西尼羅病毒感染病原體檢測方法與流行病學(xué)調(diào)查研究
發(fā)布時間:2018-05-11 09:44
本文選題:西尼羅病毒 + 生物學(xué)性狀; 參考:《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2005年博士論文
【摘要】:西尼羅病毒(WNV)感染是一種經(jīng)蚊蟲傳播,以鳥類為主要動物宿主的自然疫源性疾病。人被攜帶病毒的蚊蟲叮咬后而感染,表現(xiàn)為西尼羅熱或西尼羅腦炎。該病自1999年傳入美國以來,已連續(xù)肆虐了六個流行季節(jié),流行范圍迅速擴(kuò)大,感染人數(shù)逐年攀升,給美國社會造成了極大恐慌。目前,我國尚缺乏對WNV感染的系統(tǒng)研究。本研究應(yīng)用免疫學(xué)與分子生物學(xué)技術(shù),建立敏感、特異的WNV感染檢測方法,并在我國開展WNV感染的流行病學(xué)調(diào)查。旨在了解WNV在我國的存在情況和潛在媒介蚊種及自然疫源地性質(zhì),為深入開展WNV相關(guān)研究和大規(guī)模疫源地調(diào)查奠定基礎(chǔ)。 為證實引進(jìn)的WNV毒株應(yīng)用于相關(guān)研究的可行性,對病毒的致病性、細(xì)胞敏感性、形態(tài)學(xué)、免疫原性、分子生物學(xué)特征進(jìn)行了探討。結(jié)果表明:給乳鼠腦內(nèi)接種WNV可致全部動物死亡;WNV可使Vero-E6細(xì)胞與C6/36細(xì)胞產(chǎn)生細(xì)胞病變效應(yīng)(CPE),分別以細(xì)胞圓縮和細(xì)胞融合為主要特征;電鏡下所見病毒體為有包膜、直徑約30~50nm的球狀顆粒;滅活病毒可誘導(dǎo)小鼠產(chǎn)生抗WNV抗體;采用RT-PCR對病毒C-prM區(qū)和E區(qū)部分基因片段進(jìn)行擴(kuò)增,在病毒培養(yǎng)液及感染乳鼠腦組織中均檢測到目的核苷酸片段,經(jīng)BLAST程序分析僅與WNV有較高的同源性。 進(jìn)一步,對WNV和乙型腦炎病毒(JEV)的上述生物學(xué)性狀進(jìn)行了比較,結(jié)果顯示:兩種病毒在電鏡下的形態(tài)、大小和對乳鼠的致病性相似;與WNV不同,JEV所致C6/36細(xì)胞CPE以細(xì)胞脫落為主要特征;WNV與JEV之間存在抗原交叉反應(yīng);用黃病毒通用引物可從WNV和JEV感染組織中檢出病毒核酸,而用特異引物僅能擴(kuò)增出相應(yīng)病毒的核苷酸片段。 在WNV基因組C-prM區(qū)和E區(qū)設(shè)計3對特異引物,以病毒培養(yǎng)液為標(biāo)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.
【學(xué)位授予單位】:中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】:博士
【學(xué)位授予年份】:2005
【分類號】:R181.3
【引證文獻(xiàn)】
相關(guān)期刊論文 前1條
1 趙民;陳創(chuàng)夫;盛今良;郭志儒;;新疆地區(qū)蚊類攜帶西尼羅病毒情況調(diào)查研究[J];動物醫(yī)學(xué)進(jìn)展;2008年03期
相關(guān)博士學(xué)位論文 前1條
1 史利軍;西尼羅病毒核酸及抗體檢測體系的建立與評價[D];華中農(nóng)業(yè)大學(xué);2008年
相關(guān)碩士學(xué)位論文 前1條
1 趙民;新疆地區(qū)蚊類攜帶西尼羅病毒、甲病毒調(diào)查及萊姆病血清學(xué)初步調(diào)查[D];石河子大學(xué);2008年
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