云南省2株鹿流行性出血热病毒的分离鉴定
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摘要
为了监测云南省反刍动物鹿流行性出血热病毒(epizootic hemorrhagic disease virus,EHDV)的感染情况,本试验在云南省师宗县设立了监控点,筛选出EHDV抗体阴性的10头牛和5只羊作为哨兵动物,每年的5～10月份每周采血1次,11月到次年4月份每月采血1次,通过酶联免疫吸附试验监测其抗体转阳情况,用转阳牛的红细胞接种BHK以分离病毒,用病毒RT-PCR和中和试验鉴定病毒。结果显示,从2头转阳牛的抗凝血中分离到2株EHDV,其TCID_(50)分别为10~(-2.5)/0.1 mL和10~(-3.44)/0.1 mL,血清型均为EHDV-5型。本试验在云南省分离到EHDV,明确了云南省反刍动物存在EHDV感染,为我国监控EHDV流行情况及疫病风险防范提供了重要借鉴意义。
In order to monitor the infection status of epizootic hemorrhagic disease virus(EHDV),a monitory point was set up in Shizong County(Yunnan Province),ten cattles and five goats with negative antibody against EHDV were choozed as sentinel animals,from June to Octerbor blood samples once a week,from November to April blood samples once a month,using ELISA to monitor their antibodies,using positive cattle red blood cells inoculated BHK to isolate virus,virus neutralization test and quantitative PCR to identify virus.Results showed that two EHDV were isolated,TCID_(50) was 10~(-2.5)/0.1 mL and 10~(-3.44)/0.1 mL.Serotype was EHDV-5.This was the first report that EHDV has been isolated in Yunnan Province,which proved the existence of EHDV infection among the ruminants of Yunnan Province,and provided a reference to monitor EHDV epidemic situation and epidemic risk prevention in China.
In order to monitor the infection status of epizootic hemorrhagic disease virus(EHDV),a monitory point was set up in Shizong County(Yunnan Province),ten cattles and five goats with negative antibody against EHDV were choozed as sentinel animals,from June to Octerbor blood samples once a week,from November to April blood samples once a month,using ELISA to monitor their antibodies,using positive cattle red blood cells inoculated BHK to isolate virus,virus neutralization test and quantitative PCR to identify virus.Results showed that two EHDV were isolated,TCID_(50) was 10~(-2.5)/0.1 mL and 10~(-3.44)/0.1 mL.Serotype was EHDV-5.This was the first report that EHDV has been isolated in Yunnan Province,which proved the existence of EHDV infection among the ruminants of Yunnan Province,and provided a reference to monitor EHDV epidemic situation and epidemic risk prevention in China.
引文
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[8] 曹颖颖.广西首例牛源鹿流行性出血热病毒的分离鉴定[J].中国预防兽医学报,2015,37(10):746-750.
[9] 吕敏娜,朱建波.广东1株牛源流行性出血热病毒的分离鉴定[J].中国预防兽医学报,2017,39(1):67-70.
[2] ARADAIB I E,ALI N O M.Current status and future prospects of epizootic haemorrhagic disease of deer:a review[J].Veterinarski Arhiv,2004,74(1):63-83.
[3] 曹颖颖.鹿流行性出血热病毒研究进展[J].中国兽医学报,2017,37(3):571-576.
[4] 刘胜利.动物虫媒病与检验检疫技术[M].北京:科学出版社,2011:314-321.
[5] 国家质量技术监督总局.中华人民共和国国家标准蓝舌病微量血清中和试验及病毒分离和鉴定方法:GB/T18089-2000[S/OL].http://ishare.iask.sina.com.cn/f/14709772.html.
[6] IWATA H,CHUMA T,ROY P.Charactenrization of the genes encoding two of the major capsid proteins of epizootic haemorrhagic disease virus indicates a close genetic relationship to bluetongue virus[J].J Gen Virol,1992,73(4):915-924.
[7] YADIN H,BRENNER J,BUMBROV V,et al.Epizootic hemorrhagic disease virus type 7 infection in cattle in Israel[J].Vet Rec,2008,162(2):53-56.
[8] 曹颖颖.广西首例牛源鹿流行性出血热病毒的分离鉴定[J].中国预防兽医学报,2015,37(10):746-750.
[9] 吕敏娜,朱建波.广东1株牛源流行性出血热病毒的分离鉴定[J].中国预防兽医学报,2017,39(1):67-70.