Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3631
標題: 染色介質poly-L-lysine搭配不同反應性染料對染料免疫分析的影響及多重待測物分析檢測的開發
Influence on Dye Immunoassay for Dyeing Poly-L-lysine with Different Reactive Dyes and the Development of Multi-analytes Assay Detection
作者: 李佳霖
Lee, Chia-Lin
關鍵字: immunoassay;免疫分析;reactive dye;poly-L-lysine;indirect labeling method;Schiff base;multi-analytes assay;反應性染料;聚離胺酸;間接標誌法;席夫鹼;多重待測物
出版社: 化學工程學系所
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摘要: 
本研究探討染料免疫分析中三種不同的反應性染料-Procion Turq. Bl、Levafix Blue CA和Taifix Red He-7BT對染色介質聚離胺酸(Poly-L-lysine, PLL)顯色強度的影響及多重待測物分析檢測的開發。利用間接標誌法並以人血清白蛋白(HSA)及其抗體(Anti-HSA)為模式檢體,改變席夫鹼(Schiff base)反應之pH值、染料濃度、反應時間和PLL染色溫度等四種實驗條件,來探討對試紙顯色強度的影響,可得到染料的較佳反應條件。再依據所得到的反應條件進行多株抗體HSA (Polyclonal anti-HSA)之競爭型分析檢測與開發在同一試紙上多重待測物分析檢測。
根據本研究所得的結果,可以得到以下結論:
1、三種反應性染料的較佳反應條件和HSA競爭型檢測範圍分別歸納如下:
(1) Procion Turq. Bl: pH值為6,染料濃度為5 .00 mg/ml,反應時間為90 鐘和染色溫度為35℃。競爭型檢測範圍為0 ~ 26.97 μg/ml,最低偵測濃度為0.19 μg/ml。
(2) Levafix Blue CA:pH值為8,染料濃度為 6.67 mg/ml,反應時間為60分鐘和染色溫度為35℃。競爭型檢測範圍為0 ~ 26.31 μg/ml,最低偵測濃度為1.25 μg/ml。
(3) Taifix Red He-7BT:pH 值為9,染料濃度為5.00 mg/ml,反應時間為30
分鐘和染色溫度為40℃。競爭型檢測範圍為0 ~ 17.78 μg/ml,最低偵測濃度為0.17 μg/ml。
2、標誌有不同染料之抗體Anti-HSA、Goat anti-rabbit IgG和Goat anti-swine IgG的混合液,能同步在同一張免疫層析試紙的測試線上分別和HSA、 Rabbit anti-VP2H和Anti-PRV特異性結合後,顯示不同顏色。

This research is to investigate the influence of color intensity on dyeing matrix poly-L-lysine for three different reactive dyes- Procion Turq. Bl, Levafix Blue CA and Taifix Red He-7BT and the development of the multi-analytes detection in dye immunoassay test. The reactive conditions may be influenced by using indirect labling method with Schiff base pH, dye concentration, reaction time and dyeing temperature to poly-L-lysine, respectively. The competitive assay of polyclonal anti-HSA to HSA can be done and the multi-analytes assay detection on the same strip may be developed on these reactive conditions
From the results of this study, the following results can be presented as:
1、The reactive conditions and the linear detection ranges of anti-HAS are listed as follows
(1) Procion Turq. Bl: pH 6, dye concentration: 5.00 mg/ml, reaction time: 90 mins and dyeing temperature: 35℃. The detection ranges of competitive assay: 0 ~ 26.97 μg/ml. The lower detection concentration: 0.19 μg/ml.
(2) Levafix Blue CA: pH 8, dye concentration: 6.67mg/ml, reaction time: 60 mins and dyeing
temperature: 35℃. The detection ranges of competitive assay: 0 ~ 26.31 μg/ml. The lower detection concentration: 1.25 μg/ml.
(3) Taifix Red He-7BT: pH 9, dye concentration: 5.00 mg/ml, reaction time: 30 mins and dyeing temperature: 40℃. The detection ranges of competitive assay: 0 ~ 17.78 μg/ml. The lower detection concentration: 0.17 μg/ml.
2、The different color can be shown on the testing line of the same strip after specific binding between anti-HSA, goat anti-rabbit IgG and goat anti-swine IgG in the mixing solution with HSA, rabbit anti-VP2H and anti-PRV, respectively.
URI: http://hdl.handle.net/11455/3631
其他識別: U0005-1308200716112200
Appears in Collections:化學工程學系所

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