以布朗運動檢測技術應用於齒舌蘭輪斑病毒檢測

Abstract

本研究成功開發一新式檢測方法，以布朗運動以及抗原與抗體間之專一性辨認做為檢測機制。實驗使用修飾上抗體之奈米粒子溶液與抗原齒舌蘭輪斑病毒(Odontoglossum ringspot virus, ORSV)溶液做反應，並以微粒子追蹤測速儀(micro-Particle Pracking Velocimetry, micro-PTV)進行量測與分析。結果顯示修飾上抗體的奈米粒子在加入抗原之後其布朗運動速度逐漸趨緩，且減緩趨勢隨抗原濃度增加而增大。 實驗亦確認了結果的重現性(Reproducibility)以及抗原與抗體間的辨識專一性(Specific Relationship)，且修飾過後的奈米粒子使用期限經實驗證實可達一年以上，除此之外，病株的實際檢測亦經確認可行。 本研究另以酵素連結免疫分析法(Enzyme-Linked Immunosorbent Assay, ELISA)之結果與本實驗做對照，除了驗證本實驗所用之抗體與抗原的辨認反應之外，亦提出本研究方法與酵素連結免疫分析法的比較及增加靈敏度的方式。此檢測方法能夠延伸應用於其他抗原與抗體間的辨識反應。

We have successfully developed a novel virus detection method based on antigen-antibody interactions and Brownian motion. The interactions between the antigen (Odontoglossum ringspot virus, ORSV) and nanobeads functionalized with the antibody (ORSV IgG) are recorded by micro-Particle Tracking Velocimetry (micro-PTV). The binding process in which ORSV conjugates to antibody-coated nanoparticles leads to an increase of the average diameter of the nanoparticles, the Brownian velocities therefore decrease. In other words, higher concentrations of ORSV will contribute to lower Brownian velocity of nanoparticles. Our experiments confirm the reproducibility and the specific relationship between the antibody and the antigen. The active lifespan of the functionalized nanoparticles, confirmed by the experiments, is at least one year. Besides, Virus detection in real condition is applicable as well. The results of Enzyme-Linked Immunosorbent Assay (ELISA) serve as the control groups and confirm the interactions between the antigen and the antibody. This economical and efficient method can also be optimized to reach the comparable reliability of ELISA under different conditions. Therefore, this method may be applied in further determination of other antigen-antibody interactions.