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TAG-CLL是一种基于标记的新方法,用于对IGHV的体质突变检测,揭示了Sanger和下一代测序方法的弱点

Marzena Wojtaszewska¹, Monika Szelest², Marta Szarawarska³

¹Department of Hematology, University Clinical Hospital in Rzeszow, Rzeszow, Poland; Department of Hematology, College of Medical Sciences, University of Rzeszow, Rzeszow, Poland.

The Journal of molecular diagnostics : JMD

|September 25, 2025

概括

一个新的基于标记的工作流标准化了IGHV基因测序用于慢性淋巴细胞白血病 (CLL) 诊断. 这种方法可以在桑格和下一代测序平台之间进行直接比较,改善实验室一致性.

科学领域:

分子诊断学分子诊断
免疫遗传学免疫遗传学
在瘤学瘤学.

背景情况:

IGHV基因的体质突变 (SHM) 状态是慢性淋巴细胞白血病 (CLL) 的关键生物标志物.
目前的IGHV测序方法 (桑格和下一代测序) 在效率,成本和实验室间可比性方面存在挑战.
缺乏标准化的协议阻碍了诊断试验的可靠验证和基准测试.

研究的目的:

为IGHV基位引入一种基于标记的新型测序方法.
开发一个工作流程 (TAG-CLL),它与初始放大协议无关.
为了使IGHV测序结果在不同平台 (Sanger,IonTorrent,Illumina) 和实验室之间进行直接比较.

主要方法:

基于标记的图书馆准备方法被应用到来自人工橄克隆DNA样本的PCR安普利康.
十二个分子诊断实验室参与了这项研究,使用它们的标准协议来放大样本.
分类图书馆被排序,数据被分析为生产力,生殖系身份和SHM状态一致性.

主要成果:

TAG-CLL工作流程展示了在不同实验室协议和平台上比较IGHV测序的潜力.
分析揭示了放大系统偏差和识别的文物,为方法验证提供了洞察力.
在参与实验室中评估了SHM状态和生殖系身份的一致性.

结论:

本文提出的基于标记的方法为CLL诊断中的IGHV位点测序提供了一个标准化的框架.
这种方法促进了IGHV SHM测定和下一代测序免疫信息学管道的实验室间验证和基准测试.
工作流程解决了当前的方法挑战,为更可靠的CLL分子诊断铺平了道路.

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