西尼罗病毒感染病原体检测方法与流行病学调查研究

发布时间：2018-05-11 09:44

本文选题：西尼罗病毒 + 生物学性状　；参考：《中国人民解放军军事医学科学院》2005年博士论文

【摘要】：西尼罗病毒(WNV)感染是一种经蚊虫传播,以鸟类为主要动物宿主的自然疫源性疾病。人被携带病毒的蚊虫叮咬后而感染,表现为西尼罗热或西尼罗脑炎。该病自1999年传入美国以来,已连续肆虐了六个流行季节,流行范围迅速扩大,感染人数逐年攀升,给美国社会造成了极大恐慌。目前,我国尚缺乏对WNV感染的系统研究。本研究应用免疫学与分子生物学技术,建立敏感、特异的WNV感染检测方法,并在我国开展WNV感染的流行病学调查。旨在了解WNV在我国的存在情况和潜在媒介蚊种及自然疫源地性质,为深入开展WNV相关研究和大规模疫源地调查奠定基础。为证实引进的WNV毒株应用于相关研究的可行性,对病毒的致病性、细胞敏感性、形态学、免疫原性、分子生物学特征进行了探讨。结果表明:给乳鼠脑内接种WNV可致全部动物死亡;WNV可使Vero-E6细胞与C6/36细胞产生细胞病变效应(CPE),分别以细胞圆缩和细胞融合为主要特征;电镜下所见病毒体为有包膜、直径约30～50nm的球状颗粒;灭活病毒可诱导小鼠产生抗WNV抗体;采用RT-PCR对病毒C-prM区和E区部分基因片段进行扩增,在病毒培养液及感染乳鼠脑组织中均检测到目的核苷酸片段,经BLAST程序分析仅与WNV有较高的同源性。进一步,对WNV和乙型脑炎病毒(JEV)的上述生物学性状进行了比较,结果显示:两种病毒在电镜下的形态、大小和对乳鼠的致病性相似;与WNV不同,JEV所致C6/36细胞CPE以细胞脱落为主要特征;WNV与JEV之间存在抗原交叉反应;用黄病毒通用引物可从WNV和JEV感染组织中检出病毒核酸,而用特异引物仅能扩增出相应病毒的核苷酸片段。在WNV基因组C-prM区和E区设计3对特异引物,以病毒培养液为标本,
[Abstract]:West Nile virus (WNVV) infection is a naturally infectious disease transmitted by mosquitoes and mainly hosted by birds. People are infected by mosquito bites with the virus, as shown by sirrofever or West Nile Encephalitis. Since the disease was introduced into the United States in 1999, it has been raging through six epidemic seasons in succession, the epidemic range is expanding rapidly and the number of infected people is rising year by year, which has caused great panic to American society. At present, there is no systematic study on WNV infection in China. In this study, immunological and molecular biological techniques were used to establish a sensitive and specific method for the detection of WNV infection, and an epidemiological investigation of WNV infection was carried out in China. The purpose of this study was to understand the existence of WNV in China and its potential vector species and the nature of natural foci, so as to lay a foundation for the further study of WNV and the investigation of large-scale foci. The pathogenicity, cell sensitivity, morphology, immunogenicity and molecular biological characteristics of the imported WNV strains were discussed in this paper. The results showed that inoculation of WNV in the brain could cause all animals to die. WNV could induce cytopathic effect of Vero-E6 cells and C6 / 36 cells, which were mainly characterized by cell shrinkage and cell fusion. Spherical particles about the diameter of 30~50nm; inactivated virus can induce mice to produce anti WNV antibody; RT-PCR was used to amplify some gene fragments of C-prM and E region of the virus. The target nucleotide fragments were detected in the culture medium of virus and the brain tissue of infected mice. The BLAST program only showed high homology with WNV. Furthermore, the above biological characters of WNV and Japanese encephalitis virus (JEV) were compared. The results showed that the morphology, size and pathogenicity of the two viruses under electron microscope were similar to those of young mice. Different from WNV, the CPE of C6 / 36 cells was characterized by exfoliation as the main feature of antigenic cross reaction between WNV and JEV. Nucleic acid could be detected from WNV and JEV infected tissues with yellow virus universal primer. The nucleotide fragments of the corresponding virus could only be amplified by specific primers. Three pairs of specific primers were designed in the C-prM region and E region of WNV genome.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R181.3

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