在早期階段檢測類風濕性因子和抗環瓜氨酸肽抗體已被證明能有效預防和追縱類風濕性關節炎的發展以減少對關節的不可逆損害。然而，現有的方法往往只檢測單一目標，所以無法有效地提供準確的診斷信息。此外，它們是相對冗長（約90分鐘）、勞動密集型和昂貴的過程。因此，迫切需要一種新的方法來解決上述問題。在此，我們建立了一種在自動微流體平台上進行雙目標檢測的新方法。所開發的微流體晶片（45 mm x 62 mm）具有對稱設計的特點，可以在並行工作流程中同時進行雙目標檢測；它還具有自動化的優勢，可以減少人為錯誤。在這項研究中，通過使用表面塗有免疫球蛋白 G結晶片段和生物素化環瓜氨酸肽的磁珠，分別捕獲了兩種類風濕性關節炎的生物標誌物，類風濕性因子和抗環瓜氨酸肽抗體。在免疫磁性捕獲後，可經由測量螢光強度同時量化雙目標物的濃度。雙目標物（類風濕性因子和抗環瓜氨酸肽抗體）檢測能在 55 分鐘內以自動化方式完成，同時在消耗更少的樣品(5 µL)和試劑的條件下進行類風濕性關節炎的準確診斷。本研究說明了針對類風濕性關節炎中類風濕性因子和抗環瓜氨酸肽抗體的快速雙重檢測、更快、緊湊、靈敏的測量，為醫生提供有用和全面的診斷信息，以促進了類風濕性關節炎的準確診斷，從而改善了類風濕性關節炎的治療。

Detecting rheumatoid factor (RF) and cyclic citrullinated peptide antibodies (anti-CCP) at an early stage has been proven to be effective in preventing and tracking the development of rheumatoid arthritis (RA) and decreasing irreversible damage to joints. However, the existing methods often only detect a single target so accurate diagnostic information cannot be provided efficiently. Furthermore, they are relatively lengthy (about 90 min), labor-intensive, and expensive processes. A new approach to solve the above-mentioned issues is therefore of great need. Herein, a new approach of dual-target detection on an automated microfluidic platform was established. The developed microfluidic chip (45 mm x 62 mm) featured a symmetrical design that allows simultaneous dual-target detection with a parallel workflow; it also offers the advantage of automation to reduce human errors. In this study, two RA biomarkers, RF and anti-CCP, were respectively captured by using magnetic beads surface-coated with the fragment-crystallizable region of immunoglobulin G and biotinylated CCP. Following immunomagnetic capture, the fluorescence intensities were measured and the amounts of RF and anti-CCP were quantified simultaneously. Therefore, the dual-target (i.e. RF and anti-CCP) detection was carried out within 55 min and an accurate diagnosis of RA in an automatic format while consuming fewer samples (5 L) and reagents. The present work illustrates rapid dual-detection, faster, compact, and sensitive measurement of RF and anti-CCP in RA, providing physicians with useful and comprehensive diagnostic information to facilitate accurate RA diagnosis and thus improve RA treatment.

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