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題 名 | 比較酵素結合免疫吸附法與巢式聚合酶連鎖反應對山羊關節炎腦炎之檢測=Comparsion of ELISA and Nested PCR Assays for Detection of Caprine Arthritis Encephalitis |
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作 者 | 章嘉潔; 吳昇陽; | 書刊名 | 畜產研究 |
卷 期 | 49:1 2016.03[民105.03] |
頁 次 | 頁42-49 |
分類號 | 437.58 |
關鍵詞 | 山羊; 山羊關節炎腦炎; 酵素結合免疫吸附法; 巢式聚合酶連鎖反應; Goat; Caprine arthritis encephalitis; Enzyme linked immunosorbent assay; Nested PCR; |
語 文 | 中文(Chinese) |
中文摘要 | 本試驗旨在進行山羊關節炎腦炎 (Caprine arthritis encephalitis, CAE)血清監控及比較酵素結合免疫吸附法 (ELISA)與巢式聚合酶連鎖反應 (nested polymerase chain reaction, nested PCR)篩檢法,以提供控制本病方法之參考。試驗地點為臺東某羊場共 129頭努比亞山羊,分別採集受檢羊隻之頸靜脈血樣 10 mL,經離心取得血清後進行 CAE抗體檢測。本研究第一階段使用 ELISA,監測山羊 CAE狀況,血清學評估採 CAEV/MVV Antibody Test Kit (CHEKIT)商業套組,檢測結果所示 CAE檢出率 71.3% (92/129),在 3至 12、13至 24及 24月齡以上之不同年齡層, CAE檢出率分別為 77.9% (88/113)、54.5% (6/11)及 0% (0/5)。另羊隻依性別分類檢測 CAE結果所示,其中母羊檢測陽性反應檢出率為 72.6 % (45/ 62),公羊檢測陽性反應為 70.1% (47/ 67),結果顯示 CAE檢出率與性別無關。第二階段採血萃取 DNA,利用 nested PCR檢測山羊關節炎腦炎病毒 (Caprine arthritis encephalitis virus, CAEV)之 pol片段並定序確認,檢測 129頭羊呈陽性反應 97 (75.2%),增加 5頭陽性羊隻,進行卡伯統計 (Kappa statistic)分析結果 κ = 0.70,證實 ELISA和 nested PCR兩檢測結果呈現之高度一致性。 |
英文摘要 | The objectives of this experiment are to perform CAE (CAE) monitoring using competitive enzyme linked immunosorbent assay (ELISA) and nested polymerase chain reaction (nested PCR) assay Blood samples of 129 Nubian goats from a Taitung goat farm were used for test. CAE antibodies were detected in 71.3% (92/129) of serum samples using ELISA, In the results, the frequency of positive results in 3 to12 mons, 13 to 24 mons and above 24 mons were 77.9% (88/113), 54.5% (6/11) and 0% (0/5), respectively. The frequency of positive results in does and bucks were 72.6% (45/62) and 70.1% (47/67), respectively. There were no correlation between does and bucks in the CAE positive results (P > 0.05). DNA samples were extracted from the blood and used nested PCR assay targeting the CAEV proviral pol region, then nested PCR results were confirmed by sequence analysis, 97 (75.2%) goats were positive which increased the number of positive animals detected to 5, Kappa statistic showed substantial agreement between ELISA and nested PCR (κ = 0.70). |
本系統中英文摘要資訊取自各篇刊載內容。