本文選題：沙門菌 + 甲型副傷寒沙門菌��； 參考：《中國疾病預防控制中心》2009年碩士論文

【摘要】： 沙門菌血清型復雜,其基因組差異需要深入了解,本研究利用中國沙門菌屬分離株開展多位點序列分型(Multilocus Sequence Typing,MLST)研究,對MLST在不同血清型間及同一血清型內(nèi)亞型的分型能力進行評價,同時評估MLST對鑒定沙門菌屬血清型的參考價值,實驗選擇aroC,dnaN,hemD,hisD,purE,sucA,thrA作為沙門菌分離株MLST的分型基因,對我國參與全球沙門菌監(jiān)測網(wǎng)(Global Salmonella Surveilance,GSS)的5省市2006-2007年分離的201株沙門菌菌株進行分型,并對所有菌株進行血清型鑒定和PFGE實驗,MLST分析將45個血清型沙門菌分離株分為46個序列型(Sequence Type,ST),結(jié)果表明MLST分型結(jié)果與血清型分型結(jié)果有明顯的對應關(guān)系,二者聯(lián)合應用可以作為PFGE實驗結(jié)果的有效補充,沙門菌動物來源和人來源沙門菌的MLST型具有既相互重疊又具有明顯界限的特點。 目前甲型副傷寒沙門菌在我國和東南亞發(fā)病較多,本研究探索建立用于甲型副傷寒沙門菌的多位點可變數(shù)目串聯(lián)重復序列分析(Multiple Locus Variable Numbers Tandem RepeatAnalysis,MLVA)方法并對其應用價值進行評價。從甲型副傷寒沙門菌測序株ATCC9150的全基因組序列中確定所有可變數(shù)目串聯(lián)重復位點(Variable Numbers Tandem Repeat,VNTR)并設計引物,選擇42株在分離時間、地點和PFGE帶型上有差異的甲型副傷寒沙門菌菌株作為panel,從所有VNTR中篩選具有多態(tài)性的位點,利用這些位點建立可用于甲型副傷寒菌株的MLVA實驗方法,再對我國9個地區(qū)1998年-2008年分離的97株甲型副傷寒菌株進行MLVA和PFGE實驗,將這兩種分型方法的實驗結(jié)果進行比較分析,評價MLVA用于甲型副傷寒菌株分型的價值。本實驗建立了基于7個VNTRs位點的甲型副傷寒沙門菌MLVA實驗方法,與PFGE(SpeI酶切)實驗結(jié)果比較,其分型能力要弱于PFGE,但MLVA能夠反映甲型副傷寒沙門菌的流行病學上的差異和關(guān)聯(lián)性,且與PFGE聯(lián)合應用可以提供較PFGE跟高的分辨力,可以作為甲型副傷寒沙門菌分子分型的備選方法。
[Abstract]:The serotype of Salmonella is complex, and the genomic differences need to be deeply understood. In this study, multilocus Sequence typing was carried out by using Chinese Salmonella isolates. The typing ability of MLST between different serotypes and the same serotype was evaluated, and the reference value of MLST in identifying Salmonella serotype was evaluated. 201 strains of Salmonella isolated from 5 provinces and cities of China participating in Global Salmonella Surveillance system (GSS) in 2006-2007 were genotyped. All the strains were identified by serum type identification and PFGE analysis. 45 isolates of Salmonella serotype were divided into 46 serotypes, the results showed that there was a significant correspondence between the MLST typing results and the serotype typing results. The combined application of the two methods can be used as an effective supplement to the results of PFGE experiments. The MLST type of Salmonella from both animal and human sources has the characteristics of overlap and obvious limits. At present, salmonella paratyphoid A is more prevalent in China and Southeast Asia. In this study, a multiple Locus Variable Numbers Tandem repeat analysis (MLVAA) method was established for salmonella paratyphoid A and its application value was evaluated. All variable Numbers Tandem repeats (VNTRs) were identified from the whole genome sequence of ATCC9150, and 42 strains were selected for isolation. As a member of the panel, Salmonella paratyphoid A strains with different loci and PFGE bands were used to screen polymorphic loci from all VNTR, and to establish a MLVA assay for paratyphoid A strains by using these loci. 97 paratyphoid A strains isolated from 9 regions of China from 1998 to 2008 were tested by MLVA and PFGE. The results of the two methods were compared and analyzed to evaluate the value of MLVA in the typing of paratyphoid A strains. In this study, a MLVA assay based on 7 VNTRs loci was established. Compared with the results of PFGE(SpeI digestion, its typing ability was weaker than that of PFGE(SpeI, but MLVA could reflect the epidemiological differences and correlations of Salmonella paratyphoid A. Combined with PFGE can provide higher resolution than PFGE and can be used as an alternative method for molecular typing of Salmonella paratyphoid A.
【學位授予單位】：中國疾病預防控制中心
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R378

【引證文獻】

相關(guān)期刊論文 前1條

1 張東方;袁飛;陳穎;葛毅強;楊海榮;暴書嬋;趙勇勝;;沙門菌分子分型方法研究進展[J];中國公共衛(wèi)生;2012年01期

，

本文編號：1800057