关键词: 柑橘, 柑橘黄化脉明病毒, 直接组织点免疫法

Abstract: Direct tissue blot immunoassay（DTBIA）was developed to detect Citrus yellow vein clearing virus（CYVCV）by exploring the optimal dilution of rabbit-anti-CYVCV polyclonal antibody （first antibody）and Alkaline phosphatase（Ap）conjugated goat-anti-rabbit IgG（secondary antibody）. The DTBIA was the most effective when the first antibody and secondary antibody diluted in 8 000 and 2 000 separately. Using the method，only CYVCV infected samples showed positive among detected samples associated with different citrus diseases；CYVCV could be effectively detected in 90 days after the tissue blotting membranes kept at 4 ℃. The consistency between DTBIA and one-step RT-PCR was 97.92% in CYVCV detection of citrus varieties. The results showed that the DTBIA method is rapid，simple，stable，specific and reliable，and can be applied for rapid detection of CYVCV in the field，which providing the important practical significance for control of the transmission of citrus yellow vein clearing virus and spreading of the disease widely.

Key words: Citrus, Citrus yellow vein clearing virus, direct tissue blot immunoassay