金黃色葡萄球菌是造成嚴重感染和院內感染的重要病原菌，根據美國國家衛生研究院之院內感染監測系統統計，從1979到1995年院內感染病人身上培養出的菌株，金黃色葡萄球菌佔13%，且近年來有逐漸增加之趨勢，其快速鑑定愈形迫切。臨床檢驗上，免疫分析方法是目前最常用的檢驗技術，由於免疫分析方法具有相當高的特異性和靈敏度(1-10 ng/mL的抗原偵測量)，其應用歷久不衰。據報告顯示在1000個住院病人中得到菌血症的比例約為3.4至28人(平均10人)，死亡率約在15-30%。而引起菌血症的其中一種細菌正是金黃色葡萄球菌。現行免疫分析儀器缺點為售價高昂、測試項目過於窄化、操作煩瑣耗時且大多侷限於終點測試、部分並具有潛在危害（如放射性同位素）等。基於此，本研究對免疫分析技術想嘗試一革命性的改良，實驗步驟乃先製備直徑5 nm的奈米金(1×10-9 M)，以磷酸緩衝溶液配製不同濃度的抗體溶液，取得抗體與奈米金的最佳體積比例為10:1，純化後以OD280測定免疫金所接合的抗體含量是0.28，而達到最高接合抗體數目。並以點群C2v的對硫醇苯甲酸為分子探針，取濃度1×10-3 M的探針溶液20 l混合均勻即可，此分子在1073 cm-1和1580 cm-1具有很強的拉曼活性，再配合表面增顯拉曼散射將訊號放大10-100倍。最後，以金黃色葡萄球菌產生的特異性蛋白為模式系統，針對抗原濃度在10-5、10-6、10-7、10-8、10-9、10-10、10-11、10-12 g/ml做分析。結果顯示，迄今我們已可取得10-100 pg/ml的靈敏度與特異性，於傳統酵素免疫分析法比較可發現，其偵測極限高出1-2級數。此將可提供未來於單顆菌株之量測，因其可同時取得偵測目標生理結構的拉曼圖譜，故本免疫分析法，可同時得到定性與定量結果，不僅增加偵測極限，在未來應可再搭配介電泳晶片，發展成更快速、靈敏、簡易之方法，創立全新面貌之免疫分析系統。

Staphylococcus aureus is one of important pathogens, is always the orgin of infection in the hospital. Acroding to the statistics of the infectants, 13 percentages of the bacteria which detected was Staphylococcus aureus. There are trend increased year by year, and the rapid test is urgency. Immunoassays have been commonly used in the clinical laboratories for detection of a variety of antigens and antibodies. Basically, immunoassays are sensitive (around 1-10 ng/ml) and specific. The incident of bacteremia and fungemia has been reported to be 3.4-28/1,000 hospital admissions, and was estimated to an average of 10/1,000 (1%) around the world. The crude mortality rates of bacteremia ranged from 15 to 30%. The one of the common isolates from the blood cultures is staphylococcus aureus. The shortcoming of current immune analytical instrument is high price, narrow test project, convoluted operation and partly has potential to endanger (such as the radio isotope). We fabricated 5 nm (1×10-9 M) gold nanoparticles, and the antibody was diluted by PB (phosphate buffer). The optimal ratio of antibody to gold was 10:1, and the suspension of the antibody was 0.28 in A280. The point group of 4-mercapto- benzoic acid is C2v, and it can be used as molecular probe. The optimal concentration of mixed probe was 1×10-3 M. This probe has strong Raman signals in 1073 cm-1and 1585 cm-1, and can be enhanced to 1-2 order by gold nanoparticles. Protein A will be used as a model system, and its concentration was adjusted to be 10-5, 10-6, 10-7, 10-8, 10-9, 10-10, 10-11, 10-12 g/ml during the analysis. As a result, it showed that the detection of limit could reach to 10-100 pg/ml of antigen concentration and higher than the traditional method in sensitivity. The Raman scattering method may simultaneously provide the qualitative and quantitative analysis as a new immunoassay.

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