Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #06-01 Centros, Singapore, 138668, Singapore.
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Block MD7, 8 Medical Drive, Singapore, 117597, Singapore.
Sci Rep. 2017 Nov 30;7(1):16686. doi: 10.1038/s41598-017-16960-8.
The preservation of nucleic acids from clinical samples is critical to facilitate accurate molecular diagnosis. The use of a paper matrix, Flinders Technology Associates (FTA) Elute cards, to archive DNA and viral RNA is well-documented. However, the feasibility of FTA Elute cards for archiving serum and serum exosomal microRNAs (miRNAs) remains unclear. Here, we performed a comprehensive evaluation of FTA Elute cards for miRNA storage and recovery in different pre-analytical conditions. The recovery of serum miRNA dry-spotted on FTA Elute cards by direct elution with water at high temperature was poor. However, serum miRNAs dry-spotted on the cards were isolated with about 40% yield when using QIAzol lysis reagent and recovery was improved remarkably (>80%) upon extraction from cards pre-treated with trehalose. miRNAs stored on the cards remained stable at room temperature and can be kept for prolonged periods. Furthermore, miRNAs could be similarly recovered from serum exosomes dry-spotted on the cards. Importantly, when using sera from gastric cancer (GC) patients, the miRNAs were efficiently recovered from trehalose pre-treated cards without affecting their representation. Collectively, we have demonstrated the potential of FTA Elute cards to archive serum and serum exosomal miRNAs, making it useful for biomarker discovery and diagnostics.
从临床样本中保存核酸对于促进准确的分子诊断至关重要。使用纸质基质 Flinders Technology Associates (FTA) Elute 卡来存档 DNA 和病毒 RNA 的方法已有充分的文献记载。然而,FTA Elute 卡用于存档血清和血清外泌体 microRNAs (miRNAs) 的可行性尚不清楚。在这里,我们对 FTA Elute 卡在不同的预分析条件下用于 miRNA 储存和回收进行了全面评估。直接用高温水从 FTA Elute 卡上点样的血清 miRNA 回收效果很差。然而,当使用 QIAzol 裂解试剂时,从卡片上点样的血清 miRNAs 可以分离出约 40%的产量,并且在用海藻糖预处理的卡片上进行提取时回收率显著提高(>80%)。储存在卡片上的 miRNAs 在室温下稳定,可以长时间保存。此外,从点样在卡片上的血清外泌体中也可以类似地回收 miRNA。重要的是,当使用来自胃癌 (GC) 患者的血清时,从海藻糖预处理的卡片中可以有效地回收 miRNA,而不会影响其代表性。总的来说,我们已经证明了 FTA Elute 卡用于存档血清和血清外泌体 miRNA 的潜力,使其可用于生物标志物的发现和诊断。