关键词: 柑橘, 柑橘叶斑驳病毒, 环介导等温核酸扩增技术

Abstract: This study aim of is to establishe the detection method of reverse transcription loop-mediated isothermal amplification（RT-LAMP）method for visual detection of CLBV，and validate its sensitivity and accuracy. The primers were designed and screened based on the conserved region of coat protein gene of CLBV. The reaction conditions of the RT-LAMP，including amplification temperature and amplification time were optimized. The optimized RT-LAMP can be done at 65 ℃ within 60 min. The detection limits of the RT-LAMP was 23.2 pg • µL-1. When the SYBR greenⅠdye was added to the amplification product，the positive samples were observed visually to be green，whereas the negative samples were orange，and there is no need for gel electrophoresis analysis. The eastablished RT-LAMP was used to detect CLBV from 120 field samples. The detection rate of different source samples was between 0 and 15%，the result of the RT-LAMP was in accord with RT-PCR method.

Key words: citrus, citrus leaf blotch virus, LAMP