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为婴幼儿便的微RNA分析优化协议

David A Armstrong, Shannon M Soucy, Meghan E Muse

bioRxiv : the preprint server for biology

|April 16, 2025

概括

使用RNAlater或Zymo DNA/RNA Shield管与Zymo BIOMICs套件从婴儿便中获得了高质量的RNA和类似的microRNA档案. 这些发现支持使用婴儿便微RNA作为儿童发育和疾病的生物标志物的优化协议.

科学领域:

发现生物标志物的发现.
儿科研究的研究.
分子诊断学分子诊断学

背景情况:

微RNAs (miRNAs) 显示为儿童发育和疾病的生物标志物具有前景.
婴儿和幼儿便越来越多地用于转录基因分析.
挑战包括便样本中的高RNase水平和丰富的微生物RNA.

研究的目的:

为了比较婴幼儿便的RNA保存和提取协议.
评估商业套件和RNA质量和miRNA分析的保存方法.
在儿科便样本中建立miRNA分析的优化协议.

主要方法:

第一个阶段:使用碎片分析对RNA质量进行三种RNA提取套件和四种保存方法的比较.
第二阶段:利用miRNA测序 (miRNA-seq) 来比较在RNAlater中保存的便中的miRNA配置文件与Zymo DNA/RNA Shield管.
生物信息分析包括读数的分类 (人与微生物) 和对准miRBase v22.1.2.

主要成果:

齐莫BIOMICs套件产生了最高的RNA质量号码 (RQN).
RNAlater和Zymo DNA/RNA Shield管提供了可比较高质量的RNA产量.
两种保存方法之间没有观察到人类miRNA配置文件或检测到miRNA计数的显著差异.

结论:

在RNAlater或Zymo DNA/RNA Shield管中收集婴儿便,与Zymo BIOMICs套件相结合,产生高质量的RNA和一致的miRNA配置文件.
几种检测到的miRNAs (例如,miR-194a-3p,miR-200c-3p,miR-26a-5p) 都与肠道平衡有关.
这些发现为儿童便miRNA生物标志物研究的标准化方案提供了基础.

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