0
78
（圖九）抗體間對表位競爭的示意圖
以 106 CFU/mL 經超音波破菌處理後的沙門氏菌進行 coating，並以我們生產的沙門氏菌 1 μg/mL 做為一級抗體進行偵測。
圖中(A)和(B)代表不同的兩種沙門氏菌單株抗體單獨偵測時所得之值，而(A+B)則為兩抗體混合進行偵測的值，(N)為以專 一性辨識大腸桿菌的單株抗體做為負對照組，(Non-competitive)表示兩抗體單獨偵測訊號的合(A)+(B)。當兩抗體並無表位 競爭性時(a)， (A+B) ≒ (A) + (B)；當兩抗體間有表位的競爭，或是兩者辨識的表位非常接近時(b、c)，則為 (A+B) ＜ (A) + (B)。
79
80
81
82
83
84
（圖十）沙門氏菌單株抗體間 competitive ELISA 實驗結果
依材料與方法（32 頁）中所述步驟進行 competitive EILSA。 (a)～(u)圖分別為 I-1～IV-3 七種抗體兩兩之間的 competitive ELISA 結果。圖中「MA-Eco」為以專一辨識大腸桿菌之單株抗體做為對照組；「Non-competitive」為兩抗體若無表位競 爭性的理論值。標示「**」者代表兩抗體間競爭訊號與無競爭之理論值間有顯著差異（p<0.05），且相差 5%以上；標示
「*」者代表兩抗體間競爭訊號與無競爭之理論值間雖有顯著差異，然而訊號相差在 5%以內。每個數據為三個重複樣品 的平均值為標準差。
0.0 0.4 0.8 1.2 1.6 2.0
OD
405-490**
(u)
85
86
87
（圖十一）沙門氏菌單株抗體對其抗原的作用力分析結果
以沙門氏菌（108 CFU/mL，經超音波破菌處理）做為抗原 coating 於 ELISA plate 中，以本研究中生產之沙門氏菌單株抗體稀釋至不同濃度（0.01 μg/mL 至 10 μg/mL）做為一級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行偵測。(a)
～(d)分別為第 I 群～第 IV 群抗體的 ELISA 實驗結果。每個數據為四個重覆樣 品的平均值及標準差。
88
（圖十二）I-1 抗體對破菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～106 CFU/mL，經超音波破菌處理）做為抗原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體（1 μg/mL）做為一級抗體，
10000 倍稀釋之 GAM-AP 為二級抗體，進行 direct ELISA 偵測。每個數據為四 個重複樣品的平均值及標準差。
0.0 0.2 0.4 0.6 0.8 1.0
1.E+03 1.E+04 1.E+05 1.E+06
OD
405-490破菌濃度 (CFU/mL)
I-1 MA-Eco
103 104 105 106
89 II-1 (boiled) MA-Eco (lived) MA-Eco (boiled)
103 104 105 106 107 108
90
（圖十三）II-1 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 II-1（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 II-1 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
91
III-1 (lived) III-1 (boiled) MA-Eco (lived) MA-Eco (boiled)
103 104 105 106 107 108
92
（圖十四）III-1 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 III-1（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 III-1 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
93
III-2 (lived) III-2 (boiled) MA-Eco (lived) MA-Eco (boiled)
103 104 105 106 107 108
94
（圖十五）III-2 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 III-2（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 III-2 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
95 MA-Eco (live) MA-Eco (boil)
103 104 105 106 107 108
96
（圖十六）IV-1 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 IV-1（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 IV-1 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
97 MA-Eco (live) MA-Eco (boil)
103 104 105 106 107 108
98
（圖十七）IV-2 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 IV-2（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 IV-2 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
99 IV-3 (boiled) MA-Eco (lived) MA-Eco (boiled)
103 104 105 106 107 108
100
（圖十八）IV-3 抗體對全菌抗原的 direct ELISA 分析結果
以不同濃度之沙門氏菌（103～108 CFU/mL，未經超音波處理之全菌）做為抗 原 coating 於 ELISA plate 中，再以沙門氏菌單株抗體 IV-3（1 μg/mL）做為一 級抗體，10000 倍稀釋之 GAM-AP 為二級抗體進行 direct ELISA 偵測。(a)圖 中，標示「lived」者表示 coating 之沙門氏菌未經煮沸處理，標示「boiled」者 表示 coating 之沙門氏菌經煮沸處理；(b)圖為截取(a)圖 IV-3 抗體辨識煮沸處理 過之沙門氏菌全菌（103～106 CFU/mL）之 direct ELISA 訊號。每個數據為四 個重複樣品的平均值及標準差。
101
（圖十九）Microfuge tube immunoassay 操作過程示意圖
102
（圖二十）IV-2 單株抗體對全菌抗原的 Microfuge tube immunoassay 結果 取經煮沸處理的沙門氏菌全菌樣品（103～106 CFU/mL）1.5 mL 於微量離心管 中，離心後去除上清液，加入10 μg/mL 沙門氏菌單株抗體 IV-2 昨為一級抗體、
5000 倍稀釋的 GAM-AP 做為二級抗體及 5000 倍稀釋的 RAG-AP 做為三級抗 體反應 2 小時，離心並去除上清液後加入受質呈色，最後取 50 μL 至 96 孔盤 於 ELISA reader 下讀取吸光值。圖中「MA-Eco」為加入專一辨識大腸桿菌之 單株抗體做為對照組。「*」標示指兩數據間有顯著差異（p<0.05）每個數據為
103
（圖二十一）本研究所使用的菌種分類表
104
（圖二十二）腸菌科中四種細菌（Klebsiella pneumoniae、Shigella sonnei、Escherichia coli 及 Salmonella enterica）之親 源性關係比較資料。本圖修繪自(Paradis et al., 2005)
105
（圖二十三）三種沙門氏菌菌株的 O 抗原種類及單體結構
本研究中進行菌株專一性分析時所選用的兩株 Typhimurium 血清型(a、b)及一株 Enteritidis 血清型(c) 的 O 抗原種類及單 體結構圖。「Abe」為 3, 6-dideoxy-D-galactose；「Tyv」3, 6-dideoxy-D-mannose。本圖修繪自(Wang et al., 2002)
106
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