基于SBT分型技術(shù)的廣州地區(qū)嗜肺軍團(tuán)菌群體遺傳學(xué)研究
發(fā)布時(shí)間:2018-04-30 21:37
本文選題:軍團(tuán)菌 + 群體遺傳學(xué)。 參考:《遼寧大學(xué)》2012年碩士論文
【摘要】:嗜肺軍團(tuán)菌(Legionellapneumophila)廣泛存在于各種水生環(huán)境中,例如江、河、湖泊、冷卻水塔、空調(diào)、噴泉、加濕器等。嗜肺軍團(tuán)菌是軍團(tuán)病的致病因子,吸入軍團(tuán)菌污染的水汽可發(fā)生軍團(tuán)病的感染,經(jīng)常引起散發(fā)病例,以及社區(qū)獲得的暴發(fā)疫情。本研究針對(duì)廣州地區(qū)分離的116株嗜肺軍團(tuán)菌,采用序列分型方法(Sequence-based-technology,SBT)方法進(jìn)行分型研究,旨在了解廣州地區(qū)水環(huán)境中嗜肺軍團(tuán)菌的群體遺傳學(xué)特征,為我國(guó)軍團(tuán)病的分子流行病學(xué)研究提供依據(jù)。對(duì)116分離株的7個(gè)等位基因進(jìn)行PCR擴(kuò)增測(cè)序,獲得各個(gè)分離株的等位基因譜(allelicprofile)和序列型(sequencetype,ST)。隨后以等位基因譜和ST型為基礎(chǔ),利用Bionumerics6.6,eBURST,START2.0,SplitsTree4.0,RDP3.0,ClonalFrame等軟件,對(duì)廣州地區(qū)軍團(tuán)菌的群體遺傳學(xué)結(jié)構(gòu)進(jìn)行深入分析。Bionumerics6.6軟件對(duì)116分離株SBT分型,共確定了30個(gè)ST型,其中有16個(gè)為新ST型;eBURST對(duì)ST型的克隆群(ClonalComplex,CCs)分析,共得到2個(gè)克隆群和22個(gè)單一ST型(singletons);在CC1中ST-1被鑒定為祖先。利用START2.0分別對(duì)30個(gè)ST和116分離株計(jì)算連鎖不平衡,116株分離樣品得到ISA=0.6969(p0.001),,30個(gè)ST得到的ISA=0.3259(p0.001);SplitsTree4.0對(duì)30個(gè)ST型和116分離株的拼接序列構(gòu)建分裂樹,均得到網(wǎng)狀樹,以上結(jié)果表明嗜肺軍團(tuán)菌中存在重組。利用RDP3.0對(duì)30個(gè)ST型和116分離株的拼接序列進(jìn)行重組事件估算,30個(gè)ST型預(yù)測(cè)到5個(gè)重組事件,而116分離株預(yù)測(cè)3個(gè)可能的重組事件。ClonalFrame計(jì)算重組與突變的比例r/m=0.9(0.95%[0.2-1.9]);表明嗜肺軍團(tuán)菌中雖然存在重組,但不足以影響其群體遺傳學(xué)結(jié)構(gòu)。本研究還結(jié)合歐洲軍團(tuán)菌感染工作組(EuropeanWorkingGroupforLegionellaInfections,EWGLI)對(duì)全世界軍團(tuán)菌的研究數(shù)據(jù),初步分析了世界軍團(tuán)菌的種群結(jié)構(gòu)。綜合以上分析可以確定廣州水環(huán)境分離的116分離株的種群結(jié)構(gòu)為克隆化(Clonal)。
[Abstract]:This study is aimed to understand the population genetics characteristics of basophilic bacteria in the water environment of Guangzhou , and to provide the basis for the molecular epidemiology study of the disease in Guangzhou . Based on the alleles spectrum and ST type , we analyzed the population genetics structure of legionum in Guangzhou area by using software of Bionumerics6 . 6 , eburst , START2 . 0 , SplitsTree 4.0 , RDP3 . 0 , ClonalFrame and so on .
Two clones were obtained and 22 single ST - type ( singletons ) were obtained by eburst ' s analysis of ST - type clones ( CCs ) .
In CC1 , ST - 1 was identified as an ancestor . Using START2.0 , the linkage disequilibrium was calculated for 30 ST and 116 isolates , and 116 isolates were ISA = 0.6969 ( p0.001 ) and 30 ST - derived ISA = 0.3259 ( p0.001 ) ;
By using RDP3.0 to estimate the recombination events , 30 ST - type and 116 isolates were used to estimate the recombination events , while 116 isolates predicted three possible recombination events .
In this study , the population structure of the world military community was preliminarily analyzed . The population structure of 116 isolates from Guangzhou water environment was determined to be Clonal .
【學(xué)位授予單位】:遼寧大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2012
【分類號(hào)】:R378
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 劉洪亮;檢驗(yàn)嗜肺軍團(tuán)桿菌新方法[J];天津科技;2004年02期
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