a Research Department , Centro Cardiologico Monzino IRCCS , Milan , Italy.
b Unit for the Study of Aortic, Valvular, and Coronary Pathologies , Centro Cardiologico Monzino IRCCS , Milan , Italy.
RNA Biol. 2018;15(10):1268-1272. doi: 10.1080/15476286.2018.1526538. Epub 2018 Oct 9.
Circulating microRNAs (miRNAs) are considered as reliable candidates for biomarker discovery. RNA-Sequencing has become the most suitable technique to accurately quantify the miRNAome. However, RNA-Sequencing relies on several technical passages before reaching the final-end. HTG EdgeSeq technology, thanks to the abrogation of RNA extraction step, allows productivity enhancement by reducing the number of hands-on steps, the time for sample preparation and, therefore, the costs. We found a strong correlation between qPCR and dPCR with HTG (Pearson's coefficient of 0.93 and 0.94, respectively). In conclusion, we showed that HTG EdgeSeq, performed on human plasma specimens without RNA extraction, is reliable, allows the simultaneous screening of more than 2,000 miRNAs, and thus, it could be applied to biomarker discovery in large cohorts.
循环 microRNAs(miRNAs)被认为是生物标志物发现的可靠候选者。RNA 测序已成为准确量化 miRNA 组的最适宜技术。然而,RNA 测序在到达最终结果之前需要经过几个技术步骤。HTG EdgeSeq 技术通过省去 RNA 提取步骤,减少了操作步骤的数量、样本制备的时间,从而降低了成本,提高了生产力。我们发现 HTG 与 qPCR 和 dPCR 之间具有很强的相关性(Pearson 系数分别为 0.93 和 0.94)。总之,我们表明,在没有 RNA 提取的情况下,在人血浆标本上进行 HTG EdgeSeq 是可靠的,允许同时筛选超过 2000 种 miRNAs,因此,它可以应用于大样本量的生物标志物发现。