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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75997完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | 吳怡穎 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:17:06Z | - |
| dc.date.available | 2021-07-01T08:17:06Z | - |
| dc.date.issued | 1993 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75997 | - |
| dc.description.abstract | 為發展快速、簡易、經濟及適合養殖現場應用的診斷試劑，以調查臺灣養殖魚蝦貝類帶病原菌的情形，本研究首先以Vibrio vulnificus為對象。V.vulnificus為病原弧菌，不僅感染水生生物也會造成人類急性腸胃炎、傷口感染，甚至敗血症，影響公共衛生甚?。實驗首先選取可能為 V. vulnificus 的11個本地菌株和代表菌株 LSU 11-3M-30(biogroup l)在61℃(Tm-25℃)進行DNA雜交結合反應(DNA-DNA hybridization)，確定 E。8G、2-45c以及2-60b三菌株為V．vulnificus。再利用單株抗體P3C3，已知可專一辨識V. vulnificus 15個菌株的表面抗原，被覆(coating)於高密度(1.5g/cm3)乳膠微粒(latex beads)上，製成輔凝集反應試劑，結果顯示塗蓋抗體的最適濃度為500ug/ml，靈敏度可達106cells/ml。為瞭解乳膠凝集法之可行性，採集七次不同種的養殖生物，將樣本研磨、澄清、取上清液經12小時enrich等前處理後，僅需加人凝集試劑一個步驟約三十分鐘完成操作；此外，並以細菌分離鑑定法佐證乳膠凝集法的正確性及靈敏度。希望此診斷試劑的應用可推廣至養殖現場，加強業者對病害的監控，同時此試劑亦可應用於進出口海產品的檢疫及食品中毒等案例。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:17:06Z (GMT). No. of bitstreams: 0 Previous issue date: 1993 | en |
| dc.description.tableofcontents | 中文摘要 緒言------------------------------------------------------------------------------------------1 文獻回顧--------------------------------------------------------------------------------------3 材枓與方法------------------------------------------------------------------------------------7 一．細菌遺傳物質DNA之鑑定-------------------------------------------------------------------7 1．DNA之萃取------------------------------------------------------------------------------7 2．DNA-DNA雜交結合------------------------------------------------------------------------8 二．單株抗體的篩選與製備--------------------------------------------------------------------10 1．具專一性融合瘤細胞的篩選----------------------------------------------------------------10 2．腹水製備--------------------------------------------------------------------------------11 3．單株抗體的純化--------------------------------------------------------------------------12 三．乳膠凝集試劑的製備與測試-----------------------------------------------------------------12 1．單株抗體的被覆--------------------------------------------------------------------------12 2．抗原製備--------------------------------------------------------------------------------13 3．單株抗體濃度之測試----------------------------------------------------------------------14 4．乳膠凝集反應靈敏度之測試----------------------------------------------------------------14 5．乳膠凝集反應專一性之測試----------------------------------------------------------------14 四．樣品採集及分析---------------------------------------------------------------------------14 五．細菌鑑定---------------------------------------------------------------------------------15 1．溫度生長試驗----------------------------------------------------------------------------15 2．氨基酸去羧酸酵素之測定------------------------------------------------------------------16 3．纖維雙醣之利用--------------------------------------------------------------------------16 4．絲氨酸之利用----------------------------------------------------------------------------16 5．酪蛋白水解------------------------------------------------------------------------------16 6．酪氨酸之水解----------------------------------------------------------------------------17 7. ONPG測試--------------------------------------------------------------------------------17 8．弧菌抑制劑(O/129)試驗-------------------------------------------------------------------17 結果------------------------------------------------------------------------------------------18 一．DNA-DNA雜交結合反應---------------------------------------------------------------------18 二．對抗V.vulnificus單株抗體的篩選----------------------------------------------------------18 三．乳膠凝集試劑的測試----------------------------------------------------------------------19 1．被覆抗體濃度的選定---------------------------------------------------------------------19 2．專一性與靈敏度-------------------------------------------------------------------------19 四．台灣地區養殖鱸魚、石斑、鰻、吳郭魚、斑節蝦、牡蠣、文蛤帶V.vulnificus菌之調查------------19 討論------------------------------------------------------------------------------------------21 參考文獻--------------------------------------------------------------------------------------24 附圖------------------------------------------------------------------------------------------31 附表------------------------------------------------------------------------------------------33 附錄------------------------------------------------------------------------------------------42 | |
| dc.language.iso | zh-TW | |
| dc.title | 乳膠凝集法的開發：調查台灣養殖魚蝦貝類病原弧茵 Vibrio vulnificus | zh_TW |
| dc.title | Deve1opment of Latex agglutination test: Detection of Vibrio vu1nificus occurred in the cultured fish,shellfish and shrimp | en |
| dc.date.schoolyear | 81-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 48 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
| 顯示於系所單位： | 漁業科學研究所 | |
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