本文選題：軍團(tuán)菌　切入點(diǎn)：分類學(xué)鑒定　出處：《中南大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:了解廣東新會(huì)地區(qū)環(huán)境水源中軍團(tuán)菌的分布狀況,對(duì)分離株進(jìn)行分子分型,并檢測其致病性,掌握該地區(qū)環(huán)境水源中軍團(tuán)菌的基本特征,為軍團(tuán)菌病的預(yù)防控制提供依據(jù)。 方法:(1)采取公共場所環(huán)境水源,對(duì)其進(jìn)行軍團(tuán)菌的分離培養(yǎng),并以革蘭染色、生化反應(yīng)、血清學(xué)及mip樣基因測序等方法進(jìn)行菌種鑒定,探討公共場所軍團(tuán)菌分布情況。(2)對(duì)已鑒定的嗜肺軍團(tuán)菌,采取擴(kuò)增片段長度多態(tài)性分析、mip基因序列分析等進(jìn)行分子分型,比較不同分子分型方法的分型能力、重復(fù)性、一致率等,并建立本地區(qū)軍團(tuán)菌流行病學(xué)檔案。(3)應(yīng)用特異性引物檢測已鑒定分型的嗜肺軍團(tuán)菌的lvh和rtxA毒力基因,判斷其致病性;同時(shí)對(duì)其毒力島基因進(jìn)行檢測及基因序列分析,探討不同分型嗜肺軍團(tuán)菌毒力島分布情況及其可能起源。 結(jié)果:(1)8個(gè)采樣點(diǎn)中7個(gè)分離出軍團(tuán)菌,23份水樣中20份軍團(tuán)菌培養(yǎng)呈陽性,陽性率為86.9%,檢出軍團(tuán)菌32株,其中嗜肺軍團(tuán)菌23株,非嗜肺軍團(tuán)菌9株,包括戈氏軍團(tuán)菌5株,菲氏軍團(tuán)菌2株,長灘軍團(tuán)菌2株,同時(shí)發(fā)現(xiàn)同一水樣中有多種軍團(tuán)菌共存現(xiàn)象。(2)24株嗜肺軍團(tuán)菌可分為21個(gè)AFLP型,10個(gè)AFLP群,其中AFLP-A群是新會(huì)地區(qū)優(yōu)勢菌群;采用mip基因序列分析可將該24株嗜肺軍團(tuán)菌分為9個(gè)型,且mip基因測序分子分型與AFLP分型具有相關(guān)性。(3)本實(shí)驗(yàn)室分離的23株嗜肺軍團(tuán)菌,其中強(qiáng)毒株為21株(91.3%),弱毒株為2株(8.7%),未發(fā)現(xiàn)無毒株;攜帶毒力島的菌株為17株(73.91%),且毒力島基因與其他基因起源不一致。 結(jié)論:(1)廣東新會(huì)地區(qū)環(huán)境水中軍團(tuán)菌污染較嚴(yán)重,存在多種軍團(tuán)菌種。(2)該地區(qū)嗜肺軍團(tuán)菌分子分型多樣化,且存在優(yōu)勢菌群。(3)新會(huì)地區(qū)分離的嗜肺軍團(tuán)菌多為強(qiáng)毒株,亦攜帶毒力島,致病性較強(qiáng),已對(duì)人群健康構(gòu)成潛在威脅。(4)應(yīng)用合理的檢測方法,定期對(duì)公共環(huán)境水源進(jìn)行軍團(tuán)菌檢測,及時(shí)發(fā)現(xiàn)問題并采取相關(guān)措施將大大減少軍團(tuán)菌爆發(fā)流行。
[Abstract]:Objective: to understand the distribution of Legionella in the environmental water source of Xinhui area of Guangdong province, to type the isolated strain, to detect its pathogenicity, and to understand the basic characteristics of Legionella in the environmental water source of this area. To provide the basis for the prevention and control of Legionella disease. Methods the isolated and cultured Legionella was isolated from the water source of public places. The bacteria were identified by Gram staining, biochemical reaction, serology and mip like gene sequencing. To investigate the distribution of Legionella in public places, the identification of Legionella pneumophila was carried out by using amplified fragment length polymorphism analysis and sequence analysis of mip gene to compare the typing ability and repeatability of different molecular typing methods. The consistent rate and so on, and established the local Legionella Epidemiology Archives. 3) using specific primers to detect the lvh and rtxA virulence genes of the identified and typed Legionella pneumophila, and to judge the pathogenicity of Legionella pneumophila. At the same time, the virulence island gene of Legionella pneumophila was detected and its gene sequence was analyzed, and the distribution and possible origin of virulence island of Legionella pneumophila were discussed. Results Legionella was isolated from 7 out of 8 sampling sites, 20 of 23 water samples were positive for Legionella, the positive rate was 86.9. 32 strains of Legionella were detected, including 23 strains of Legionella pneumophila and 9 strains of non-Legionella pneumophila, including 5 strains of Legionella goosenii. Two strains of Legionella philii and two strains of Legionella from long Beach were found to coexist in the same water sample. 24 strains of Legionella pneumophila could be divided into 21 AFLP types and 10 AFLP groups, among which AFLP-A group was the dominant group in Xinhui area. The 24 strains of Legionella pneumophila were classified into 9 types by mip gene sequence analysis, and 23 strains of Legionella pneumophila were isolated in our laboratory. Among them, 21 virulent strains were found to be 91.3%, 2 strains were virulent strains, and no virulent strains were found, and 17 strains carrying virulence island were 73.91%, and the origin of virulence island gene was not consistent with other genes. Conclusion (1) Legionella in environmental water of Xinhui area of Guangdong Province is seriously polluted, and there are many Legionella species. 2) the molecular typing of Legionella pneumophila in this area is diversified, and the dominant bacteria group. 3) the isolates of Legionella pneumophila in Xinhui area are mostly virulent strains. It also carries virulence island, which has a strong pathogenicity, and has posed a potential threat to the health of the population. (4) reasonable testing methods are used to detect Legionella in water sources of the public environment on a regular basis. Timely detection of problems and relevant measures will greatly reduce the outbreak of Legionella.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R378

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