摘要
为建立一种鉴别诊断鸭瘟病毒的PCR方法,根据已发表的8株鸭瘟病毒株的长区开放阅读框(LORF11)等位基因序列,在其开放阅读框等位基因的上游5 bp和下游101 bp的位置设计1对特异性引物。应用这对引物对鸭瘟病毒LH2011株、v2085株、VAC株、Clone-03株DNA进行PCR扩增,分别获得了4 448 bp、3 277bp、934 bp和2 518 bp的特异性条带。此PCR方法特异性强,敏感性高,可检测到10TCID50或0.1LD50的鸭瘟病毒。在感染鸭瘟病毒的组织(肝脏)中均能检测到鸭瘟病毒DNA。对疑似鸭瘟的临床病料的检测结果也证明,建立的PCR方法不仅能够鉴别样品是否感染了鸭瘟病毒,而且能鉴别感染的鸭瘟病毒的来源。
To develop a PCR method for differential diagnosis of duck enteritis virus （ DEV） , a pair of PCR primers was designed at 5 bp upstream and 101 bp downstream of its open reading frame based on published LORF11 homologous sequences of 8 duck enteritis virus strains. Using these primers, specific fragments of 4 448 bp, 3 277 bp, 934 bp and 2 518 bp were amplified using the DNAs of strain DEV LH2011, strain v2085, strain VAC and strain Clone-03 as tem-plate, respectively. The PCR method developed is specific to test DEV. Sensitivity test showed that 10TCID50 or 0. 1LD50＆amp;nbsp;DEV can be detected. The PCR method could not only detect the infection of duck enteritis virus, but also the origin of the virus.
出处
《江苏农业学报》
CSCD
北大核心
2014年第5期1071-1076,共6页
Jiangsu Journal of Agricultural Sciences
基金
公益性行业(农业)科研专项基金项目(201303046)
江苏省自然科学基金项目(BK20131334)
江苏省农业科技自主创新项目[CX(12)3061]
