乳腺癌是目前最廣泛的惡性腫瘤之一,可能發生在任何乳腺細胞中。乳癌也是全球女性最常見的癌症,因此對大範圍的人群造成重大影響。廣泛的研究顯示,Micro RNA對基因表現調控有重大的影響,它們在各種細胞過程中嚴格控制的表達,而Micro RNA失調則與癌症在內的許多病理狀況相關聯。在傳統的微滴式數位聚合酶鏈鎖反應(ddPCR)中,反應過程中的高溫熱循環會對微滴的油-水乳化殼層造成破壞,導致液滴破裂和融合,從而影響結果的準確性和可靠性。 相比之下,催化髮夾自組裝(Catalytic Hairpin Assembly, CHA)是一種不需酵素的等溫擴增技術,可以有效避免這些問題。CHA利用分子間的自組裝和相互作用,在等溫條件下實現目標分子訊號的高效擴增,從而避免了傳統ddPCR中因高溫循環導致的液滴不穩定性問題。這種方法不僅提高了檢測的準確性和可靠性,還簡化了操作流程,降低了實驗成本。 在本研究中,使用微流道系統結合催化髮夾自組裝技術,開發了一種名為droplet digital catalytic hairpin assembly(ddCHA)的方法來取代傳統的ddPCR。該研究利用微流道系統產生穩定的液滴,並在這些液滴內進行CHA反應,從而實現目標分子的高效檢測。這種創新方法不僅克服了ddPCR中存在的技術瓶頸,還為乳腺癌等疾病的早期診斷和監測提供了更加精確和可靠的工具。 ;Breast cancer is one of the most widespread malignant tumors and can occur in any breast cell type. As the most common cancer among women worldwide, it has a significant impact on global health. Extensive research shows that microRNAs play a crucial role in regulating gene expression and are associated with pathological conditions, including cancer, due to their aberration. Traditional droplet digital Polymerase Chain Reaction (ddPCR) faces challenges from high-temperature cycles, which destabilize droplet emulsions. In contrast, Catalytic Hairpin Assembly (CHA), an enzyme-free isothermal amplification method, avoids these issues, enhancing accuracy and simplifying workflows. This study introduces droplet digital catalytic hairpin assembly (ddCHA), combining CHA with microfluidic systems to create stable droplets for reactions. This innovative method overcomes ddPCR limitations and offers a precise tool for breast cancer diagnostics and monitoring.