早期乳腺癌患者 ctDNA 检测中液滴式数字 PCR 与绝对 Q 数字 PCR 的对比研究。
Comparative study of droplet-digital PCR and absolute Q digital PCR for ctDNA detection in early-stage breast cancer patients.
机构信息
Unidad de Gestion Clinica Intercentros de Oncologia Medica, Hospitales Universitarios Regional y Virgen de la Victoria, 29010, Malaga, Spain; Centro de Investigacion Biomedica en Red de Cancer (CIBERONC - CB16/12/00481), 28029, Madrid, Spain.
Unidad de Gestion Clinica Intercentros de Oncologia Medica, Hospitales Universitarios Regional y Virgen de la Victoria, 29010, Malaga, Spain; The Biomedical Research Institute of Málaga (IBIMA-CIMES-UMA), 29010, Malaga, Spain.
出版信息
Clin Chim Acta. 2024 Jan 1;552:117673. doi: 10.1016/j.cca.2023.117673. Epub 2023 Nov 23.
BACKGROUND
Analysis of circulating tumor DNA (ctDNA) is increasingly used for clinical decision-making in oncology. However, ctDNA could represent ≤ 0.1 % of cell-free DNA in early-stage tumors and its detection requires high-sensitive techniques such as digital PCR (dPCR).
METHODS
In 46 samples from patients with early-stage breast cancer, we compared two leading dPCR assays for ctDNA analysis: QX200 droplet digital PCR (ddPCR) system from Bio-Rad which is the gold-standard in the field, and Absolute Q plate-based digital PCR (pdPCR) system from Thermo Fisher Scientific which has not been reported before. We analyzed 5 mL of baseline plasma samples prior to any treatment.
RESULTS
Both systems displayed a comparable sensitivity with no significant differences observed in mutant allele frequency. In fact, ddPCR and pdPCR possessed a concordance > 90 % in ctDNA positivity. Nevertheless, ddPCR exhibited higher variability and a longer workflow. Finally, we explored the association between ctDNA levels and clinicopathological features. Significantly higher ctDNA levels were present in patients with a Ki67 score > 20 % or with estrogen receptor-negative or triple-negative breast cancer subtypes.
CONCLUSION
Both ddPCR and pdPCR may constitute sensitive and reliable tools for ctDNA analysis with an adequate agreement in early-stage breast cancer patients.
背景
循环肿瘤 DNA（ctDNA）分析在肿瘤学领域越来越多地用于临床决策。然而，ctDNA 在早期肿瘤中的细胞游离 DNA 中可能仅占≤0.1%，其检测需要数字 PCR（dPCR）等高灵敏度技术。
方法
在 46 名早期乳腺癌患者的 46 份样本中，我们比较了两种用于 ctDNA 分析的领先 dPCR 检测方法：来自 Bio-Rad 的 QX200 液滴数字 PCR（ddPCR）系统，该系统是该领域的金标准，以及来自 Thermo Fisher Scientific 的 Absolute Q 板载数字 PCR（pdPCR）系统，该系统以前没有报道过。我们分析了基线治疗前 5 毫升的血浆样本。
结果
两种系统的灵敏度相当，突变等位基因频率无显著差异。事实上，ddPCR 和 pdPCR 在 ctDNA 阳性方面具有>90%的一致性。然而，ddPCR 表现出更高的可变性和更长的工作流程。最后，我们探讨了 ctDNA 水平与临床病理特征之间的关系。Ki67 评分>20%或雌激素受体阴性或三阴性乳腺癌亚型的患者 ctDNA 水平显著升高。
结论
ddPCR 和 pdPCR 均可作为早期乳腺癌患者 ctDNA 分析的敏感可靠工具，具有足够的一致性。
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