本研究為設計一共聚高分子材料以偵測人血中的鼠疫桿菌(Yersinia pestis)，在檢測的同時屏除白細胞的干擾。基材為線／間距比率為1：1.5的圖案化光阻矽晶圓表面聚合高分子，使之呈現線寬為1、1.5、2、3μm的一維光柵。以此結構利用原子轉移自由基聚合(Atom Transfer Radical Polymerization, ATRP)製備甲基丙烯酸 (Poly(methacrylic acid)，PMAA)與甲基丙烯酸磺基甜菜鹼(Poly(sulfobetaine methacrylate)，SBMA)的共聚高分子刷。接著利用PMAA末端有-COOH官能基，透過EDC/NHS的反應接上Protein G與鼠疫桿菌F1抗原之單株抗體(Anti-Yp F1 MoAb)，進行鼠疫桿菌偵測，而PSBMA 的雙離子性可抵抗生物分子沾黏，達到抗沾黏且可抓取特定細胞的目的。
檢驗方式利用雷射因光柵繞射產生的能量散失，對不同的高分子比例製備共聚高分子刷，測試出在五種比例中PMAA:PSBMA(50:50)對Yersinia pestis專一性抓取的雷射能量變化量最大，且在白細胞沾黏導致的雷射能量變化量最小。
接著對於0 CFU/ml -106 CFU/ml做鼠疫桿菌檢體濃度靈敏度測試，並透過螢光顯微鏡觀察螢光佐證，結果顯示隨著鼠疫檢體濃度的上升，雷射繞射強度呈現下降的趨勢，其靈敏度可達到濃度101 CFU/ml，而螢光顯微鏡已無法發現鼠疫桿菌的螢光存在，說明雷射檢測鼠疫桿菌的靈敏度較螢光顯微鏡來的高。而線寬1um的圖案化晶片對於不同濃度的鼠疫桿菌有較佳的偵測靈敏度與定量能力，非常有潛力作為即時檢測鼠疫桿菌的技術。

We successfully fabricated patterned silicone wafer with PMAA and PSBMA co-polymer brush onto the surface by applying in-situ ATRP method in this experiment.

The results show that we have grown polymer brush in patterned silicon wafer in an aspect ratio of 1:1.5 which one dimension grating width is 1/ 1.5 / 2 / 3 um . By means of AFM to investigate the height relationship with other factor of polymer brush, XPS and FT-IR to analyze the constitution of the surface.

Subsequently, we use surface modification techniques with anti-body in order to capture Yersinia pestis by means of applying EDC/NHS reaction to combined carboxylic acid functional group of PMMA with Protein G and Anti-Yp (F1-MoAb). Fulfilling all the vacancy with BSA to change the surface with anti-fouling property, and then fully washed and test with Yersinia pestis detection.

The detection method theory utilizes laser energy loss due to grating diffraction. The sample concentration of 0 CFU/ml -106 CFU/ml is determined by using CLSM to analysis fluorescence intensity of modified patterned polymer brush combined with Yersinia pestis.

The copolymer unit is prepared and copolymerized at different polymer ratios. Polymer brushes tested that the sensitivity of PMAA:PSBMA (50:50) ratios in the presence of the laser in human blood was superior to other ratios performance which showed better sensitivity to Yersinia pestis and more than 99% anti-fouling ability of leukocyte.

The results showed that higher concentration of Yersinia pestis, lower intensity the laser diffraction we get. The sensitivity could reach a maximum concentration of 101 CFU/ml. However, the fluorescence of the Yersinia pestis could not be detected by CLSM, indicating the sensitivity of the laser detection is higher than CLSM fluorescent detection method.

The patterned wafer with a line width of 1um has better detection sensitivity for different concentrations of Yersinia pestis, and showed great potential as a tool for fast detection of Yersinia pestis.

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