羊及其体表蜱中SFTSV的分离培养与全基因序列分析  

Isolation and analysis of genomic sequences of SFTS virus from goat and tick attached to its surface

作　　者：吴涛[1] 郭喜玲[1] 彭海燕[1] 陈银[1] 赵康辰[1] 崔仑标[1] 葛以跃[1] 史智扬[1] 祁贤[1] 刘大鹏[1] 焦永军[1] 

机构地区：[1]江苏省疾病预防控制中心,卫生部肠道病原微生物重点实验室,南京210009

出　　处：《江苏预防医学》2013年第6期7-10,共4页Jiangsu Journal of Preventive Medicine

基　　金："十二五"科技重大专项计划(2012ZX10004-210-004);江苏省科教兴卫工程(ZX201109;JKRC2011001)

摘　　要：目的 调查2012年春季江苏省某县羊及其体表蜱中发热伴血小板减少综合征病毒（SFTSV）感染情况,并对分离到的毒株进行全基因序列分析.方法 采用荧光定量RT-PCR法对标本进行SFTSV核酸检测,所有阳性标本进行病毒分离并测序,序列结果用Lasergene和MEGA4软件进行拼接和分析.结果 7只羊和从羊体上采集的93只长角血蜱中,1只羊的血清和淋巴结SFTSV核酸检测呈阳性,附着于该羊体表的1只长角血蜱病毒核酸也呈阳性,通过病毒分离获得2株SFTSV毒株,JS2012-yang01（羊）和JS2012-pi01（蜱）.序列分析结果显示,JS2012-yang01和JS2012-pi01的L、M和S基因片段的核苷酸同源性分别为99.8％ 、99.8％ 、100.0％,氨基酸同源性分别为99.6％ 、99.6％、100.0％,进化树分析显示两株病毒处于同一亚分支.2株毒株与中国其他地区流行的SFTSV毒株核苷酸序列同源性介于95.9％～～ 99.6％（L基因）,94.7％～～99.5％（M基因）和95.0％～99.5％（S基因）,与布尼亚病毒科白蛉病毒属其他毒株序列差异较大.结论 该调查中分离自羊和蜱的SFTSV高度同源.蜱虫通过叮咬吸血,可能在SFTSV从动物到人的传播过程中起媒介作用.Objective To investigate the infection of severe fever with thrombocytopenia syndrome virus （SFTSV） in goats and attached ticks in a county of Jiangsu province in spring,2012; to analyze the genomic sequences of the isolated SFTSVs. Methods Real-time RT-PCR was applied to detect SFTSV nucleic acids. Positive samples were cultured for SFTSV virus iso- lation. The complete genomes of the isolated strains were sequenced and analyzed by using Lasergene and MEGA4 software. Results Of the 7 goats and 93 ticks （haemaphysalis longicornis）, the serum and ganglia lymphatica of 1 goat and 1 tick atta- ched to it were confirmed to be SFTSV nucleic acid positive. 2 SFTSV strains （JS2012-yang01 and JS2012-pi01） were isolated, and the homology of nucleotides of the two strains was 99.8%（L）, 99.8%（M） and 100%（S）, respectively; while 99.6%,99. 6 %, 100%for the homology of amino acids, respectively. Phylogenetic analysis demonstrated that they were in the same sub- genotype . The two strains and other representative strains of SFTSVs isolated from other provinces in China shared 95.9 %- 99.6%（L）, 94.7%- 99.5% （M） and 95%- 99.5% （S） for nueleotide acid homology, while they were much more different compared to other isolated bunyaviridae phlebovirus. Conclusion SFTSV viruses isolated from goat and tick are highly homol- ogous . Ticks may play an important role in the transmission of SFTSV from animals to human through biting.

关 键 词：SFTSV 羊 蜱虫 序列分析 

分 类 号：R373[医药卫生—病原生物学]