文章：

从桑格测序到牛津纳米孔MinION技术：第三代测序对遗传性血液病诊断的影响

From Sanger to Oxford Nanopore MinION Technology: The Impact of Third-Generation Sequencing on Genetic Hematological Diagnosis

原文发布日期：29 May 2025

DOI: 10.3390/cancers17111811

类型: Article

开放获取: 是

英文摘要：

Background: Sanger sequencing remains the gold standard for characterizing genetic variants in short DNA fragments (<700 bp). However, the increasing demand for short TATs and high sensitivities in variant detection, particularly in oncohematology, is driving the need for more efficient methods. Next-generation sequencing (NGS) has improved sensitivity and allows for the simultaneous analysis of multiple genes, but it is still costly and time-consuming. Consequently, Sanger sequencing continues to be widely used. In this study, we have compared Sanger sequencing with Oxford Nanopore technology (ONT), which offers enhanced sensitivity and faster sequencing, delivering diagnostic results within 24 h. Methods: This study involves 164 samples (for a total of 174 analyzed regions of interest) previously characterized using either Sanger sequencing or a next-generation sequencing (NGS) panel, categorized by their genetic alterations. Validation was conducted on 15 genes crucial for the diagnosis, prognosis, or identification of drug resistance in myeloproliferative neoplasms (MPN), myelodysplastic syndromes (MDS), acute myeloid leukemia (AML), and chronic myeloid leukemia (CML). The primary objective was to assess whether MinION could identify the same variants previously detected in these patients. Results and Conclusions: With a 99.43% concordance observed in our comparison, our results support the implementation of MinION technology in routine variant detection in MPN, MDS, AML, and CML cases due to its significant advantages over Sanger sequencing.

摘要翻译： 

背景：桑格测序仍是短DNA片段（<700 bp）遗传变异表征的金标准。然而，对短周转时间（TAT）和高灵敏度变异检测的需求日益增长，特别是在血液肿瘤学领域，推动了对更高效方法的需求。下一代测序（NGS）提高了灵敏度并允许同时分析多个基因，但其成本仍然较高且耗时。因此，桑格测序仍被广泛使用。本研究比较了桑格测序与牛津纳米孔技术（ONT），后者具有更高的灵敏度和更快的测序速度，可在24小时内提供诊断结果。方法：本研究涉及164个样本（共174个分析的目标区域），这些样本先前已通过桑格测序或NGS panel进行了表征，并根据其遗传改变进行了分类。验证针对15个对骨髓增殖性肿瘤（MPN）、骨髓增生异常综合征（MDS）、急性髓系白血病（AML）和慢性髓系白血病（CML）的诊断、预后或耐药性识别至关重要的基因进行。主要目的是评估MinION是否能够识别这些患者先前检测到的相同变异。结果与结论：在我们的比较中观察到99.43%的一致性，我们的结果支持在MPN、MDS、AML和CML病例的常规变异检测中实施MinION技术，因为它相较于桑格测序具有显著优势。

原文链接：

From Sanger to Oxford Nanopore MinION Technology: The Impact of Third-Generation Sequencing on Genetic Hematological Diagnosis