Sensitive, Low-Background-Signal miRNA Analysis via the Self-Priming-Initiated Color Reaction Loaded on a Rolling Circle Amplification Product,ACS Omega

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Sensitive, Low-Background-Signal miRNA Analysis via the Self-Priming-Initiated Color Reaction Loaded on a Rolling Circle Amplification Product
ACS Omega ( IF 3.7 ) Pub Date : 2024-01-26 , DOI: 10.1021/acsomega.3c06490
Meili Shao ₁ , Shaoli Wang ₁ , Bo Wang ₁ , Linqiong Guo ₁ , Yanping Cui ₁ , Yanling Wei ₂

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MicroRNAs (miRNAs) have been regarded as potential biomarkers in evaluating various diseases, such as pregnancy-induced hypertension and cancers. However, sensitive and reliable miRNA detection is still a challenge due to the low amplification efficiency and high background signal. Herein, we developed a colorimetric method for miRNA detection utilizing the self-priming-initiated color reaction loaded on a rolling circle amplification (RCA) product. In this method, a biotin-labeled RCA product is fixed on the surface of the streptavidin-coated wells, and the interfering components in samples are removed to avoid false reactions, thus reducing the background signals. Two signal amplification processes, including RCA and self-priming-initiated chain extension, endow the method with high sensitivity and a low limit of detection at the 10 fM level. In conclusion, our approach offers a promising perspective on sensitive and reliable miRNA detection and has the potential to be further utilized in biomedical research and early cancer detection.

中文翻译：

通过加载在滚环扩增产品上的自启动引发的颜色反应进行灵敏的低背景信号 miRNA 分析

MicroRNA (miRNA) 被认为是评估多种疾病的潜在生物标志物，例如妊娠高血压和癌症。然而，由于扩增效率低和背景信号高，灵敏可靠的 miRNA 检测仍然是一个挑战。在此，我们开发了一种用于 miRNA 检测的比色方法，利用加载在滚环扩增 (RCA) 产品上的自启动启动颜色反应。该方法将生物素标记的RCA产物固定在链霉亲和素包被的孔表面，去除样品中的干扰成分，避免错误反应，从而减少背景信号。包括 RCA 和自启动链延伸在内的两种信号放大过程使该方法具有高灵敏度和 10 fM 水平的低检测限。总之，我们的方法为灵敏可靠的 miRNA 检测提供了一个有前景的前景，并有可能进一步应用于生物医学研究和早期癌症检测。

更新日期：2024-01-26

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