(xué)方法,將來源于花斑糠疹和馬拉色菌毛囊炎的馬拉色菌進(jìn)行分類鑒定,分析各菌種在這二種疾病的分布情況。 結(jié)果：從72例花斑糠疹和11例馬拉色菌毛囊炎患者皮損處分離到7個(gè)菌種,共83株馬拉色菌。鑒定出合軸馬拉色菌41株(49.40%)、糠秕馬拉色菌26株(31.33%)、球形馬拉色菌10株(12.05%)、日本馬拉色菌2株(2.41%)、限制馬拉色菌1株(1.20%)、大和馬拉色菌1株(1.20%)、皮炎馬拉色菌1株(1.20%)及合軸與糠秕的混合感染1株(1.20%)。二種疾病菌種構(gòu)成與疾病類型無(wú)明顯關(guān)系,近半數(shù)花斑糠疹患者皮損中分離出合軸馬拉色菌,而馬拉色菌毛囊炎患者皮損中分離出的菌種無(wú)顯著差異。 結(jié)論：不同疾病馬拉色菌菌種構(gòu)成可能不同,合軸馬拉色菌與花斑糠疹密切相關(guān)；馬拉色菌rDNA的ITS2區(qū)PCR擴(kuò)增與測(cè)序可用于馬拉色菌的分類鑒定。 第二部分：利用微流芯片鑒定并分析馬拉色菌屬菌種基因型 目的：探討微流芯片在馬拉色菌鑒定與分型中的應(yīng)用優(yōu)勢(shì)。 方法：采用DNA測(cè)序的方法,將臨床收集的菌株鑒定至種；選取臨床最常見的糠秕、合軸和球形馬拉色菌等7種馬拉色菌分別進(jìn)行RAPD-PCR擴(kuò)增,將PCR產(chǎn)物進(jìn)行微流芯片的菌種基因型鑒定。 結(jié)果：大多數(shù)菌株均可被2種隨機(jī)引物(S22、S24)擴(kuò)增而獲得清晰條帶,但以S22引物擴(kuò)增的條帶更為穩(wěn)定、清晰,作為本研究主要引物。不同種馬拉色菌通過微流芯片基因型定量分析得到不同大小的固定陽(yáng)性條帶,所有菌株均可見種間和種內(nèi)多態(tài)性。使用RAPD結(jié)合微流芯片方法,基本可將8種馬拉色菌(糠秕、合軸、球形、厚皮、斯洛菲、日本、大和及皮膚馬拉色菌)區(qū)分開來。 結(jié)論：RAPD結(jié)合微流芯片方法作為一種快速、高通量、高靈敏性的分析技術(shù),在馬拉色菌種間菌株遺傳多樣性、親緣關(guān)系的分析及新種鑒定中顯示出一定優(yōu)越性,適合用于流行病學(xué)分析。
[Abstract]:Part I: classification of Malassezia and its distribution in common skin diseasesObjective: 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 identified by using PCR amplification of rDNAITS2 region and sequencing, and the distribution of each strain in the two diseases was analyzed.Results: totally 83 strains of Malassezia were isolated from the lesions of 72 cases of furfuria macularis and 11 cases of Malassezia folliculitis.The mixed infection of chaff was 1. 20%.There was no significant relationship between the composition of the two diseases and the type of the disease. Malassezia was isolated from the skin lesions of nearly half of the patients with furfuriasis, but there was no significant difference between the two kinds of bacteria isolated from the skin lesions of the patients with Malassezia folliculitis.Conclusion: the species composition of Malassezia may be different, and Malassezia is closely related to pityriasis flowered, and PCR amplification and sequencing of the ITS2 region of Malassezia rDNA can be used for classification and identification of Malassezia.Part two: identification and analysis of Malassezia genotypes by microfluidic microarrayObjective: to investigate the advantages of microfluidic microarray in the identification and typing of Malassezia.Methods: by DNA sequencing, the collected strains were identified into species, and 7 strains of Malassezia malassezia, including chaff, axon and malassezia, were selected for RAPD-PCR amplification.The PCR products were identified by microfluidic microarray genotyping.Results: most strains could be amplified by two random primers (S22 S24), but the bands amplified with S22 primers were more stable and clear, so they were the main primers in this study.Fixed positive bands of different sizes were obtained by microfluidic microarray genotyping in different stallions, and interspecific and intraspecific polymorphisms were observed in all strains.Using RAPD and microfluidic chip method, eight species of Malassezia (chaff, axon, spherical, thick skin, Slofi, Japan, and Malassezia grandis) can be basically distinguished.Suitable for epidemiological analysis.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R756

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