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頁籤選單縮合
題名 | 水稻縞葉枯病毒偵測技術之發展=The Development of Immunological Detection Techniques of Rice Stripe Virus |
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作者 | 陳慶忠; 趙佳鴻; | 書刊名 | 臺中區農業改良場研究彙報 |
卷期 | 64 1999.09[民88.09] |
頁次 | 頁1-11 |
分類號 | 434.111 |
關鍵詞 | 水稻縞葉枯病毒; 媒介昆虫; 血清偵測技術; Rice stripe virus; Insect vector; Serologocal detection method; |
語文 | 中文(Chinese) |
中文摘要 | 應用酵素聯結法、組織轉漬法及西方浸漬法等技術偵測感染水稻縞葉枯病毒 (RSV) 之水稻組織及媒介昆蟲斑飛蝨之帶毒情形。利用夾層免疫酵素聯結法 (DAS ELISA)及間接免疫酵素聯結法,縞葉枯病毒抗血清稀釋至 32768倍,純化鞘蛋白及粗汁液抗原分別稀釋至10-5及10-6均能產生專一性免疫反應。以 DAS ELISA分析吸毒後且完成潛伏期之斑飛蝨帶毒蟲其 A405讀值介於 0.754~1.301間,未吸毒對照蟲則為 0.021~0.069。西方墨點法偵測吸毒後個別斑飛蝨,帶毒蟲在 RSV純化鞘蛋白 (34.7 K)相同位置產生專一性免疫反應。以組織轉漬法在硝化纖維膜擠壓罹病水稻根、莖、葉或斑飛蝨蟲體後,處理 RSV多元抗體及山羊白兔膽鹼酵素均能產生紫色反應,健康對照則無反應。試驗中斑飛蝨蟲體組織擠壓後會粘附於硝化纖維膜上而產生非專一性反應,造成對帶毒蟲個體判定之困擾。 |
英文摘要 | Rice stripe virus (RSV) disease has been recognized as the economically important disease on rice crop in Taiwan. In this study, some serological methods, including double-antibody sandwith (DAS) enzyme-linked immunosorbent assay (ELISA), indirect ELISA, tissue blotting on nitrocellulose membranes and western blotting were applied to detect RSV in RSV infected rice plant and its insect vector, Laodelphax striatellus. In DAS and indirect ELISA test, antiserum against RSV diluted at 32,768 fold showed positive reaction with purified coat protein and crude sap of RSV infected leaf at the dilution of 10-5 and 10-6, respectively. When using DAS ELISA to detect the individual viruliferous L. striatellus, the absorbance value at 405 nm were between 0.754 and 1.305, while those of the healthy insect were between 0.021 and 0.069. The western blot result showed that the viruliferous insect also reacted with antiserum. After direct tissue blotting and treated with polyclonal antibodies of RSV and goat-rabbit chondroitinase, the root, stem and leaf of infected rice plant or insect bodies all revealed the purple color. However, pressing the individual insect on the nitrocellulose membrane, it produced non-specific reaction, leading probably to a confusion for the detection. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。