Kehl Alexandra, Klein Ruth, Steiger Katja, Aupperle-Lellbach Heike
Laboklin GmbH&Co. KG, 97688 Bad Kissingen, Germany.
School of Medicine, Institute of Pathology, Technical University of Munich, 81675 München, Germany.
Vet Sci. 2025 Apr 21;12(4):390. doi: 10.3390/vetsci12040390.
Since microRNAs are released into the bloodstream and miRNA profiles are supposed to differ between healthy and tumour patients, miRNAs seem to have potential as biomarkers. An essential prerequisite for biomarkers in a routine diagnostic setup is their stability in serum over time. In this study, serum samples from 10 healthy dogs were analysed at different time points and under various temperature conditions (after 24 and 48 h, at 4° or 20 °C) for the copy number of eight miRNAs (miR-20b, 21, 122, 126, 192, 214, 222, 494) using ddPCR. The miR-21 had the highest copy number, whereas miR-494 had the lowest copy number in canine blood samples. The values of each miRNA varied individually between the dogs, showing a 5 to 10-fold range. Stability differed between the miRNAs, with miR-192 having the best stability. The copy number of miR-20b, miR-126 and miR-214 decreased not significantly during 48 h storage time. In contrast, miR-21, miR-122 and miR-222 were stable for 24 h only but decreased significantly after 48 h. The (in)stability of individual canine miRNAs must be considered when transferring study results into veterinary routine diagnostics, as the transport and storage conditions are variable. As far as possible, standardisation of sampling, storage and quantification of miRNAs is needed.
由于微小RNA会释放到血液中，而且健康患者和肿瘤患者的微小RNA谱理应不同，因此微小RNA似乎有作为生物标志物的潜力。在常规诊断设置中，生物标志物的一个基本前提条件是其在血清中随时间的稳定性。在本研究中，使用数字液滴聚合酶链反应（ddPCR）在不同时间点和各种温度条件下（24小时和48小时后，4°C或20°C）分析了10只健康犬的血清样本中8种微小RNA（miR-20b、21、122、126、192、214、222、494）的拷贝数。在犬类血液样本中，miR-21的拷贝数最高，而miR-494的拷贝数最低。每只犬的每种微小RNA的值各不相同，显示出5至10倍的范围。不同微小RNA的稳定性有所不同，其中miR-192的稳定性最佳。在48小时的储存时间内，miR-20b、miR-126和miR-214的拷贝数没有显著下降。相比之下，miR-21、miR-122和miR-222仅在24小时内稳定，但在48小时后显著下降。在将研究结果应用于兽医常规诊断时，必须考虑单个犬类微小RNA的（不）稳定性，因为运输和储存条件是可变的。尽可能需要对微小RNA的采样、储存和定量进行标准化。
