本研究為設計一嵌段高分子材料以偵測全血中的鼠疫桿菌(Yersinia pestis)，在檢測的同時屏除白血球的干擾。基材為柱/間距比率為1：1.5的透過翻模製備的圖案化聚二甲基矽氧烷(Poly(dimethylsiloxane), PDMS)表面接枝高分子，使之呈現柱型結構為500nm與1µm的一維光柵。利用可逆加成-斷裂鏈轉移聚合(Reversible Addition-Fragmentation Chain Transfer Polymerization, RAFT)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)的高分子，而後以點擊反應接枝於基材上。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，達到抗沾黏且可專一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，對不同的層數高分子製備嵌段高分子刷，測試出在四種樣品中PMDS-PSBMA-b-PMAA對Yersinia pestis專一性抓取的雷射能量變化最大為32.69%及43.88%，且在白細胞沾黏導致的雷射能量變化量最小為4.31%及2.68%，故經過交叉綜合考量，選其作為後續實驗之基材。
接著對於0 CFU/ml -10^6 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7.5×10^1 CFU/ml，雷射變化值大約為10%，而螢光顯微鏡已無法發現鼠疫桿菌的螢光存在，說明利用雷射能量檢測鼠疫桿菌的靈敏度較螢光顯微鏡高。而柱型500nm的圖案化PDMS對於不同濃度的鼠疫桿菌所作出的線性回歸，可以看到斜率較高為0.828代表有較高的靈敏度，且r2值為0.9738代表鼠疫桿菌濃度有97.38%的概率能以雷射解釋，非常有潛力作為即時檢測鼠疫桿菌的技術。

We successfully fabricated patterned PDMS sustrates with PMAA and PSBMA block copolymer brush which use RAFT then grafting to the surface by Click reaction in this experiment.

The results show that we have the polymer brush grafts to patterned PDMS sustrates in an aspect ratio of 1:1.5 which one dimension pillar structure grating width is 500nm and 1µm . Use AFM to investigate the height relationship with other factors 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.

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 optical microscope to analysis fluorescence intensity of modified patterned polymer brush combined with Yersinia pestis.

The block copolymer unit is prepared and graft to PDMS substrates. Polymer brushes tested that the sensitivity of PDMS-PSBMA-b-PMAA in the presence of the laser in human blood was superior to other substrates. It shows better sensitivity to Yersinia pestis and more than 99% anti-fouling ability of leukocytes.

The results showed that higher concentration of Yersinia pestis, lower intensity the laser diffraction we get. The sensitivity could reach a maximum concentration of 7.5×10^1 CFU/ml which laser energy loss is approximately 10%. However, the fluorescence of the Yersinia pestis could not be detected by optical microscope, indicating the sensitivity of the laser detection is higher than optical microscope fluorescent detection method.

The modificated PDMS with pillar width 500nm linear regression has the slope 0.828 meaning the better detection sensitivity for different concentrations of Yersinia pestis, and showed r2 0.9738. It will be considered a revolutionary tool for fast detection of Yersinia pestis.

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