Analytical and Clinical Validation of Solo-Test Driver: A Targeted Amplicon-Based NGS Test-System for FFPE and cfDNA Analysis in Clinical Oncology Setting

IF 2.6 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Clinical Laboratory Analysis Pub Date : 2025-03-08 DOI:10.1002/jcla.70008

Maxim Ivanov, Alexandra Lebedeva, Ekaterina Belova, Tatiana Grigoreva, Egor Veselovsky, Alexandra Kavun, Anastasiia Taraskina, Olesya Kuznetsova, Vladislav Nikulin, Laima Belyaeva, Daria Kravchuk, Tatyana Lisitsa, Alexey Barinov, Natalia Pospekhova, Saida Aliyarova, Ekaterina Khomenko, Alexey Tryakin, Irina Demidova, Anna Stroganova, Mikhail Fedyanin, Vladislav Mileyko

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引用次数: 0

Abstract

Background

Next-generation sequencing (NGS) is increasingly integrated into cancer patient management, necessitating cost-effective, reliable tests for companion diagnostics. We present the validation of the Solo-test Driver panel, a custom NGS amplicon-based tool for DNA analysis of 34 oncogenes, addressing key clinical needs.

Methods

The panel's performance was validated for detecting SNVs, CNVs, and MSI. Analytical validation used 182 samples, while clinical validation involved 130 samples, both encompassing diverse tumor types and specimen formats.

Results

The Solo-test Driver panel has the potential to identify an additional 18.3%, 16.5%, and 8.7% of RAS+ colorectal, PIK3CA+ breast, and EGFR+ lung cancer patients, respectively, when compared to FDA-approved PCR tests. Analytical validation demonstrated high intra-lab robustness (coefficient of variation of coverage uniformity 6.4%) and high inter-lab robustness (PCC of per-amplicon coverage 0.82, 0.84, 0.9 for three different labs). Estimated in silico sensitivity was 100% for detecting clinically actionable SNVs at 250x, corresponding to only 60,000 read pairs per sample. In vitro mixing experiments determined LoD starting from 3.3% VAF. Estimated in silico LoD ranged from 0.5% to 5% at 1000× (1% to 5% at 650x). Clinical validation demonstrated PPA/NPA of 100%/80%, 95%/100%, and 100%/100% for detecting MSI, SNVs, and CNVs, respectively.

Conclusions

The Solo-test Driver panel offers a reliable, cost-effective solution for detecting somatic alterations and genomic signatures, making it suitable for routine companion diagnostics in solid tumors.

Abstract Image

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单个测试驱动程序的分析和临床验证：一种基于靶向扩增子的NGS测试系统，用于临床肿瘤环境中的FFPE和cfDNA分析。

背景：下一代测序（NGS）越来越多地集成到癌症患者管理中，需要具有成本效益，可靠的伴随诊断检测。我们展示了Solo-test Driver面板的验证，这是一种基于NGS扩增子的定制工具，用于34种癌基因的DNA分析，解决了关键的临床需求。方法：验证该面板检测SNVs、CNVs和MSI的性能。分析验证使用了182个样本，而临床验证涉及130个样本，包括不同的肿瘤类型和标本格式。结果：与fda批准的PCR检测相比，单独检测驱动组有可能分别识别出18.3%、16.5%和8.7%的RAS+结直肠癌、PIK3CA+乳腺癌和EGFR+肺癌患者。分析验证表明，该方法具有较高的实验室内稳健性（覆盖均匀度变异系数为6.4%）和实验室间稳健性（每个扩增子覆盖的PCC在三个不同实验室分别为0.82、0.84和0.9）。估计在250倍时检测临床可操作的snv的硅灵敏度为100%，对应于每个样本只有60,000对读对。体外混合实验从3.3% VAF开始测定LoD。在1000倍的情况下，硅片的LoD估计为0.5%至5%（在650倍的情况下为1%至5%）。临床验证表明，PPA/NPA分别为100%/80%、95%/100%和100%/100%用于检测MSI、snv和cnv。结论：Solo-test Driver面板为检测体细胞改变和基因组特征提供了一种可靠、经济的解决方案，使其适用于实体肿瘤的常规伴随诊断。

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来源期刊

Journal of Clinical Laboratory Analysis 医学-医学实验技术

CiteScore

5.60

自引率

7.40%

发文量

584

审稿时长

6-12 weeks

期刊介绍： Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.