Abstract: We established a rapid detection of avian leukosis virus (ALV) by reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) in this study. According to the published ALV sequences in GenBank, multiple pairs of primers were designed targeting the conserved region of ALV. The primer, reaction system and processes of LAMP were detected and analyzed by the LAMP Real-time Turbidimeter so as to evaluate its specificity and sensitiveness. Meanwhile, the amplified products were colored by SYBR GreenⅠ after completion of the reaction, so that the amplification can be detected with naked eyes and the results were compared to the former detected by LAMP Realtime Turbidimeter. RT-LAMP showed a highly efficient amplification for ALV nucleic acid which performed at 63 ℃ for 36 min. The results detected with naked eyes by the addition of SYBR GreenⅠ at the end of reaction were consistent with the results detected by Turbidimeter. The RT-LAMP assay showed a strong specificity that the results were all negative towards other nosazontology detection in chicken; and it also showed a high sensitivity with a detection limit of 14.2 pg·μL^-1. The results of 24 clinical samples detected by LAMP developed in this study were consistent with the ones tested before by ELISA and COFAL test. A visualized RTLAMP method which can detect ALV rapidly was established and the results can also be read with naked eyes. The method was easy to operation, which was suitable for rapid detection of ALV.

Key words: avian leukosis virus, loop-mediated isothermal amplification (LAMP), detection