Abstract: To identify and differentiate rapidly the cause of clinical diseases, a double RT-PCR was developed. Two sets of specific primers were designed according to the sequences of avian reovirus (ARV) and avian leukosis virus (ALV) in the GenBank. A double polymerase chain reaction was optimized to simultaneously detect two pathogens of ARV and ALV in this article. The double RT-PCR system would amplify a 485 bp fragment for ARV and a 673 bp for ALV simultaneously or separately in the samples,depending on its infection status. But not specific band amplified from other six avian pathogenic viruses.As little as 10 pg of ARV and 10 pg of ALV RNA were detected using gel electrophoresis after double RT-PCR. To evaluate the double RT-PCR, 31 clinical samples were comparatively detected.The data showed that the double RT-PCR method was 100% coincidence with the single RT-PCR, and it could be used for detection and differential diagnosis of these two viruses.

Key words: double RT-PCR; avian reovirus; avian leukosis virus