Reverse transcriptase-polymerase chain reaction to detect avian encephalomyelitis virus.
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.
Xie
Zhiqin
Z
Guangxi Veterinary Research Institute, Nanning, People's Republic of China.
Khan
Mazhar I
MI
Girshick
Theodore
T
Xie
Zhixun
Z
D000818:Animals; D001921:Brain / Q000821:virology*; D002642:Chick Embryo; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D001350:Encephalomyelitis Virus, Avian / Q000235:genetics*; D011233:Precipitin Tests / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D015678:Viral Structural Proteins / Q000235:genetics
D017931:DNA Primers; D015678:Viral Structural Proteins