华南农业大学学报2017，Vol.38Issue(2)：22-26,5.DOI:10.7671/j.issn.1001-411X.2017.02.005
异育银鲫呼肠孤病毒的分离与鉴定
Isolation and identification of a reovirus strain from Gibel carp
摘要
Abstract
Objective] To discover a new pathogen of Gibel carp ( Carassius auratus gibelio) , and to pro-vide a theoretical basis for the disease prevention and control .[Method] Gibel carps suspected having hemorrhagic disease were collected from a farm in Jilin Province .Tissues were observed for bacterial infection and parasite contamination .PCR was used to detect Cyprinid herpes virus-2.Artificial infection was conducted , the infection filtrate was vaccinated to the carp epithelial tumor cells ( EPC ) , and the EPC were observed by an electron microscope .The virus genome was analyzed by SDS-PAGE and identi-fied by RT-PCR and sequencing .[Result] Neither bacterial nor parasitic infection was detected . Cyprinid herpes virus-2 specific bands were not detected by PCR .After healthy Gibel carps were infected with tissue filtrate of sick fish , the mortality rate reached 86.7%within seven days .Clear cytopathy was detected after four generations via blind passage .The virus particles were observed by an electron micro-scope after negative staining , the particle was spherical with around 70 nm in diameter and with no enve-lope.The virus was initially determined as a reovirus strain ( temporarily named JL-4) .The SDS-PAGE results showed that JL-4 possessed 11 segments of dsRNA , which was the typical characteristic of Aquare-ovirus genome.Cluster analysis for the sequences of the RT-PCR amplification product showed that S 6 sequences from JL-4 and reovirus HZ08 had 99% similarity, confirming that JL-4 was reovirus.关键词
异育银鲫/呼肠孤病毒/鲤上皮瘤细胞/分离与鉴定/SDS-PAGEKey words
Carassius auratus gibelio/reovirus/epithelial tumor cell/isolation and identification/SDS-PAGE分类
农业科技引用本文复制引用
巴翠玉,张培军,赵福广,杜晓燕,焦雪,李月红..异育银鲫呼肠孤病毒的分离与鉴定[J].华南农业大学学报,2017,38(2):22-26,5.基金项目
国家自然科学基金（30972191）；吉林省农业产业技术体系（201506）；948计划 （）
