Evans Ryan Thomas, Knudsen Katherine Elizabeth, Gillon-Zhang Elizabeth, Brown Julia Natalie, King Candace, Rossi Mary Beth, Kiser Cory, Schaffernoth James Alexander, Green Amanda Shull, Silva Ana-Luisa, von Bargen Kristine, Mordaka Justyna Malgorzata, Palmer Rebecca Natalie, Tomassini Alessandro, Collazos Alejandra, Andreazza Simonetta, Turner Iyelola, Ho Chau Ha, Nugent Dilyara, Jose Jinsy, Xyrafaki Christina, Barot Prarthna, Stolarek-Januszkiewicz Magdalena, Abujudeh Sam, Gray Eleanor Ruth, Gregg Jeffrey, Levin Wendy Jo, Balmforth Barnaby William, Pitts Kelly, Brown Shari
Biofidelity Inc., Morrisville, NC, USA.
Biofidelity Ltd., Cambridge, United Kingdom.
J Liq Biopsy. 2025 May 15;8:100298. doi: 10.1016/j.jlb.2025.100298. eCollection 2025 Jun.
Liquid biopsy is an important non-invasive method of sampling the molecular profile of tumors for patients to access personalized oncology therapeutics but can be challenging. NGS-based methods require high sample quality, high sequencing depth and associated cost, with complex workflows, while PCR assays are limited in variant coverage. Aspyre Clinical Test for Lung® (Blood) is a simplified genomic profiling assay for NSCLC that targets 114 variants in 11 genes () to robustly inform clinical management. The assay detects single nucleotide variants, insertions, deletions, gene fusions and exon skipping events from plasma-derived cfDNA and cfRNA simultaneously.
Sensitivity, specificity, analytical accuracy and analytical precision at standard input levels (20 ng cfDNA and 42 ng cfRNA) were tested using a combination of contrived samples and extracts from clinical samples taken from both healthy volunteers and patients with NSCLC. The effects of potential interfering substances on assay performance were tested. Assay sensitivity and specificity were also assessed at lower sample input levels (5 ng cfDNA and 6 ng cfRNA).
At standard input levels, median limits of detection were ≤0.25 % variant allele fraction for single nucleotide variants, ≤0.4 % variant allele fraction for insertions or deletions, ≤6 copies for gene fusions, and ≤100 copies exon 14 skipping events. The specificity from variant-free samples was 100 %. Tests of analytical accuracy yielded 100 % NPA and 94 % PPA between Aspyre Clinical Test for Lung (Blood) and either results from orthogonal NGS testing or expected outcomes of contrived samples. Results were 100 % replicable across multiple operators, reagent lots, days and equipment. At low input levels, median limits of detection were ≤0.8 % for single nucleotide variants and insertions/deletions, 6 copies for gene fusions and 100 copies for exon 14 skipping, with a false-positive rate of 0 %.
We present validation studies of Aspyre Clinical Test for Lung (Blood) using contrived and clinical samples. The technology is simple and fast, yet highly sensitive, specific, robust and reproducible with a turnaround time of two days. Aspyre Clinical Test for Lung (Blood) facilitates access to cost-effective, rapid, actionable molecular profiling of plasma for patients with NSCLC.
液体活检是一种重要的非侵入性方法，用于获取肿瘤患者的分子特征以进行个性化肿瘤治疗，但可能具有挑战性。基于二代测序（NGS）的方法需要高质量的样本、高测序深度及相关成本，且工作流程复杂，而聚合酶链反应（PCR）检测在变异覆盖方面存在局限性。肺癌Aspyre临床检测（血液）是一种针对非小细胞肺癌（NSCLC）的简化基因组分析检测方法，可检测11个基因中的114个变异（），以有力地指导临床管理。该检测可同时从血浆来源的游离DNA（cfDNA）和游离RNA（cfRNA）中检测单核苷酸变异、插入、缺失、基因融合和外显子跳跃事件。
使用人工合成样本和从健康志愿者及NSCLC患者采集的临床样本提取物的组合，在标准输入水平（20 ng cfDNA和42 ng cfRNA）下测试灵敏度、特异性、分析准确性和分析精密度。测试了潜在干扰物质对检测性能的影响。还在较低样本输入水平（5 ng cfDNA和6 ng cfRNA）下评估了检测的灵敏度和特异性。
在标准输入水平下，单核苷酸变异的检测限中位数≤0.25%变异等位基因分数，插入或缺失的检测限中位数≤0.4%变异等位基因分数，基因融合的检测限中位数≤6拷贝，外显子14跳跃事件的检测限中位数≤100拷贝。无变异样本的特异性为100%。肺癌Aspyre临床检测（血液）与正交NGS检测结果或人工合成样本的预期结果之间的分析准确性测试得出阴性预测值（NPA）为100%，阳性预测值（PPA）为94%。结果在多个操作人员、试剂批次、天数和设备之间的可重复性为100%。在低输入水平下，单核苷酸变异和插入/缺失的检测限中位数≤0.8%，基因融合的检测限中位数为6拷贝，外显子14跳跃的检测限中位数为100拷贝，假阳性率为0%。
我们展示了使用人工合成样本和临床样本对肺癌Aspyre临床检测（血液）进行的验证研究。该技术简单快速，同时具有高度的灵敏性、特异性、稳健性和可重复性，周转时间为两天。肺癌Aspyre临床检测（血液）有助于为NSCLC患者提供经济高效、快速且可指导治疗的血浆分子分析。
