本文選題：漢坦病毒 + 腎綜合征出血熱��； 參考：《山東大學(xué)》2005年碩士論文

【摘要】：目的 從山東省嚙齒類等動(dòng)物中鑒定漢坦病毒(HV),陽性標(biāo)本進(jìn)行RT-PCR擴(kuò)增,測定序列,與以往山東省HV病毒株一起進(jìn)行序列分析和分子流行病學(xué)研究。了解它們的同源性和遺傳距離,掌握山東省不同地區(qū)腎綜合征出血熱(HFRS)的分布特征,遺傳規(guī)律,進(jìn)化關(guān)系;找出山東省的代表株,克隆其M基因。為從根本上預(yù)防和控制HFRS在山東省的流行打下堅(jiān)實(shí)基礎(chǔ)并為研制HFRS特異診斷試劑和新型疫苗提供科學(xué)依據(jù)。 方法 采集山東省不同地區(qū)的鼠肺標(biāo)本,應(yīng)用間接免疫熒光法(IFA)鑒定出HV感染的陽性標(biāo)本。根據(jù)GenBank HV M基因保守序列及相關(guān)參考文獻(xiàn)應(yīng)用Primer5軟件設(shè)計(jì)HTN與SEO型通用引物及將HTN型(Ⅰ型)和SEO型(Ⅱ型)分型的型特異性引物。通過RT-PCR法擴(kuò)增陽性標(biāo)本中HV M片段部分基因,測序并用DNASTAR軟件進(jìn)行同源序列比較并構(gòu)建系統(tǒng)發(fā)生樹。觀察山東省HV堿基變異情況,找出山東省的代表株,進(jìn)行全M片段擴(kuò)增并構(gòu)建克隆載體。測序并用BLAST軟件進(jìn)行同源序列比較,觀察核苷酸以及所推導(dǎo)的氨基酸變異情況:利用SignalP、DNASTAR和TmPred軟件預(yù)測其信號(hào)肽序列、疏水區(qū)、抗原區(qū)及跨膜區(qū),分析氨基酸變異對HV包膜糖蛋白生物活性的影響。 結(jié)果 1.捕獲2073只嚙齒目和食蟲目動(dòng)物,鑒定出103份HV感染陽性標(biāo)本,陽性率為4.97%。 2.陽性標(biāo)本經(jīng)RT-PCR擴(kuò)增,得到26份HV相應(yīng)M片段(2003～2302nt)。 3.測序結(jié)果經(jīng)核苷酸序列分析表明26份HV同源性較高在97.3～100.0%,
[Abstract]:Objective to identify Hantavirus (HVV) from rodents and other animals in Shandong Province. The positive samples were amplified by RT-PCR and sequenced. To understand the homology and genetic distance of HFRSs in different regions of Shandong Province, to understand the distribution, genetic law and evolutionary relationship of HFRSs in different regions of Shandong Province, and to find out the representative strains of Shandong Province and clone the M gene of HFRSs. It will lay a solid foundation for the prevention and control of HFRS epidemic in Shandong Province and provide scientific basis for the development of HFRS specific diagnostic reagents and new vaccines. Methods the positive specimens of HV infection were identified by indirect immunofluorescence assay (IFA). According to the conserved sequence of GenBank HV M gene and related references, Primer5 software was used to design HTN and SEO universal primers and type-specific primers to type HTN (type 鈪，

本文編號(hào)：1840293