Background:Circulating miRNAs have been identified as potential biomarkers for the early diagnosis and monitoring of cancers. However, limitations of polymerase chain reaction (PCR)-based methods are currently delaying the transition of miRNA research into clinical practice. These include labour-intensive workflows, exposure to errors and difficulties in detecting and quantifying low-abundance miRNAs.Objectives: This review emphasizes the need to develop amplification-free (“PCR-free”) technologies to improve the reliability, scalability and practicality of miRNA diagnostics in clinical settings.Methods:This review explores recent advances in PCR-free technologies developed over the past five years. It focuses on innovative methods, such as bead-based assays and sensor detection platforms, which serve as valuable alternatives to conventional PCR-based approaches. These emerging technologies have the potential to overcome the key limitations of PCR by offering streamlined workflows, reduced error rates and enhanced compatibility with a variety of clinical sample types. Crucially, they enable absolute quantification without the need for pre-nucleic acid extraction, reverse transcription or amplification, as well as the simultaneous detection of multiple miRNAs within a single assay. These provide cost-effective and scalable solutions for comprehensive biomarker profiling. The transition from PCR-based to PCR-free technologies is a significant step forward in miRNA diagnostics, overcoming long-standing technical barriers and paving the way for broader adoption of miRNA analysis in routine clinical settings. This shift supports the advancement of precision medicine and holds promises for improving early cancer detection.
背景：循环miRNA已被确定为癌症早期诊断和监测的潜在生物标志物。然而，基于聚合酶链反应（PCR）的方法存在局限性，目前阻碍了miRNA研究向临床实践的转化。这些局限性包括工作流程繁琐、易产生误差以及难以检测和定量低丰度miRNA。 目的：本综述强调需要开发无扩增（“无PCR”）技术，以提高miRNA诊断在临床环境中的可靠性、可扩展性和实用性。 方法：本综述探讨了过去五年中无PCR技术的最新进展。重点关注创新方法，如基于微珠的检测和传感器检测平台，这些方法可作为传统PCR方法的有价值替代方案。这些新兴技术通过提供简化的工作流程、降低错误率以及增强与各种临床样本类型的兼容性，有望克服PCR的关键局限性。至关重要的是，它们无需核酸提取、逆转录或扩增即可实现绝对定量，并能在单次检测中同时检测多种miRNA。这些技术为全面的生物标志物分析提供了经济高效且可扩展的解决方案。从基于PCR的技术向无PCR技术的转变是miRNA诊断领域的重要进步，克服了长期存在的技术障碍，为miRNA分析在常规临床环境中更广泛的应用铺平了道路。这一转变支持精准医学的发展，并有望改善癌症的早期检测。
肿瘤(癌症)患者之家