两种人体蠕形螨超微结构和基因组提取的研究

发布时间：2018-01-24 19:00

  本文关键词： 蠕形螨 环境电子显微镜(ESEM) 超微结构 DNA提取　出处：《山东大学》2005年硕士论文　论文类型：学位论文

【摘要】：目的 应用环境扫描电子显微镜(environmental scanning electron microscope,ESEM)技术观察人体蠕形螨的超微结构,确定其口器性质,寻找其与致病和分类有关的超微结构。用客观证据说明蠕形螨是否致病,做出超微结构分类并对原形态学分类进行评价。 提取两种人体蠕形螨基因组,为人体蠕形螨分子生物学研究铺平道路。 方法 用刮片式取螨器从成人额部取螨,置于2%洗洁精溶液中,在解剖镜直视下,用自制挑螨器分离清洗干净。 将洁净活螨按观察部位的要求,在解剖镜下摆上电镜样品托,立即扫描,拍摄照片。环境扫描电镜工作参数:工作距离(WD)7.5-9.3mm、高电压(HV)20.0KV、高真空探头(Det)GSED、束斑(Spot)3.0、温度5℃、压力0.5kpa、相对湿度60%。 将洁净新鲜的毛囊蠕形螨(Demodex folliculorum, D. f. )、皮脂蠕形螨(Demodex brevis, D. b. )分别在解剖镜直视下研磨粉碎。用改进的小型昆虫基因组提取方法,分别提取两种人体蠕形螨的基因组DNA,将提取物进行琼脂糖凝胶电泳和紫外分光度计检测。 结果 D. f. 的口器位于颚体腹面。口孔是两片口下板中间的狭长纵向裂隙,有一剑峰状的口针从裂隙中向前下方伸出,但未见牙齿。推断其口器是一种特殊的刺吸式口器。D. b. 的口器与D. f. 相似,但尚未见口针伸出。 D. f. 须肢位于颚体两侧,分三节,末节可向腹面做超过90°的运动。末端似钉鞋底状,其上有7个须肢爪,刺状。前三个呈三角形排列,后四个紧密排成两排。D. b. 须肢较短,末节上只有5个须肢爪。 D. f. 口下板为梨形,外板较平;D. b. 口下板短而宽,呈宽椭圆形,外板唇状。
[Abstract]:Objective to apply environmental scanning electron microscope (ESEM) to environmental scanning electron microscope. ESEM technique was used to observe the ultrastructure of human Demodex, to determine its oral structure, and to search for its ultrastructure related to pathogenicity and classification. Objective evidence was used to explain whether Demodex was pathogenic or not. Ultrastructural classification was made and the original morphological classification was evaluated. Two species of human Demodex genomes were extracted to pave the way for the molecular biology of human Demodex mites. Methods the mites were extracted from the frontal part of adult with scraper and placed in the solution of 2% detergent. The mites were separated and cleaned by self-made mite picker under anatomic microscope. The clean live mites were placed under anatomic microscope and scanned immediately according to the requirements of observation site. The working parameters of environmental scanning electron microscope were as follows: working distance was 7.5-9.3mm. High voltage HVV 20. 0 KV, high vacuum probe Dett GSED, beam spot 3. 0, temperature 5 鈩，

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