本研究之核酸側向流免疫層析法,具備了檢測快速,且不具危險性, 在操作上安全性高。在膜上印製一級抗體(Rabbit anti Human IL-6)後,以三明治免疫反應捕捉抗原-人類白血球間質素-6(Interleukin 6),再利用有生物素(biotin)修飾的抗體(biotinylated Rat anti Human IL6),以鍊菌素(streptavidin)為橋梁連結一也有生物素修飾之環狀RCA環狀模板,再設計一段與其互補的環狀模板探針,結合後加入RCA所需材料直接在固相硝化纖維膜上進行恆溫DNA擴增,最後再以螢光探針顯色達到訊號放大。本研究針對低濃度的細胞激素-血白球間質素-6(IL-6)進行偵測,降低樣品之濃度,為主要目的。內文詳細敘述如何制備環狀模板與其在薄膜上偵測的方式,並了解在實驗中所遇到的各種問題與挑戰。
This study tilizes the DNA amplication technique on the Lateral-Flow nitrocellulose membrane to detect antigen Human IL-6。A sandwich immunoassay was built on the membrane,which was printed a primary antibody to capture the antigen(Human IL-6)。A RCA primer was designed with a sequence which could conjugate with the RCA circular template。The RCA circular template was then linked with the primary antibody by streptavidin。 The result was finally read out by the fluorescent signal。This study describes the preparation process ,the detect method, and problems encountered during the experiments。
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