在遗传检测中对血液RNA测序的技术考虑:全球蛋白耗尽方法,批量效应和样本类型的评估
Xinyi Lu1, Yumeng Ma1, Xiaomei Luo1
1Department of Clinical Genetics Center, Shanghai Institute for Pediatric Research, Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Annals of laboratory medicine
|January 12, 2026
概括
基于试剂的球蛋白去除对于临床全血RNA测序是有效的,提供更好的基因检测,需要更少的读取. 周围血液单核细胞 (PBMC) 是基因诊断的首选样本类型.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 临床诊断 临床诊断 临床诊断
背景情况:
- 血液RNA测序 (RNA-seq) 对于诊断遗传疾病至关重要,但需要在临床使用中进行技术优化.
- 血液RNA-seq的关键技术因素包括全球蛋白耗尽,批量变异性和样本类型 (全血与外围血液单核细胞 [PBMCs]).
研究的目的:
- 为了评估基于试剂的全球蛋白去除 (globin-RR) 与生物信息全球蛋白去除 (globin-BR) 对于全血RNA-seq.
- 评估全球蛋白-RR样本中的批量效应,并比较全血与PBMCs的RNA-seq.
- 确定血液RNA-seq.的临床实施的最佳方法.
主要方法:
- 对效率和测序深度进行全球蛋白-RR和全球蛋白-BR的比较.
- 使用主要成分和相关性分析分析批量效应.
- 对比全血和PBMCs,以表达相关性和检测孟德尔病基因.
主要成果:
- 与全球蛋白-BR.R相比,Globin-RR显示出优越的全球蛋白消耗,更好地检测孟德尔病基因 (51.85%vs. 40.79%),并且需要较少的读数 (约1.07亿vs. ~1.49亿).
- 全球蛋白-RR显示了较低的批量效应 (批量间相关性,R2>0.96).
- 与全血相比,PBMC产生的孟德尔病基因 (3,675对3,598) 和高度表达的基因 (54.46%对51.85%) 的检测率更高,使它们成为首选的样本类型.
结论:
- 全球蛋白-RR是一种有效,可复制和实用的方法,用于临床全血RNA-seq.
- PBMCs提供高一致性和更广泛的基因检测范围,作为基因诊断测试的可行替代品.
- 优化的技术考虑提高了血液RNA-seq在遗传疾病诊断中的临床实用性.
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