Impact of ambient temperature exposure on miRNA stability in human plasma.

IF 2.5 4区工程技术 Q3 BIOCHEMICAL RESEARCH METHODS

BioTechniques Pub Date : 2025-07-01 Epub Date: 2025-09-08 DOI:10.1080/07366205.2025.2555657

Véronique Desgagné, Flore Lavoie, Imad Soukar, Marie-France Hivert, Luigi Bouchard, Perrie F O'Tierney-Ginn

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引用次数: 0

Abstract

MicroRNAs (miRNAs) are considered more stable than mRNA, but the impact of progressive thawing of biological samples after freezing as may happen during shipping delays has not been quantified. To address this, we utilized digital PCR to estimate the absolute concentrations of select miRNAs following progressive thawing of human plasma and maintenance at ambient temperature. Specifically, we quantified let-7b-3p, miR-144-5p, miR-150-5p, miR-517a-3p, miR-524-5p, and miR-1283, which have varying abundance in plasma. We observed a trend indicating a decline in miRNA concentration as plasma samples were progressively thawed. Notably, miR-150-5p and miR-517a-3p were the least stable and were degraded by 32% and 52% respectively after 24 hours of ambient temperature storage. We found that the variation in sensitivity to temperature was not due to the GC content of the miRNAs nor their initial abundance, suggesting that other factors, such as protein interactors and vesicles carrying these miRNAs, may impact sensitivity.

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环境温度暴露对人血浆中miRNA稳定性的影响。

MicroRNAs （miRNAs）被认为比mRNA更稳定，但由于运输延误，生物样品在冷冻后逐渐解冻的影响尚未被量化。为了解决这个问题，我们利用数字PCR来估计在人血浆逐渐解冻并在环境温度下维持后所选择的mirna的绝对浓度。具体来说，我们量化了血浆中丰度不同的let-7b-3p、miR-144-5p、miR-150-5p、miR-517a-3p、miR-524-5p和miR-1283。我们观察到，随着血浆样品逐渐解冻，miRNA浓度呈下降趋势。值得注意的是，miR-150-5p和miR-517a-3p是最不稳定的，在室温储存24小时后分别降解了32%和52%。我们发现对温度敏感性的变化不是由于mirna的GC含量或它们的初始丰度，这表明其他因素，如蛋白质相互作用物和携带这些mirna的囊泡，可能会影响敏感性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

BioTechniques 工程技术-生化研究方法

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3.3 months

期刊介绍： BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.