| Grapevine fleck disease caused by grapevine fleck virus (GFkV) is a worldly distributed disease. In China, the occurence of grapevine fleck disease was reported in Liaoning, Shangdong and Xinjiang. Usually the disease doesn't show any visible symptom on most grapevine cultivars and their rootstocks. Establishing a rapid method to detect GFkV is very an important role for the controllingof this virus disease.In this study, serological methods were employed to detected GFkV in 10 grapevine samples collected from the Fruit Tree Research Institute of Chinese Academy of Agricultural Science. The results showed that:there were four, two, one and two samples infected respectively by Grapevine Virus B (GVB), Grapevine FanLeaf Virus(GFLV), Grapevine Leaf Roll-associated Virus-1(GLRaV-1) and Grapevine Fleck Virus(GFkV). Protein A Sandwich Enzyme Link Immunosorbant Assay (PAS-ELISA), Plate Trapped Antigent-Indirec-ELISA (PTA-I-ELISA) and Triple Antibody Sandwich (TAS-ELISA) were respectively employed for the GFkV assay in grapevine phelome extraction solution in different dillution, the postive result was obtained when the crude of GFkV of the infected sample in the bark was diluted respectively by 1:200; 1:500; 1:2000. Both indirect Tissue Blot Immunoassay (DTBIA) and Dot-Immunobinding Assay (DIBA) were successfully applied to detect GFkV in the petiole and leaves, both have the advantages that the results were visual and less time was neededIn addition, Immunosorbant Electromicroscope (ISEM) was applied to GFkV detection in Shengbao sample, some spherical particles were founded, these particles were decorated by the polyclonal antibody of GFkV. These results indicated that the spherical particles were GFkVAn Immunocapture-ReverseTranscript-Polymerase Chain Reaction (IC-RT-PCR) was employed to detect GFkV in Shengbao, An about the 380bp fragement was amplified and it was longer than expected size... |
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