Xie Zhiqin, Khan Mazhar I, Girshick Theodore, Xie Zhixun
Guangxi Veterinary Research Institute, Nanning, People's Republic of China.
Avian Dis. 2005 Jun;49(2):227-30. doi: 10.1637/7307-111804R.
A reverse transcriptase-polymerase chain reaction (RT-PCR) was developed and optimized for the detection of avian encephalomyelitis virus (AEV). A pair of primers was prepared based on the VP2 gene of the structural protein P1 region of the AEV genome. An avian encephalomyelitis virus-specific 619-base pair cDNA product was amplified by these primers from five reference/field strains of AEVs but not from 10 other avian pathogenic viruses and bacteria. The RT-PCR assay developed in this study was found to be sensitive and specific with as little as 10 pg of avian encephalomyelitis virus RNA detected using gel electrophoresis. Furthermore, AEV-RT-PCR was able to detect AE virus from chicken embryo brain at 3 days postinoculation as compared with the AE agar gel precipitation test (AGP), which required up to 11 days of incubation in the embryos.
开发并优化了一种用于检测禽脑脊髓炎病毒（AEV）的逆转录聚合酶链反应（RT-PCR）。根据AEV基因组结构蛋白P1区域的VP2基因制备了一对引物。通过这些引物从AEV的五个参考/野外毒株中扩增出一条619个碱基对的禽脑脊髓炎病毒特异性cDNA产物，但从其他10种禽病原病毒和细菌中未扩增出该产物。本研究开发的RT-PCR检测方法被发现具有敏感性和特异性，使用凝胶电泳可检测到低至10 pg的禽脑脊髓炎病毒RNA。此外，与需要在胚胎中培养长达11天的AE琼脂凝胶沉淀试验（AGP）相比，AEV-RT-PCR能够在接种后3天从鸡胚脑中检测到AE病毒。
