本文選題：漢坦病毒 + 免疫熒光�。� 參考：《山東大學》2005年碩士論文

【摘要】：目的:采用分子流行病學的方法,通過漢坦病毒核酸RT-PCR分型及部分擴增片段的序列分析,研究河南省不同地區(qū)嚙齒類動物攜帶腎綜合征出血熱病毒狀況和病毒型別,了解其基因型的分布、變異、新型別存在與否,并對其系統(tǒng)發(fā)生及進化關系加以分系,為腎綜合征出血熱防治研究提供科學依據(jù)。 方法:籠捕法捕鼠,計算鼠密度,確定鼠種構成。用直接免疫熒光法檢測動物肺組織切片,應用漢坦病毒型特異性引物以RT-PCR方法對抗原檢測陽性的鼠肺標本擴增M和S基因片段上的特異核苷酸序列并測序,將擴增片段的核苷酸序列與已知病毒序列進行比較,用PHYLIP軟件對得到的序列繪制系統(tǒng)發(fā)生樹,了解其進化關系,以明確基因亞型及其地理分布情況。 結果:在河南省不同生境的7個縣市區(qū)中共布鼠籠4720個,捕獲嚙齒類動物695只,褐家鼠為優(yōu)勢鼠種,占33.53%。其次為黃胸鼠,占18.41%。應用直接免疫熒光法檢測全部鼠肺,其中陽性肺組織切片95份,陽性率為13.67%。陽性切片中,褐家鼠和倉鼠分別占47.0%和26.32%。陽性標本并用RT—PCR反應擴增并電泳檢測。對3份M片段、S片段均擴增陽性標本進行序列測定,并對其核苷酸序列進行分析顯示,均為漢城型(SEO)漢坦病毒,與漢坦病毒R22株、L99株、HB55株、Hb86株、SR11株分別屬相同亞型。同一地區(qū)的漢坦病毒其核苷酸序列同源性較為接近,表明不同亞型間有著明顯的地理聚集現(xiàn)象。在研究中無新的型別出現(xiàn)。 結論:河南省不同地區(qū)出血熱病毒的主要宿主動物是褐家鼠,不同宿主動物攜帶的漢坦病毒之間的核昔酸序列同源性較高,但也存在一定的差異。這種差異形成的原因有待進一步研究。
[Abstract]:Objective: to study the status and type of hemorrhagic fever with renal syndrome (HFRS) carried by rodents in different areas of Henan Province by means of molecular epidemiology, RT-PCR typing of Hantavirus nucleic acid and sequence analysis of some amplified fragments. To understand the distribution, variation and existence of new genotypes, and to divide the phylogeny and evolution of these genotypes to provide scientific basis for the prevention and treatment of hemorrhagic fever with renal syndrome. Methods: to catch mice by cage, calculate the density of mice and determine the composition of rodent species. The specific nucleotide sequences of M and S gene fragments were amplified and sequenced by RT-PCR with specific primers of Hantavirus type. The nucleotide sequence of the amplified fragment was compared with the known virus sequence, and the phylogenetic tree was mapped by using PHYLIP software to understand the evolutionary relationship of the phylogenetic tree, in order to determine the gene subtype and its geographical distribution. Results: there were 4720 rodent cages in 7 counties of different habitats in Henan Province, and 695 rodents were captured. Rattus norvegicus was the dominant rodent species, accounting for 33.53%. The next was Rattus flavipectus, accounting for 18.41%. All the lungs of the rats were detected by direct immunofluorescence assay, 95 of them were positive in lung tissue sections, the positive rate was 13.67%. In the positive sections, Rattus norvegicus and hamster accounted for 47.0% and 26.32% respectively. The positive specimens were amplified by RT-PCR and detected by electrophoresis. Three M fragment S fragments were sequenced and their nucleotide sequences were analyzed. All of them were Seoul type (SEO) Hantavirus and belonged to the same subtype as Hantavirus R22 strain L99 strain HB55 strain Hb86 strain and Hb86 strain respectively. The homology of nucleotide sequence of Hantavirus in the same region was similar, which indicated that there was obvious geographical aggregation among different subtypes. No new types appeared in the study. Conclusion: the main host animal of hemorrhagic fever virus in different areas of Henan Province is Rattus norvegicus. The nucleic acid sequence of Hantavirus carried by different host animals has high homology, but there are some differences. The causes of this difference need to be further studied.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R181.3

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