建立一种高度敏感和特异的方法来检测乳腺癌（BC）患者中循环PIK3CA H1047R突变，并研究PIK3CA H1047R突变的患病率与临床表现之间的关联。以等位基因特异性方式设计合适的阻断剂并针对基于 PCR 的 PIK3CA H1047R 突变鉴定进行了优化。所建立的技术在 196 名招募的 BC 患者的无细胞 DNA 样本中得到了验证。使用正确设计的阻断剂进行的等位基因特异性 PCR 测定能够检测到 0.01% 的 H1047R 突变体。通过应用新建立的检测方法，发现 62 例病例（占招募病例总数的 31.6%）携带血液循环 H1047R 突变体。其中，检测到的突变率随着疾病分期的增加而增加，从I期的2/18（11.1％）到II期的17/71（23.9％），III期的20/53（37.7％）和23/31（42.6％）。 %）的第四阶段（p = 0.025），分别。 H1047R 突变频率较高与晚期 (p = 0.033) 或复发 (p = 0.045) 或转移性患者 (p = 0.049) 以及接受放射治疗的人表皮生长因子受体 2 (HER2) 阳性 BC (p = 0.004）。 PIK3CA突变携带者常见于50岁以下肝转移、脑转移或淋巴结侵犯的患者中（p < 0.05）。成功建立了一种高灵敏度的新型等位基因特异性PCR检测方法，用于检测PIK3CA突变携带者。临床实践中的 PIK3CA H1047R 突变。版权所有：© 2024 Thao 等人。这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

To establish a highly sensitive and specific approach for the detection of circulating PIK3CA H1047R mutation in breast cancer (BC) patients and to investigate the association between the prevalence of PIK3CA H1047R mutation and clinical presentations.A proper blocker was designed in an allele-specific manner and optimized for PCR-based identification of the PIK3CA H1047R mutation. The established technique was validated in cell-free DNA samples from 196 recruited BC patients.The allele-specific PCR assay with a properly designed blocker was able to detect the H1047R mutant variant with 0.01%. By applying the newly established assay, 62 cases (31.6% of the total recruited cases) were found to carry a blood-circulating H1047R mutant. Wherein, the detected mutant rates increased with disease stages from 2/18 (11.1%) of stage I to 17/71 (23.9%) of stage II, 20/53 (37.7%) of stage III, and 23/31 (42.6%) of stage IV (p = 0.025), respectively. Higher frequencies of H1047R mutation were associated with late-stage (p = 0.033) or recurrence (p = 0.045) or metastatic patients (p = 0.049) as well as radiation-treated human epidermal growth factor receptor 2 (HER2) positive BC (p = 0.004). PIK3CA mutant carriers were frequently observed in patients under the age of 50 who had liver-metastasized or brain metastases or lymph node-invaded (p < 0.05).A novel allele-specific PCR assay with high sensitivity was established successfully for the detection of the PIK3CA H1047R mutation in clinical practice.Copyright: © 2024 Thao et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.