刘家峡水库IHNV检测与分离鉴定  

Detection,isolation and identification of IHNV in Liujiaxia Reservoir

作　　者：康维国 王建福[1] 吕娜娜 海强 刘哲[1] 成述儒[1] KANG Wei-guo;WANG Jian-fu;LYU Na-na;HAI Qiang;LIU Zhe;CHENG Shu-ru(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070

出　　处：《生物技术》2023年第4期448-452,457,共6页Biotechnology

基　　金：国家自然科学基金项目(32060257);甘肃省农牧厅特种养殖专项(GARS-TZYZ-1);2021年现代丝路寒旱农业科技支撑项目(GSLK-2021-16)。

摘　　要：[目的]为了监测IHNV在刘家峡水库不同水生动物中的流行情况并对其进行鉴定。[方法]利用ELISA抗原检测法对水库上游(三塬电灌区)、中游(刘家村)和下游(祁家渡口)三个主要网箱养殖片区内养殖的虹鳟、金鳟、杂交鲟、青海湖裸鲤、鲤以及野生池沼公鱼和浮游动物开展了IHNV检测;之后用检测阳性的虹鳟的组织样本进行病毒分离、巢式RT-PCR检测,并将扩增产物测序后进行系统进化树分析。[结果]在检测的53份样品中,分别在祁家渡口网箱养殖区1号虹鳟的肾脏、2号虹鳟的肝脏以及2号鲟鱼的肝脏和脾脏混合样中检测出IHNV阳性;将阳性虹鳟的头肾、肝脏、脾脏组织匀浆后接种到EPC细胞中,第5 d开始出现CPE,巢氏RT-PCR检测培养物为IHNV阳性,测得该病毒72 h的滴度为10^(8.23)TCID_(50)/mL,系统进化树分析结果显示,所分离毒株与国内2016年黑龙江报道的HLJ-09株序列同源性最高。[结论]刘家峡水库网箱养殖区的虹鳟、鲟鱼是IHNV的宿主,靠近下游的祁家渡口网箱养殖片区IHNV检出率最高,分离得到的IHNV可能与2016年报道的黑龙江HLJ-09株同源。[Objective]To monitor the prevalence of IHNV in different aquatic animals in Liujiaxia Reservoir and identify it.[Method]IHNV detection was carried out for Oncorhynchus mykiss W.,hybrid sturgeon,Gymnocypris przewalskii and Cyprinus carpio L.,Hypomesus olidus and zooplankton in three main cage culture areas of the reservoir which are upper stream(Sanyuan irrigation area),middle stream(Liujiacun area)and down stream(Qijia ferry area)by ELISA antigen detection method.Then,the positive rainbow trout tissue samples were used for virus isolation,nested RT-PCR detection,and then the amplified products were sequenced for phylogenetic tree analysis.[Result]Among the 53 samples tested,IHNV was detected in kidney of No.1 rainbow trout,liver of No.2 rainbow trout and mixed liver and spleen of No.2 sturgeon in Qijia ferry area,respectively.The head kidney,liver and spleen tissues of positive rainbow trout were homogenized and inoculated into EPC cells,and CPE began to appear on the 5th day.The culture was positive for IHNV by nest RT-PCR.The titer of the virus at 72 hours was 10^(8.23)TCID_(50)/mL.Phylogenetic tree analysis showed that the isolated strain had the highest sequence homology with HLJ-09 strain reported from Heilongjiang in 2016.[Conclusion]The rainbow trout and sturgeon in the cage culture area of Liujiaxia Reservoir are the hosts of IHNV,and the detection rate of IHNV in the cage culture area of Qijiadukou near the downstream is the highest.The isolated IHNV may be homologous to HLJ-09 strain from Heilongjiang Province reported in 2016.

关 键 词：传染性造血器官坏死病 虹鳟 病毒 分离鉴定 检测 

分 类 号：S941.414[农业科学—水产养殖]