Backes Christina, Haas Jan, Leidinger Petra, Frese Karen, Großmann Thomas, Ruprecht Klemens, Meder Benjamin, Meese Eckart, Keller Andreas
Chair for Clinical Computational Biology, Saarland University, Saarbrücken, Germany.
Internal Medicine III, Heidelberg University, Heidelberg, Germany.
J Transl Med. 2015 Jul 14;13:224. doi: 10.1186/s12967-015-0594-x.
While in the past decades nucleic acid analysis has been predominantly carried out using quantitative low- and high-throughput approaches such as qRT-PCR and microarray technology, next-generation sequencing (NGS) with its single base resolution is now frequently applied in DNA and RNA testing. Especially for small non-coding RNAs such as microRNAs there is a need for analysis and visualization tools that facilitate interpretation of the results also for clinicians.
We developed miFRame, which supports the analysis of human small RNA NGS data. Our tool carries out different data analyses for known as well as predicted novel mature microRNAs from known precursors and presents the results in a well interpretable manner. Analyses include among others expression analysis of precursors and mature miRNAs, detection of novel precursors and detection of potential iso-microRNAs. Aggregation of results from different users moreover allows for evaluation whether remarkable results, such as novel mature miRNAs, are indeed specific for the respective experimental set-up or are frequently detected across a broad range of experiments.
We demonstrate the capabilities of miFRame, which is freely available at http://www.ccb.uni-saarland.de/miframe on two studies, circulating biomarker screening for Multiple Sclerosis (cohort includes clinically isolated syndrome, relapse remitting MS, matched controls) as well as Alzheimer Disease (cohort includes Alzheimer Disease, Mild Cognitive Impairment, matched controls). Here, our tool allowed for an improved biomarker discovery by identifying likely false positive marker candidates.
在过去几十年中,核酸分析主要使用定量的低通量和高通量方法,如定量逆转录聚合酶链反应(qRT-PCR)和微阵列技术,而现在具有单碱基分辨率的下一代测序(NGS)经常应用于DNA和RNA检测。特别是对于微小RNA等小非编码RNA,需要分析和可视化工具,以便临床医生也能理解检测结果。
我们开发了miFRame,它支持对人类小RNA NGS数据进行分析。我们的工具对已知前体中已知的以及预测的新型成熟微小RNA进行不同的数据分析,并以易于解释的方式呈现结果。分析包括前体和成熟微小RNA的表达分析、新型前体的检测以及潜在异微小RNA的检测等。此外,汇总不同用户的结果可以评估显著结果(如新型成熟微小RNA)是否确实特定于各自的实验设置,还是在广泛的实验中经常被检测到。
我们在两项研究中展示了miFRame的功能,该工具可在http://www.ccb.uni-saarland.de/miframe上免费获取,这两项研究分别是多发性硬化症的循环生物标志物筛查(队列包括临床孤立综合征、复发缓解型多发性硬化症、匹配对照)以及阿尔茨海默病(队列包括阿尔茨海默病、轻度认知障碍、匹配对照)。在这里,我们的工具通过识别可能的假阳性标志物候选物,改进了生物标志物的发现。