河南省啮齿类动物流行性出血热病毒的分子流行病学研究

发布时间：2018-06-28 12:39

本文选题：汉坦病毒 + 免疫荧光　；参考：《山东大学》2005年硕士论文

【摘要】：目的:采用分子流行病学的方法,通过汉坦病毒核酸RT-PCR分型及部分扩增片段的序列分析,研究河南省不同地区啮齿类动物携带肾综合征出血热病毒状况和病毒型别,了解其基因型的分布、变异、新型别存在与否,并对其系统发生及进化关系加以分系,为肾综合征出血热防治研究提供科学依据。方法:笼捕法捕鼠,计算鼠密度,确定鼠种构成。用直接免疫荧光法检测动物肺组织切片,应用汉坦病毒型特异性引物以RT-PCR方法对抗原检测阳性的鼠肺标本扩增M和S基因片段上的特异核苷酸序列并测序,将扩增片段的核苷酸序列与已知病毒序列进行比较,用PHYLIP软件对得到的序列绘制系统发生树,了解其进化关系,以明确基因亚型及其地理分布情况。结果:在河南省不同生境的7个县市区中共布鼠笼4720个,捕获啮齿类动物695只,褐家鼠为优势鼠种,占33.53%。其次为黄胸鼠,占18.41%。应用直接免疫荧光法检测全部鼠肺,其中阳性肺组织切片95份,阳性率为13.67%。阳性切片中,褐家鼠和仓鼠分别占47.0%和26.32%。阳性标本并用RT—PCR反应扩增并电泳检测。对3份M片段、S片段均扩增阳性标本进行序列测定,并对其核苷酸序列进行分析显示,均为汉城型(SEO)汉坦病毒,与汉坦病毒R22株、L99株、HB55株、Hb86株、SR11株分别属相同亚型。同一地区的汉坦病毒其核苷酸序列同源性较为接近,表明不同亚型间有着明显的地理聚集现象。在研究中无新的型别出现。结论:河南省不同地区出血热病毒的主要宿主动物是褐家鼠,不同宿主动物携带的汉坦病毒之间的核昔酸序列同源性较高,但也存在一定的差异。这种差异形成的原因有待进一步研究。
[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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R181.3

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