摘要
为了解阿卡斑病毒在云南师宗的流行情况及病毒的遗传特征,本研究在云南师宗设立哨兵动物,2014年5~11月每周1次连续采集血清、血液样品,血清用做AKAV抗体检测,血液在C6/36、BHK-21、MDBK和Vero细胞上进行病毒分离工作,用S基因特异引物对阳性分离物进行鉴定及序列分析。结果显示,15头哨兵动物共采集到28批420份血清、420份血液样品,其中有3头牛和1只羊感染AKAV,从这些样品中分离获得2株AKAV(编号09312D和09312H),它们在BHK-21和MDBK细胞上48 h产生明显CPE。2株新分离AKAV的S片段序列核苷酸同源性为100%,氨基酸同源性为100%,与所引用的9株其他不同地区分离的AKAV的S片段序列核苷酸同源性在83.8%~97.7%之间,氨基酸同源性在90.6%~99.6%之间,且新分离毒株处于同一进化小分支,具有高度亲缘性,形成了一个独立的次级分支,同时与其他2株中国AKAV分离株处于同一进化簇。结果表明,云南省师宗县为AKAV流行地区,病毒在当地活动频繁,新分离得到的2株病毒与其他AKAV毒株具有较高的核苷酸同源性和氨基酸同源性,提示亚洲太平洋地区AKAV病毒长期流行且关系密切,同时其氨基酸同源性高于核苷酸同源性也提示了AKAV S片段良好的氨基酸遗传稳定性。
To studying the epidemiology and genetic characteristics of Akabane virus from Shizong county in Yunnan province.10 healthy cows and 5 healthy goats which were not infected with Akabane virus were selected to monitor Akabane in Shizong County in 2014.The blood of the selected animals was collected once a week from May to October and once a month in November and December.Use the serum samples and the heparin anticoagulant blood samples to Akabane antibody detection and virus isolation,respectively.The unknown virus was identified by using S gene specific primers.We had collected 420 serum samples and 420 heparin anticoagulant blood samples from 15 sentinel animals.3 cows and 1 goat were infected with Akabane virus,and 2 strains AKAV were isolated in BHK-21 and MDBK cell(No.09312 D and 09312 H).Base on the S segment sequence analysis,09312 D and 09312 H nucleotide sequence homology was 100%,and amino acid homology was 100%,and their S segment nucleotide sequence homology and amino acid homology were 83.8%-97.7%and 90.6%-99.6%respectively between the 10 different geographical regions.What is more,the new separations of two AKAV strains in one small branch,with high affinity in evolution,formed an independent sub branch.At the same time,they were in the same cluster evolution with the other two Chinese AKAV isolates NM/BS/1 and DHL10 M110.The research results showed that Shizong county of Yunnan was a high positive region of AKAV.The activity of AKAV was frequent in this region.There was a close relationship between the two strains AKAV and the other 10 AKAV strains.Their amino acid homology was higher than nucleotide homology,indicating agood amino acid genetic stability of AKAV S segment as well.
作者
孟锦昕
李楠
肖雷
何于雯
寇美玲
宋建领
牛保生
李华春
MENG Jin-xin;LI Nan;XIAO Lei;HE Yu-wen;KOU Mei-ling;SONG Jian-ling;NIU Bao-sheng;LI Hua-chun(Yunnan Tropical and Subtropical Animal Virus Disease Laboratory,Yunnan Animal Science and Veterinary Institute,Kunming650224,China;Animal Disease Control Center of Shizong County,Shizong,Yunnan 655700,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2019年第9期1703-1709,共7页
Chinese Journal of Veterinary Science
基金
云南省应用基础研究计划(青年基金)资助项目(2015FD065)
公益性行业(农业)科研专项资助项目(201303035)
关键词
阿卡斑病毒
病毒分离
S基因
序列分析
Akabane disease virus
isolation of virus
S gene
sequence analysis
作者简介
孟锦昕(1989-),女,助理研究员,硕士;通讯作者:李华春,E-mail:li_huachun@hotmail.com
