云南大理6地域带绦虫的分类研究

发布时间：2019-05-24 06:29

【摘要】：目的：对采自云南大理3县市6地域的人体带绦虫标本,运用形态学及分子生物学方法,鉴定该地区的带绦虫种类,为亚洲带绦虫具体的分类定位提供参考依据,并为云南省带绦虫病和囊虫病的防治提供科学依据。方法：(1)形态学鉴定：选取云南大理3县市6地域带绦虫成虫的头节、成节和孕节节片,采用盐酸卡红和孔雀绿溶液进行染色,与猪带绦虫和传统牛带绦虫比较,对云南大理带绦虫的种类进行初步的形态学鉴定。(2)分子生物学鉴定：从云南大理3县市6地域采集人体带绦虫标本株成虫节片,抽提虫体基因组DNA, PCR扩增线粒体DNA细胞色素B (mtDNA-Cytb)的部分序列,并测序；结合GenBank中已知的猪带绦虫、牛带绦虫和亚洲带绦虫mtDNA-Cytb序列,应用DNAMAN (Version 5.2.2)软件对各标本测序结果进行同源性分析,计算遗传距离并构建系统发育树状图。结果：(1)由形态学分析结果显示,云南大理洱源、大理市双廊和挖色地区带绦虫标本的头节均略呈方形,具有四个吸盘,未见顶突和小钩；成节卵巢分两叶,即左右两叶；孕节的子宫分支大致对称,每侧约13～20支。与报道的亚洲带绦虫相似。(2)根据PCR扩增后mtDNA-Cytb序列测序结果,大理洱源、大理市双廊和挖色地区的带绦虫标本与GenBank中的亚洲带绦虫序列基本一致,同源性分别为99%和100%；大理鹤庆地区带绦虫标本与猪带绦虫同源性为99%。构建的系统发育树显示：EY2、WS及EY1、SL、DY共5个标本株与YZ聚为一支,属于亚洲带绦虫；DL与ND聚为一支,属于牛带绦虫；HQ1-3与ZD聚为一支,属于猪带绦虫；亚洲带绦虫和牛带绦虫两支交汇后再与猪带绦虫汇合。结论：(1)云南大理洱源、云南大理市存在亚洲带绦虫,云南大理鹤庆存在猪带绦虫。猪带绦虫和亚洲带绦虫在云南大理地区有分布区域上的重叠性。(2)mtDNA-Cytb序列分析在鉴定不同种类的带绦虫标本中显示出了有效性。
[Abstract]:Objective: to identify the species of tapeworm in Dali, Yunnan Province by means of morphology and molecular biology, so as to provide reference for the classification and localization of tapeworm in Asia. It also provides scientific basis for the prevention and control of tapeworm and cysticercosis in Yunnan Province. Methods: (1) morphological identification: the head segments, adult segments and pregnant segments of Taenia solium were selected from 6 counties and cities of Dali, Yunnan Province, and stained with kahong hydrochloric acid and malachite green solution, which were compared with Taenia solium and traditional Taenia solium. The species of Taenia solium in Dali belt of Yunnan Province were identified by morphology. (2) Molecular biological identification: adult ganglia tablets of human tapeworm specimens were collected from 3 counties and 6 areas of Dali, Yunnan Province, and the genomic DNA, of Taenia solium was extracted. The partial sequence of mitochondrial DNA cytochrome B (mtDNA-Cytb) was amplified by PCR and sequenced. Based on the known mtDNA-Cytb sequences of Taenia solium, Taenia solium and Taenia solium in GenBank, the homology of the sequencing results of each specimen was analyzed by DNAMAN (Version 5.2.2) software, the genetic distance was calculated and the phylogenetic dendrogram was constructed. Results: (1) the results of morphological analysis showed that the head segments of tapeworm specimens in Dali Eryuan, Dali City and Dali area were slightly square, with four suckers, no apical process and hook, and the ovaries were divided into two leaves, that is, left and right leaves. The uterine branches of the gestational node are approximately symmetrical, with about 13 and 20 branches on each side. It is similar to the reported Asian tapeworm. (2) according to the sequencing results of mtDNA-Cytb sequence after PCR amplification, the tapeworm samples from Dali Eryuan, Dali Shuanglang and Dali area are basically consistent with the Asian Taeniasis sequence in GenBank. The homology was 99% and 100%, respectively. The homology between tapeworm specimens and Taenia solium in Dali Heqing area was 99%. The constructed phylogenetic trees showed that five strains of EY2,WS and EY1,SL,DY were aggregated into one strain with YZ, belonging to Asian tapeworm, one from DL and ND, one from Taenia solium, one from HQ1-3 and ZD, and one from Taenia solium. Asian tapeworm and Taenia solium meet with Taenia solium. Conclusion: (1) Asian tapeworm exists in Dali Eryuan, Yunnan Province, and Taenia solium exists in Dali Heqing, Yunnan Province. The overlap of Taenia solium and Taenia solium in Dali area of Yunnan Province. (2) mtDNA-Cytb sequence analysis showed that it was effective in the identification of different species of tapeworm.
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R383.32

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