Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR.
Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR.
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DOI：
10.1093/bib/bbv056
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发表时间：
2016-03
影响因子：
9.5
通讯作者：
Rossi RL
中科院分区：
文献类型：
--
作者：
Marabita F;de Candia P;Torri A;Tegnér J;Abrignani S;Rossi RL
健康和疾病个体血液中循环的microRNAs(MiRNAs)的高通量分析是生物标记物研究的活跃领域。虽然基于实时定量逆转录聚合酶链式反应(QPCR)的方法得到了广泛的应用，但数据应该如何归一化还没有解决。在这里，我们展示了不同算法的组合导致识别候选参考miRNAs，这些miRNAs可以在发现和验证阶段用作归一化因子。使用这里考虑的方法，我们确定了能够减少数据中非生物差异的归一化因子，并提供了几个案例研究，以说明在生理或病理场景的背景下的相关性。总而言之，从高通量研究中发现稳定的参考miRNAs使得聚焦的qPCR分析能够适当地标准化。
The high-throughput analysis of microRNAs (miRNAs) circulating within the blood of healthy and diseased individuals is an active area of biomarker research. Whereas quantitative real-time reverse transcription polymerase chain reaction (qPCR)-based methods are widely used, it is yet unresolved how the data should be normalized. Here, we show that a combination of different algorithms results in the identification of candidate reference miRNAs that can be exploited as normalizers, in both discovery and validation phases. Using the methodology considered here, we identify normalizers that are able to reduce nonbiological variation in the data and we present several case studies, to illustrate the relevance in the context of physiological or pathological scenarios. In conclusion, the discovery of stable reference miRNAs from high-throughput studies allows appropriate normalization of focused qPCR assays.