Center for Medical Genetics, Ghent University, Ghent, Belgium.
Novartis Institutes of Biomedical Research, Basel, Switzerland.
Nat Methods. 2014 Aug;11(8):809-15. doi: 10.1038/nmeth.3014. Epub 2014 Jun 29.
MicroRNAs are important negative regulators of protein-coding gene expression and have been studied intensively over the past years. Several measurement platforms have been developed to determine relative miRNA abundance in biological samples using different technologies such as small RNA sequencing, reverse transcription-quantitative PCR (RT-qPCR) and (microarray) hybridization. In this study, we systematically compared 12 commercially available platforms for analysis of microRNA expression. We measured an identical set of 20 standardized positive and negative control samples, including human universal reference RNA, human brain RNA and titrations thereof, human serum samples and synthetic spikes from microRNA family members with varying homology. We developed robust quality metrics to objectively assess platform performance in terms of reproducibility, sensitivity, accuracy, specificity and concordance of differential expression. The results indicate that each method has its strengths and weaknesses, which help to guide informed selection of a quantitative microRNA gene expression platform for particular study goals.
microRNAs 是蛋白质编码基因表达的重要负调控因子,在过去几年中得到了深入研究。已经开发了几种测量平台,以便使用不同的技术(例如小 RNA 测序、逆转录定量 PCR(RT-qPCR)和(微阵列)杂交)来确定生物样品中的相对 miRNA 丰度。在这项研究中,我们系统地比较了 12 种可用于分析 microRNA 表达的商业上可用的平台。我们测量了一组相同的 20 个标准化阳性和阴性对照样品,包括人类通用参考 RNA、人脑 RNA 及其稀释液、人血清样品和具有不同同源性的 microRNA 家族成员的合成 RNA 片段。我们开发了稳健的质量指标,以客观地评估平台在重现性、灵敏度、准确性、特异性和差异表达一致性方面的性能。结果表明,每种方法都有其优点和缺点,这有助于指导针对特定研究目标的定量 microRNA 基因表达平台的明智选择。