The Impact of Tube Type, Centrifugation Conditions, and Hemolysis on Plasma Circulating MicroRNAs.

IF 3 3区医学 Q1 MEDICINE, GENERAL & INTERNAL

Diagnostics Pub Date : 2024-10-24 DOI:10.3390/diagnostics14212369

Belén Pastor-Navarro, Marta Ramírez-Calvo, Isabel Gil-Aldea, Isabel Cortell-Granero, José A López-Guerrero

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引用次数: 0

Abstract

Background: In recent years, liquid biopsy has emerged as a promising tool for the diagnosis and prognosis of numerous diseases, including cancer. Among the biomolecules analyzed in liquid biopsies are plasma circulating microRNAs (miRNAs), small non-coding RNAs that have proven to be crucial in the regulation of gene expression and the pathobiology of different health conditions, making them useful as biomarkers. However, variations in preanalytical conditions during biospecimen collection and processing can affect the analytical results.

Objectives: Herein, we determined how the type of collection tube, the number of centrifugations, and the degree of hemolysis can affect plasma circulating miRNA levels.

Methods: A cohort of 11 healthy donors was included. Whole blood was collected and handled in three different conditions, and miRNAs levels were analyzed using quantitative RT-PCR.

Results: Our results show that the differences in the type of preservative tubes influence hemolysis, measured as OD at 414 nm. Moreover, the number of centrifugations performed also altered miRNAs levels, increasing or decreasing them depending on the miRNA analyzed. Hence, our study shows that alterations in preanalytical conditions affect miRNAs levels, particularly the number of centrifugations and the type of collection tubes.

Conclusions: In our work, we highlight the importance of registering the preanalytical conditions in a standardized way that might be considered when analytical results are obtained.

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试管类型、离心条件和溶血对血浆循环 MicroRNA 的影响

背景：近年来，液体活检已成为诊断和预后包括癌症在内的多种疾病的有效工具。在液体活检分析的生物大分子中，有血浆循环微RNA（miRNA），这种小的非编码RNA已被证明在基因表达调控和不同健康状况的病理生物学中起着关键作用，因此可作为生物标记物。然而，生物样本采集和处理过程中分析前条件的变化会影响分析结果：在此，我们确定了采集管的类型、离心次数和溶血程度对血浆循环 miRNA 水平的影响：方法：纳入 11 名健康献血者。在三种不同条件下采集和处理全血，并使用定量 RT-PCR 分析 miRNA 水平：结果：我们的研究结果表明，防腐剂试管类型的不同会影响溶血（以 414 纳米波长的 OD 值测量）。此外，离心次数也会改变 miRNAs 的水平，根据所分析的 miRNA 的不同而有所增减。因此，我们的研究表明，分析前条件的改变会影响 miRNAs 的水平，尤其是离心次数和收集管的类型：在我们的工作中，我们强调了以标准化方式登记分析前条件的重要性，在获得分析结果时可能会考虑到这一点。

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

Diagnostics Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry

CiteScore

4.70

自引率

8.30%

发文量

2699

审稿时长

19.64 days

期刊介绍： Diagnostics (ISSN 2075-4418) is an international scholarly open access journal on medical diagnostics. It publishes original research articles, reviews, communications and short notes on the research and development of medical diagnostics. There is no restriction on the length of the papers. Our aim is to encourage scientists to publish their experimental and theoretical research in as much detail as possible. Full experimental and/or methodological details must be provided for research articles.