一株雏鹅呼肠孤病毒的分离与鉴定.pdfVIP

中国动物传染病学报 2015,23(1): 47-50 Chinese Journal of Animal Infectious Diseases ·简报· 一株雏鹅呼肠孤病毒的分离与鉴定 陈红梅，万春和，程龙飞，傅光华，施少华，傅秋玲，张丹青，黄 瑜 （福建省农业科学院畜牧兽医研究所福建省畜禽疫病防治工程技术研究中心，福州 350013 ） 摘 要：2013年福建省某鹅场发生一起以肝脏和脾脏均有灰白坏死点（肝白点）为典型病变特征的急性传染病。本研究利用10 -3.25 日龄非免疫番鸭胚，从病死雏鹅肝脏中分离到1株病毒。病毒分离株没有血凝活性，其ELD50为10 /0.2 mL，可被鹅呼肠孤病毒 高免血清特异性中和。利用鹅呼肠孤病毒特异性检测引物对FJ-06株病毒尿囊液进行RT-PCR检测，可扩增到约572 bp特异性目的 条带。将RT-PCR产物进行克隆测序，Blast分析表明，克隆的基因片段为鹅呼肠孤病毒δC蛋白基因片段。结果表明成功分离到 1株鹅呼肠孤病毒。 关键词：鹅呼肠孤病毒；分离；鉴定 中图分类号：S852.659.4 文献标志码：B 文章编号：1674-6422 （2015 ）01-0047-04 ISOLATION AND IDENTIFICATION OF A GOSLING REOVIRUS CHEN Hong-mei, WAN Chun-he, CHENG Long-fei, FU Guang-hua, SHI Shao-hua, FU Qiu-ling, ZHANG Dan-qing, HUANG Yu (Fujian Animal Disease Control Technology Development Center, Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Science, Fuzhou 350013, China) Abstract: An acute infectious disease occurred in a gosling fl ock in 2013 in Fujian Province. Gray necrotic spots were observed in livers and spleens. Livers collected from dead goslings were homologized and inoculated into 10-day-old Muscovy duck embryos for virus isolation. The isolated virus had no hemagglutinating activity for chicken and guinea pig red blood cells. When duck embryos were inoculated with the isolated virus, the ELD50 was determined to be 10-3.25/0.2 mL. In addition, the isolated virus was neutralized with the hyper-immune serum from vaccinated geese. Viral RNA was amplifi ed from the isolated virus using RT-PCR with specifi c target primers. A fragment of 572 bp was obtained, then sequenced and analyzed. The results confi rmed that the isolated virus was a reovirus.