应用RT-PCR技术检测番鸭呼肠孤病毒  

Detection of Muscovy Duck Reovirus by RT-PCR

作　　者：胡奇林[1] 林锋强[1] 陈少莺[1] 朱小丽[1] 程晓霞[1] 陈仕龙[1] 林天龙[1] 江斌[1] 李怡英[1] 程由铨[1] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350003

出　　处：《中国兽医学报》2004年第3期231-232,共2页Chinese Journal of Veterinary Science

基　　金：福建省自然科学基金资助项目 ( B0 0 10 0 2 8) ;福建省科技项目 ( 2 0 0 1Z0 61)

摘　　要：参考 Gen Bank中番鸭呼肠孤病毒 (m uscovy duck reovirus,MDRV ) S1基因序列 ,用计算机设计并合成了 1对引物 HP11、HP12 ,以此引物用 RT- PCR对番鸭呼肠孤病毒 S1基因进行了特异性扩增。结果表明 :引物 HP11、HP12能从所有供试的 4株分离毒 MDRV- MW9710、MW980 6、MW980 9、MW9810扩增出 30 0 bp S1基因 c DNA片段 ,而不能从禽呼肠孤病毒 (ARV) S1133株和番鸭胚成纤维 (MDEF)细胞培养物中扩增出任何片段 ;该 RT- PCR的检测灵敏度为 1pg的病毒核酸 ,特异性强 ,重复性好 ,对含毒细胞培养液和尿囊液只需用氯仿进行简单处理 ,即能检测出 MDRV核酸。因此认为 ,该 RT-A pair of primers HP11,HP12 were designed according to the nucleatide sequence of S1 gene of muscovy duck reovirus(MDRV) in GenBank to amplify the S1 gene of MDRV from 4 MDRV strains MDRV-MW9710,MW9806,MW9809 and MW9810 and avian reovirus(ARV) S1133 strain and muscovy duck embryo fibroblast(MDEF) cell cultures.Only one MDRV-specific 300 bp cDNA products were amplified from all the 4 muscovy duck reovirus,but not from ARV S1133 strain and MDEF cell cultures.The detectable viral dsRNA amount of this RT-PCR was 0.01 pg.The results of 3 individual times were consistant.These results indicate that this RT-PCR can be used for rapid detection of the muscovy duck reovirus disease.

关 键 词：RT-PCR 检测 番鸭 呼肠孤病毒 基因序列 

分 类 号：S852.659.4[农业科学—基础兽医学] S858.325.3[农业科学—兽医学]