Trichophyton mentagrophytes complex

鬚癬毛癬菌複合種屬於皮癬菌，為重要的致病真菌，在臨床上可造成各樣的皮膚感染，如頭癬、臉癬、體癬、手癬、足癬、甲癬等。其宿主範圍極廣，可寄生於人類、貓、狗、鼠、刺蝟身上，利用人類及動物身上的角質物質為營養來源。傳染途徑為人與人、及人與動物直接或間接的接觸造成感染，為公共衛生上一個重要的問題。因此，正確的真菌培養及檢定，有助於了解感染的來源，並加以阻斷，使病人得到完全的治療。癬毛癬菌複合種傳統上包括三個主要的種：鬚癬毛癬菌、趾間毛癬菌、及刺蝟毛癬菌。由於三者在形態學上相似，且存在著許多的中間型及變異型，而使此一複合種成員的命名產生相當大的混淆，同時使三者的分類地位無法完全釐清。這不僅造成在研究時我們可能會選到錯誤的菌株，也使得菌種統計數據變得不可靠，阻礙了對此菌的研究。實驗利用台灣本土人類及動物分離出的48株鬚癬毛癬菌複合種菌株，進行研究。除了分析臨床表現與菌種形態的關係外，我們並根據Gräser等人最新的分類方法，使用ITS1-5.8S-ITS2區段及β-tubulin基因及進行分類分析。結果發現，依最新的分類方法，48株鬚癬毛癬菌複合種中，21株屬於趾間毛癬菌人類株分群，22株屬於趾間毛癬菌動物株分群，5株為刺蝟毛癬菌，沒有鬚癬毛癬菌。由於這種分類方法與我們傳統使用的習慣不同，會造成檢定上的混淆，因此我們認為此分類方法的適用性仍待進一步的評估。Trichophyton mentagrophytes complex, belonging to dermatophytes, is a group of important pathogenic fungi that can cause various human skin infections such as tinea capitis, tinea faciei, tinea corporis, tinea manuum, tinea pedis and onychomycosis. They have a wide range of hosts, including human, cats, dogs, rats, and hedgehogs and can use the keratinous substracts from the hosts as the nutrition source. This is an important public health issue because they can transmit between humans and between human and animals by direct or indirect contact. Culturing and accurate identification of these fungi can help to clarify the routes of infection. This is also very important in the treatment of patients.raditionally, T. mentagrophytes complex consists of three main species, viz. T. mentagrophytes, T. interdigitale and T. erinacei. Due to morphological similarity and existing intermediate forms and variants, there is a great confusion in their taxonomic status. This confusion leads to trouble in choosing correct strains for research and the epidemiological statistics of fungal infections by this group of fungi and further impeding our communicating about it.n this study, we used 48 isolates of the T. mentagrophytes complex obtained from human and animals in Taiwan to perform clinical analysis and molecular taxonomy. We examined the correlation between morphology of the isolates and the clinical presentations. We also used ITS1-5.8S-ITS2 andβ-tubulin gene regions for molecular taxonomy according to the latest taxonomic definition of the T. mentagrophytes complex proposed by Gräser et. al. By this definition, our result shows that among 48 isolates, 21 isolates belong to the anthropophilic strain of T. interdigitale; 22 isolates belong to the zoophilic strain of T. interdigitale; and 5 isolates belong to T. erinacei. No T. mentagrophytes was found. Because the new classification is quite different from what it used to be, confusion in the nomenclature of the T. mentagrophytes complex will be expected. The applicability of Gräser’s taxonomic concept requires further evaluation.口試委員會審定書 i家圖書館碩博士論文授權書 ii謝 iii錄 iv文摘要 vibstract vii一章 緒論 1.1. 真菌感染症 1.2. 真菌培養及檢定在診斷及治療皮膚真菌感染症的重要性 1.3. Trichophyton mentagrophytes comlex簡介 2.3.1. Trichophyton mentagrophytes (C.P. Robin) R. Blanchard. 2.3.2. Trichophyton interdigitale Priestley 3.3.3. Trichophyton erinacei (J.M.B. Sm. & Marple) Quaife 3.4. Trichophyton mentagrophytes complex分類 4.4.1. 形態學分類 4.4.2. 生物種的分類(Taxonomy by biological species) 5.4.3. 分子分類 6二章 研究方法與材料 9.1. 菌株分離 9.1.1. 人類分離株 9.1.2. 動物分離株 9.2. 形態學檢定 10.2.1. Trichophyton mentagrophytes 10.2.2. Trichophyton interdigitale 10.2.3. Trichophyton erinacei 10.3. 分子研究 11.3.1. 培養菌絲 11.3.2. 萃取DNA 11.3.3. 聚合酶鏈結反應(polymerase chain reaction, PCR) 11.3.4. 純化 12.3.5. 定序 13.3.6. 分子分類分析 13三章 結果 14.1. 菌株分離 14.1.1. 人體分離株(表一) 14.1.2. 動物分離株(表二) 15.2. 形態學檢定 16.2.1. 菌落形態 16.2.2. 菌株顯微構造 17.3. 分子研究 17.3.1. ITS1-5.8S-ITS2區段(圖2.3.1) 18.3.2. β-tubulin基因(圖2.3.2) 19四章 討論 20.1. 臨床表現 20.2. 菌種形態分析 21.3. 分子分類結果 21.4. 特殊菌種討論 22.4.1. MCCF 74.08天竺鼠分離株 22.4.2. MCCF 39.05 人類手癬分離株 22.4.3. 手癬分離株 23.4.4. Trichophyton mentagrophytes新模式菌種CBS 318.56 23.5. 對現行檢定法的衝擊 25望 27表 28一、人類來源菌株資料表 28二、動物來源菌株資料表 29三、人類Trichophyton interdigitale人類株分群患者資料 30四、人類Trichophyton interdigitale動物株分群患者資料 312.3.1 Consensus tree of maximal parsimony according to ITS1-5.8S-ITS2 regions of rDNA 322.3.2 Consensus tree of maximal parsimony according to β-tubulin gene 334.3.1. Alignment of ITS1-5.8S-ITS2 regions of anthropophilic and zoophilic strains of T. interdigitale 344.3.2. Alignment of β-tubulin gene of anthropophilic and zoophilic strains of T. interdigitale 354.4.4. Consensus tree of maximal parsimony according to ITS1-5.8S-ITS2 regions of rDNA 36考文獻 37錄 4