Several approaches for miRNA expression analysis have been developed in recent years. In this article, we provide an updated and comprehensive review of available qPCR-based methods for miRNA expression analysis and discuss their advantages and disadvantages. Existing techniques involve the use of stem-loop reverse transcriptase-PCR, polyadenylation of RNAs, ligation of adapters or RT with complex primers, using universal or miRNA-specific qPCR primers and/or probes. Many of these methods are oriented towards the expression analysis of mature miRNAs and few are designed for the study of pre-miRNAs and pri-miRNAs. We also discuss findings from articles that compare results from existing methods. Finally, we suggest key points for the improvement of available techniques and for the future development of additional methods.

中文翻译：

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基于qPCR的miRNA表达分析方法。

近年来，已经开发了几种用于miRNA表达分析的方法。在本文中，我们提供了用于miRNA表达分析的基于qPCR的可用方法的最新全面综述，并讨论了它们的优缺点。现有技术包括使用通用或miRNA特异性qPCR引物和/或探针，使用茎环逆转录酶PCR，RNA的聚腺苷酸化，衔接子或RT与复杂引物的连接。这些方法中有许多是针对成熟miRNA的表达分析而设计的，很少用于研究pre-miRNA和pri-miRNA。我们还将讨论比较现有方法结果的文章中的发现。最后，我们提出了一些关键点，以改进可用技术和未来开发其他方法。