本文選題：肺孢子蟲 + 分子系統(tǒng)發(fā)育��； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2005年博士論文

【摘要】：目的:以mt LSU rRNA和ITS1-5.8SrRNA-ITS2兩種基因為分子標(biāo)記,構(gòu)建系統(tǒng)發(fā)育樹,探討肺孢子蟲與宿主的進(jìn)化關(guān)系以及屬的分類地位。方法:采用按動物實際體重定量皮下注射地塞米松法誘導(dǎo)實驗動物(SD大鼠、ICR小鼠、長爪沙鼠、新西蘭白兔、C57BL/6N小鼠和豚鼠)感染肺孢子蟲。收集實驗動物肺泡灌洗液、肺組織、臨床疑似病例痰液樣本。提取肺孢子蟲總DNA。設(shè)計合成擴(kuò)增2個目的基因PCR引物,檢驗其特異性和敏感性。擴(kuò)增目的基因,產(chǎn)物直接或克隆測序,分析比較序列的同源性和進(jìn)化距離。結(jié)合GenBank中相關(guān)基因序列構(gòu)建分子系統(tǒng)發(fā)育樹。結(jié)果:從建立的PCP動物模型及臨床病例樣本獲得了肺孢子蟲DNA。獲得了4種宿主該蟲mt LSU rRNA基因5個序列,其中人1和長爪沙鼠肺孢子蟲的該基因序列未見報道。獲得4種宿主該蟲的ITS1-5.8SrRNA-ITS2基因4個序列,其中長爪沙鼠和新西蘭白兔肺孢子蟲的該基因序列未見報道。構(gòu)建了肺孢子蟲系統(tǒng)發(fā)育樹。結(jié)論:按動物實際體重定量皮下注射地塞米松方法建立了低死亡率PCP動物模型。PCR檢測mt LSU rRNA基因法對PCP檢測可行。肺孢子蟲具有遺傳多樣性和宿主特異性,與其宿主之間呈協(xié)同進(jìn)化關(guān)系。長爪沙鼠和新西蘭白兔肺孢子蟲可能是該屬中獨立的“種”。肺孢子蟲屬歸屬于真菌界的子囊菌門。
[Abstract]:Objective: to construct phylogenetic tree by using mt LSU rRNA and ITS1-5.8SrRNA-ITS2 as molecular markers, and to investigate the evolutionary relationship between Pneumocystis and hosts and the taxonomic status of genera.Methods: Pneumocystis were induced by subcutaneous injection of dexamethasone (Dexamethasone) on the basis of actual animal weight in SD rats ICR mice C57BL / 6N mice and guinea pigs.The samples of alveolar lavage fluid, lung tissue and sputum of clinical suspected cases were collected.Extraction of total DNA from Pneumocystis pneumocystis.Two target gene PCR primers were designed and synthesized to test their specificity and sensitivity.The target gene was amplified, the product was sequenced directly or cloned, and the homology and evolutionary distance of the sequence were analyzed and compared.Molecular phylogenetic tree was constructed by combining the sequence of related genes in GenBank.Results: Pneumocystis pneumocystis DNA was obtained from PCP animal model and clinical samples.Five sequences of Mt LSU rRNA gene were obtained from four species of host, of which human 1 and Pneumocystis longhaurus were not reported.Four sequences of ITS1-5.8SrRNA-ITS2 gene were obtained from four host species of Pneumocystis chinensis, including gerbils of gerbils and Pneumocystis rabbits of New Zealand, which were not reported.The phylogenetic tree of Pneumocystis was constructed.Conclusion: the animal model of low mortality PCP was established by quantitative subcutaneous injection of dexamethasone according to the actual body weight of animals. The detection of mt LSU rRNA gene by PCR was feasible for PCP detection.Pneumocystis have genetic diversity and host specificity, and there is a coevolutionary relationship between Pneumocystis and their hosts.The gerbils and Pneumocystis of New Zealand rabbits may be independent "species" of the genus.Pneumocystis of the genus Pneumocystis belonging to the Ascomystis of the Fungi.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R384

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