北京市褐家鼠种群遗传多态性及与汉坦病毒分布关系的研究  

Study on Association Between Variation of Hantavirus and DNA Genomic RAPD Markers of Different Rattus norvegicus Population from Beijing

作　　者：江佳富[1] 吴晓明[1] 左曙青[1] 王日明[2] 陈立泉[2] 王炳才[3] 敦哲[3] 张泮河[1] 郭天宇[1] 曹务春[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [2]北京市东城区疾病预防控制中心 [3]北京市海淀区疾病预防控制中心

出　　处：《中国媒介生物学及控制杂志》2005年第5期358-363,共6页Chinese Journal of Vector Biology and Control

基　　金：北京市自然科学基金重点资助项目(7021004)

摘　　要：目的了解北京地区褐家鼠不同地理种群遗传多态性与该地区汉坦病毒(HV)基因变异及其分布可能存在的关联.方法选取北京市不同地区捕获褐家鼠,采用巢式RT-PCR法检测HVM基因片段,获得的阳性标本产物进行直接测序,构建系统发育树.另一方面对不同HV来源的宿主个体和其对应种群内随机选取的其他个体,以及其他一些地区鼠类作为参照,用随机扩增多态DNA(RAPD)技术获得褐家鼠基因组DNA多态性图谱,利用RAPDDIST和PHYLIP软件分析褐家鼠种群多态性,并与对应的北京地区的HV差异和分布进行比较.结果选用50个随机引物进行扩增,有5个引物获得清晰、多态性高的RAPD谱带.不同区域种群RAPD标记不尽相同,北京市褐家鼠地理种群之间存在一定程度的遗传分化,大部分种群遗传距离较为接近,其中采集于某农产品集散地的XFD种群遗传距离相对较大,有特异性标记条带.11个HV代表序列均为汉城型汉坦病毒,基因差异为0.1%～8.0%,可分为2个主要的支系,多数毒株变异类型在不同地域中交叉分布,但BjFT01株变异较大,成一个独立的支系.通过Bootstrap分析和系统树图比较,种群分化较大的XFD种群与该区获得1株亲缘关系相对较远的HV相对应.结论北京市不同区域HV及其褐家鼠DNA多态性均存在一定差异,不同种群尤其是输入性种群对北京HV流行影响程度不同,初步显示一些特定种群与特定的基因性别有一定的关联.XFD种群外源基因交流渗入的可能性较大,进一步证明北京市HV随着鼠类种群迁移输入的可能性极大,但仍有待于选用一些特异性标记基因多态性进行继续研究.Objective To understand the association between the genetic variation of hantavirus and genetic differentiation of Rattus norvegicus population from Beijing. Methods The partial M segment of hantaviruses from captured rodent lung samples were amplified and then sequenced. The alignment of the nucleotide and phylogenetic analysis were made with DNASTAR software. RAPD-PCR was used to analyze the DNA polymorphism of R. norvegicus populations. Results The 356 bp M segment obtained from 11 representative samples were all identified as SEOV, with some degree of heterogeneity （0.1%-8.0 % ） between the different geographical locations. Phylogenetic analysis revealed that there exists at least 2 genetical distinct hantaviruses lineages. BjFT01 strain virus represented a mono-phylogentic lineage. 62 clear polymorphic bands received with 4 out of 50 random primers presented the polymorphism. The cluster of analysis showed that the samples can be divided in to different groups. The individual （XFD population） from Fengtai district 2 had a special band and formed a distinct cluster. The inheritic distance of XFD population was larger than between others and was relatively independent group. Comparison of phylogenies inferred from HV and hosts showed the match phylogenies of BjFT01 strain and XFD population. Conclusion RAPID marker reveals the population differentiation of R. norvegicus and has some to extend association with HV variation and distribution at in Beijing. These data suggest that BjFT01 HV and their rodents hosts population share the common differentiation history. It＇s farther approved that Beijing Seoul virus is more probability to be transmitted from other place with the rat hosts.

关 键 词：褐家鼠 遗传多态性 随机扩增多态DNA 汉坦病毒 地理种群 分布关系 随机扩增多态DNA(RAPD) 北京市 基因组DNA多态性 RAPD标记 

分 类 号：Q987[生物学—遗传学] S812[生物学—人类学]