锦州地区一例蜜蜂残翼病的RT-PCR诊断与蜜蜂残翼病毒的鉴定及同源性分析  

RT-PCR Diagnosis of one strain of deformed wing disease in Jinzhou area and DWV Identification and Homology Analysis

作　　者：张皓淳 张鑫[1,2] 杨作丰[1,2] 马建山[1,2] 邓文超[1,2] 王冰[1,2] Zhang haochun Zhang Xin Yang Zuofeng Ma Jianshan Deng Wenchao Wang Bing(Liaoning provincial animal disease prevention and control center, Shenyang Liaoning 110164 Liaoning provincial institute of animal medical research, Shenyang Liaoning 110164)

机构地区：[1]辽宁省动物疫病预防控制中心,辽宁沈阳110164 [2]辽宁省动物医学研究院,辽宁沈阳110164

出　　处：《现代畜牧兽医》2017年第8期17-22,共6页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：国家自然科学基金(31372435);辽宁省科技厅自然基金(联合基金)(2014022047)

摘　　要：为确诊锦州某蜂场蜜蜂大量死亡病因,本试验采用反转录-聚合酶链反应(RT-PCR)方法,收集病死蜜蜂提取总RNA,进行反转录,获得c DNA。然后,以c DNA为模板,利用检测常见蜜蜂病毒的引物进行PCR扩增。结果表明,从蜜蜂病料中扩增出编码蜜蜂残翅病毒Helicase蛋白酶的基因片段,大小约702 bp,将分离毒株命名为DWV-JZ。测序结果通过BLAST比对以及与Gen Bank中获得的7个序列进行遗传进化分析,证实DWV-JZ与Gen Bank中的DWV-JL1(KP096414.1)同源性达到95%,亲缘关系最近。To diagnose the cause of death of a large number of bee in Jinzhou a apiary. In this study, total RNA was extracted from dead bees, and eDNA were obtained by reverse-transcriptase PCR （RT-PCR） method. Then, using eDNA as a template, PeR amplification was performed using prim- ers for detecting common honeybee viruses. The results show, a gene fragment about 702 bp encoding deformed wing virus helicase protease was amplified from the bee material, the separation of the virus named DWV-JZ. Sequencing results were analyzed by BLAST and proceeded phylogenetie analysis with seven reference sequences obtained in GenBank. It was confirmed that DWV-JZ had 95% ho- mology with DWV-JLl（KP096414.1） in GenBank and had the closest genetic relationship.

关 键 词：蜜蜂残翼病毒(DWV) RT-PCR 诊断 遗传进化分析 

分 类 号：S858.26[农业科学—临床兽医学]