一种应用 LORF11同源子鉴别诊断鸭瘟病毒的 PCR方法

摘要

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 DEV can be detected. The PCR method could not only detect the infection of duck enteritis virus, but also the origin of the virus.%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4448 bp、3277 bp、934 bp和2518 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。