Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, SE-752 37, Uppsala, Sweden.
Rarity Bioscience AB, SE-752 37, Uppsala, Sweden.
Nat Commun. 2022 Jul 12;13(1):4033. doi: 10.1038/s41467-022-31397-y.
Rare tumor-specific mutations in patient samples serve as excellent markers to monitor the course of malignant disease and responses to therapy in clinical routine, and improved assay techniques are needed for broad adoption. We describe herein a highly sensitive and selective molecule amplification technology - superRCA assays - for rapid and highly specific detection of DNA sequence variants present at very low frequencies in DNA samples. Using a standard flow cytometer we demonstrate precise, ultra-sensitive detection of single-nucleotide mutant sequences from malignant cells against up to a 100,000-fold excess of DNA from normal cells in either bone marrow or peripheral blood, to follow the course of patients treated for acute myeloid leukemia (AML). We also demonstrate that sequence variants located in a high-GC region may be sensitively detected, and we illustrate the potential of the technology for early detection of disease recurrence as a basis for prompt change of therapy.
在患者样本中罕见的肿瘤特异性突变可作为极好的标志物，用于在临床常规中监测恶性疾病的进程和治疗反应，并且需要改进检测技术以广泛采用。我们在此描述了一种高度敏感和选择性的分子扩增技术——超级 RCA 检测，用于快速、高度特异性地检测 DNA 样本中极低频率存在的 DNA 序列变异。我们使用标准流式细胞仪，针对骨髓或外周血中的正常细胞 DNA 高达 100,000 倍的过量，精确、超灵敏地检测来自恶性细胞的单核苷酸突变序列，以跟踪接受急性髓细胞性白血病 (AML) 治疗的患者的病程。我们还证明，位于高 GC 区域的序列变异可以被敏感地检测到，并说明了该技术用于早期检测疾病复发的潜力，作为及时改变治疗的基础。
