环境水军团菌调查及扩增片段长度多态性分型研究

发布时间：2018-04-30 03:33

本文选题：军团菌 + 种属鉴定　；参考：《重庆医科大学》2008年硕士论文

【摘要】： 军团菌(Legionella)是一种兼性细胞内寄生菌,广泛存在于自然界及人工水体环境中。人体吸入污染有军团菌的气溶胶,饮用污染或创面接触污染水源等,都有可能导致军团菌的感染。军团菌属(Legionellace)已发现有50多个种,70多个血清型,并不断有新的菌种从环境中分离。尽管绝大部分的军团菌感染由嗜肺军团菌(L. pneumophila)引起,但鉴于军团菌的原虫宿主细胞内寄生性,在适宜的条件下,可以以相似的方式感染人体巨噬细胞,在这个意义上说,所有的军团菌都具有致病性。近年来,军团菌在西方多个国家引发大的爆发流行,并日益引起人们的重视,目前已有多个国家将军团病(Legionnaire disease,LD)列为法定的传染病。然而,针对军团菌感染的疫苗目前尚未问世,军团病的预防和控制,仍然以输水管道的定期清洗和消毒为主。定期检测人工水体及自然界中军团菌的存在情况,并进行相应的处理,将大大地降低军团病的发病率。我室在参考美国CDC 2005年公布的《环境水军团菌分离培养程序》的基础上,根据军团菌的微生物特性及在存活能力,接种前进行相应的水样预处理,取得了较好的军团菌分离培养效果。2006年8~11月间,于广州市8个公园内采集了44份水样,对广州市地表环境水中的存在情况进行抽样市调查,结果8个公园内均检测到军团菌,阳性率达61.36%。对于初分离的108株军团菌,以AFLP进行同源性分析,得到31株不同菌菌株。经生化反应、血清学鉴定、菌体脂肪酸分析、16S rRNA测序等多种手段,最终鉴定出嗜肺军团菌(L. pneumophila)20株,橡树岭军团菌(L. oakridgensis)1株,圣海伦军团菌(L. sainthelensi)1株,长滩军团菌(L. longbeachae)4株,菲氏军团菌(L. feeleii )5株。参考EWGLI公布的《嗜肺军团菌的AFLP分型草案》1.2版,对广州金域医学检验中心的41株嗜肺军团菌进行分子分型,共得到为31个AFLP型,其中10个型别与EWGLI数据库中的AFLP分型图谱具有可比性。由此可见,采用规范化的AFLP技术,有利于实验室间的资料交流、比对,分子流行病学调查结果也更为有效和可靠。
[Abstract]:Legionella is a facultative intracellular parasite widely found in nature and artificial water environment. Inhaled aerosol contaminated with Legionella, drinking pollution or wound contact with contaminated water may lead to Legionella infection. Legionellace has found more than 50 species and 70 serotypes, and new strains have been isolated from the environment. Although the vast majority of Legionella infections are caused by Legionella pneumophila, given the parasitism of the protozoa host cells of Legionella, it is possible to infect human macrophages in a similar manner under suitable conditions, in that sense, All Legionella are pathogenic. In recent years, Legionnaire disease (LDD) has been listed as a legal infectious disease in many countries. However, the vaccine against Legionella infection is not yet available, and the prevention and control of Legionnaires disease is still focused on the regular cleaning and disinfection of water pipelines. Periodic detection of Legionella in artificial water and nature will greatly reduce the incidence of Legionnaires disease. On the basis of referring to the "procedure of isolation and Culture of Legionella in Environmental Water" published by CDC of the United States of America in 2005, according to the microbial characteristics of Legionella and its survival ability, the corresponding water samples were pretreated before inoculation. 44 water samples were collected from 8 parks in Guangzhou from August to November 2006. The presence of Legionella in surface environmental water in Guangzhou was investigated by sampling city. The results showed that Legionella was detected in 8 parks. The positive rate was 61.36%. For the first 108 strains of Legionella, 31 strains of different strains were obtained by AFLP homology analysis. By biochemical reaction, serological identification and 16s rRNA sequencing, the strains of Legionella pneumophila L. pneumophila)20, Legionella Oaks L. oakridgensis)1, Legionella St. Helene L. sainthelensi)1, Legionella long Beach L. longbeachae)4 were identified. Legionella fernesii L. feeleii 5 strains. According to the draft of AFLP typing of Legionella pneumophila published by EWGLI, the molecular typing of 41 strains of Legionella pneumophila in Jinyu Medical Laboratory Center of Guangzhou was carried out. The results showed that there were 31 AFLP types of Legionella pneumophila. Ten of them were comparable to AFLP genotyping in EWGLI database. It can be seen that the use of standardized AFLP technology is beneficial to the exchange of data among laboratories, and the results of molecular epidemiology investigation are more effective and reliable.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R378

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