雏番鸭细小病毒病病毒分离鉴定  

ISOLATION AND IDENTIFICATION OF A MUSCOVY DUCKLING PARVOVIRUS

作　　者：刘惠莉[1] 李震[1] 王英[1] 张平[1] 丁卫星[2] 韦涛荣[3] 

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海市农业遗传育种重点实验室动物基因工程研究室,上海201106 [2]上海市农业科学院瑞丰科技公司,上海201106 [3]南京农业大学动物医学院,南京210095

出　　处：《上海农业学报》2001年第1期54-58,共5页Acta Agriculturae Shanghai

基　　金：上海市农业科学院青年科技基金项目

摘　　要：采集以腹泻、呼吸困难为主要症状的雏番鸭肝、脾病料 ,接种 14d龄非免疫番鸭胚 ,80 %胚在 3～ 7d死亡 ,胚体呈弥漫性充血、出血。接种番鸭胚成纤维原代细胞 72h可见细胞圆缩、聚团等变化。收集尿囊液及细胞培养物 ,以番鸭细小病毒 (Muscovyducklingparvovirus ,MDPV)阳性血清进行聚苯乙烯乳胶凝集 ,病料培养物均呈阳性反应 ,提取病料接种的尿囊液及细胞基因组DNA ,用自行设计引物PCR扩增到与设计相符的 60 0bp片段 ,经酶切鉴定 ,结果表明克隆到的片段为雏番鸭细小病毒特异性片段 。A kind of virus was isolated from the liver and spleen of Muscovy ducklings with symptoms of diarrhea and breathing with difficulty. Eighty percent Muscovy duckling embryos inoculated with the virus we gain were died in 3～7 days. Muscovy duck embryo fibroblast cells(MDEF) and allantoic liquid infected with virus were tested with Polystyrene LPA and were positive. Furthermore we extracted the genome DNA with proteinase K and SDS. Using a pair of specific primers, we acquire a 600bp DNA fragment, which conincided with what we design. And MDEF was negative in both Polystyrene LPA & PCR test. Furthermore we cloned the gene onto T vector. By digestion with Pst1 and HindIII, the result coincided with what we know for the sequence of MDPV. Thus by polystyrene LPA and PCR test, we can assume that the virus we gain is Muscovy duckling parvovirus(MDPV). Furthermore we have established the method for MDPV diagnosis.

关 键 词：雏番鸭细小病毒病 PCR 病毒 分离鉴定 

分 类 号：S858.325.3[农业科学—临床兽医学] S852.659.2[农业科学—兽医学]