| 冯云,张海林,杨卫红,章域震,黄丽娟,邓淑珍,孙玉杰,杨杜鹃,周济华.云南省1977-2010年流行性乙型脑炎病毒分子流行病学研究[J].中华流行病学杂志,2016,37(11):1519-1525 |
| 云南省1977-2010年流行性乙型脑炎病毒分子流行病学研究 |
| Molecular epidemiology of Japanese encephalitis viruses isolated in Yunnan province, 1977-2010 |
| 收稿日期:2016-05-20 出版日期:2016-11-10 |
| DOI：10.3760/cma.j.issn.0254-6450.2016.11.017 |
| 中文关键词: 流行性乙型脑炎病毒 E基因 基因型 分子流行病学 |
| 英文关键词: Japanese encephalitis virus E gene Genotype Molecular epidemiology |
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| 中文摘要: |
| 目的 阐明1977-2010年分离自云南的63株流行性乙型脑炎（乙脑）病毒的基因型和分子流行病学特点。方法 病毒株常规接种乳小白鼠，取发病濒死的乳鼠脑组织经研磨制成上清液，提取病毒核酸，通过RT-PCR扩增乙脑病毒E基因序列，采用Clustal X、DNAstar和Mega 5.0等生物学软件进行核苷酸和氨基酸序列分析及系统进化树分析。结果 云南乙脑病毒分离株均可引起乳鼠发病和死亡。经RT-PCR和序列测定获得63株乙脑病毒的E基因序列，进化树和同源性分析表明，47株属基因1型，16株为基因3型。基因1型可分为2个进化群，其中云南最早的基因1型分离株（M28，1977；BN82215，1982）与泰国早期基因1型分离株（U70416，1982；DQ084229，年代不详）同处一个进化群；2007-2010年基因1型分离株分布于两个次级进化分支；16株基因3型分布于3个进化群中，其中20世纪70-90年代分离株分布于两个进化群，2004年分离株处于另一进化群。此外，无论基因1或3型乙脑病毒的抗原性、致病性和毒力等主要氨基酸位点均未发生明显改变。结论 云南省存在基因1和3型乙脑病毒流行，1型为近期主要流行型。云南省不同年代和地域的乙脑病毒分离株存在明显差异并具有遗传多样性特点。本研究还提示基因1型乙脑病毒可能起源于中国云南省及相邻东南亚地区。 |
| 英文摘要: |
| Objective To understand the genetic and molecular epidemiologic characteristics of 63 strains of Japanese encephalitis virus (JEV) isolated in Yunnan province, China during 1977-2010. Methods Suckling mice were inoculated with viruses continuously and the viral nucleic acid were extracted from the brain-grinding supernatants of the infected and moribund mice, then the gene fragments of E region were amplified by RT-PCR. Bioinformatics (Clustal X, DNAstar, Mega 5.0 and other software) was used to analyze the nucleotide and deduced amino acid sequences and phylogenetic trees. Results Yunnan strains of JEV could cause illness and deaths in suckling mice. The results of virus nucleic acid detection and sequencing indicated that nucleotide sequences of E gene of the 63 virus strains were obtained. Phylogenetic tree and homology analyses based on E genomes showed that 47 strains of the experimental virus belonged to genotype 1 (G-1) and 16 strains belonged to genotype 3 (G-3). The 47 isolates of G-1 were divided into 2 clades, of them, the earliest isolates of G-1 (M28, 1977 and BN82215, 1982) in Yunnan of China and the early isolates of G-1 (U70416, 1982; DQ084229, the year is unknown) in Thailand were in one clade, and the isolates of G-1 from 2007-2010 in Yunnan could be divided into 2 subgroups. The 16 isolates of G-3 from Yunnan were divided into 3 clades, among them, the isolates from 1970-1990s in Yunnan were in two clades, and the isolates from 2004 in Yunnan were in one clade. In addition, their main amino acid sites of antigenicity, pathogenic, virulence of both G-1 and G-3 had no significant change. Conclusion JEV G-1 and G-3 co-circulated in Yunnan, and G-1 was predominant. The JEV strains isolated in different years and areas in Yunnan had different molecular epidemiologic characteristics and genetic diversity. The results of this study suggested that JEV G-1 might originate from Yunnan of China and adjacent Southeast Asia region. |
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