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解决基于microRNA的生物标志物开发中的未解决挑战:适合内源参考microRNA用于SARS-CoV-2感染严重程度

Thalia Belmonte¹, Manel Perez-Pons¹, Iván D Benítez¹

¹Translational Research in Respiratory Medicine, University Hospital Arnau de Vilanova and Santa Maria, IRBLleida, Lleida, Spain; CIBER of Respiratory Diseases (CIBERES), Institute of Health Carlos III, Madrid, Spain.

International journal of biological macromolecules

|April 30, 2024

概括

这项研究确定了miR-106a-5p和miR-484作为可靠的内源控制物,用于在住院COVID-19患者中正常化微RNA (miRNA) 定量PCR数据,确保准确的生物标志物开发.

科学领域:

生物化学生物化学
分子生物学分子生物学
传染性疾病传染性疾病

背景情况:

循环的无细胞微RNA (miRNAs) 显示出作为医学决策的生物标志物的潜力.
使用定量PCR (qPCR) 精确量化miRNAs需要稳定的内源参考基因进行正常化.
在SARS-CoV-2感染中,可复制miRNA生物标记的开发需要验证的参考miRNA.

研究的目的:

识别和验证稳定的内源基因miRNAs,用于QPCR数据正常化,用于SARS-CoV-2感染住院患者的血样本.
用多个算法和数据集评估候选参考miRNA的稳定性.

主要方法:

在住院COVID-19患者的170个血样本中使用RT-qPCR对179个miRNA进行分析.
使用geNorm,NormFinder和BestKeeper算法对候选miRNA的稳定性评估.
在关键的COVID-19患者样本中使用额外的RNA-seq数据集和RT-qPCR验证顶级候选miRNAs.

主要成果:

根据查阶段的稳定性,选择了11个候选miRNA.
所有算法都一致认为miR-106a-5p和miR-484是最高的内源控制.
选择的miRNAs表现出稳定的检测和低可变性,独立于临床或社会人口统计因素.

结论:

一个由两个miRNA小组 (miR-106a-5p和miR-484) 作为住院SARS-CoV-2患者的miRNA生物标志物研究的可靠正常化剂.
该小组将提高在COVID-19研究中基于qPCR的miRNA生物标志物开发的可复制性和准确性.
这些发现为推进循环miRNA在COVID-19临床决策中的使用提供了至关重要的工具.

关键词:

生物标志物生物标志物在 COVID-19 疫情中,内源性控制可以进行内源性控制.这是一个微型RNA.规范化规范化规范化参考基因是指基因中的基因.

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相关概念视频

MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...