液滴数字 PCR 作为一种分子工具,用于检测具有表皮生长因子受体活化突变的 NSCLC 患者的表皮生长因子受体 T790M 突变。

IF 0.5 4区 医学 Q4 GENETICS & HEREDITY
Balkan Journal of Medical Genetics Pub Date : 2024-03-12 eCollection Date: 2023-12-01 DOI:10.2478/bjmg-2023-0020
S Durgut, L Salihefendić, D Pećar, I Čeko, N Mulahuseinović, M Izmirlija, R Konjhodžić
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引用次数: 0

摘要

背景:近50%的NSCLC患者最初对酪氨酸激酶抑制剂靶向疗法(TKI疗法)有成功的应答,但最终会出现获得性表皮生长因子受体T790M突变。T790M 二次突变可导致靶向治疗耐药和疾病复发。由于这种突变在液体活检样本中出现的频率很低,液滴数字 PCR(ddPCR)因其高灵敏度,为在 TKI 靶向治疗期间对疾病进行微创监测提供了可能:本研究共分析了 45 例 NSCLC 患者的血浆样本,这些患者都曾检测到表皮生长因子受体激活突变。提取的循环游离 DNA 经扩增后使用 ddPCR 技术检测是否存在 T790M 突变。数据分析使用 QuantaSoft 软件:结果:在检测的 45 份血浆样本中,共有 14 份样本被确定为 T790M 突变阳性。同样的样本最终在 FFPE 中也发现了 T790M 突变。液滴数字 PCR 在高灵敏度检测罕见等位基因变异方面显示出巨大优势。我们的 ddPCR 检测方法检测到的 T790M 突变等位基因的频率为 0.1%。ddPCR 产生的液滴平均数量为 9571 个:结论:监测 T790M 突变在检查处方 TKI 治疗效果方面具有重要作用。由于监测 TKI 治疗期间的潜在变化需要重复采样,我们的研究结果表明,ddPCR 技术使液体活检成为单核苷酸多态性 (SNP) 检测的一种适当的微创替代方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Droplet Digital PCR as a Molecular Tool for the Detection of the EGFR T790M Mutation in NSCLC Patients with the EGFR Activating Mutations.

Background: Almost 50% of NSCLC patients who initially show a successful response to tyrosine kinase inhibitors targeted therapy (TKI therapy) eventually develop acquired EGFR T790M mutation. The T790M secondary mutation can cause resistance to the targeted therapy and disease relapse. Since this mutation can be present at very low frequencies in liquid biopsy samples, droplet digital PCR (ddPCR), due to its high sensitivity, has opened the possibility for minimally invasive monitoring of the disease during TKI targeted therapy.

Materials and methods: For this study, a total of 45 plasma samples from NSCLC patients with previously detected EGFR-activating mutations were analyzed. Extracted circulating free DNA was amplified and examined for the presence of T790M mutation using ddPCR technology. For the data analysis, QuantaSoft Software was used.

Results: Of 45 tested plasma samples, a total of 14 samples were identified as positive for the T790M mutation. The same samples eventually showed the presence of T790M mutation in FFPE. Droplet digital PCR showed its great advantage in high sensitivity detection of rare allele variants. Our ddPCR assay detected T790M mutant allele in frequencies from 0.1%. The average number of droplets generated by ddPCR was 9571.

Conclusion: Monitoring of the T790M mutation has an important role in the examination of the effects of the prescribed TKI therapy. Since monitoring of potential changes during TKI therapy requires repeated sampling, our results showed that ddPCR technology has made it possible to use liquid biopsy as an adequate minimally invasive alternative for single nucleotide polymorphisms (SNP) detection.

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来源期刊
CiteScore
1.00
自引率
0.00%
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0
审稿时长
>12 weeks
期刊介绍: Balkan Journal of Medical Genetics is a journal in the English language for publication of articles involving all branches of medical genetics: human cytogenetics, molecular genetics, clinical genetics, immunogenetics, oncogenetics, pharmacogenetics, population genetics, genetic screening and diagnosis of monogenic and polygenic diseases, prenatal and preimplantation genetic diagnosis, genetic counselling, advances in treatment and prevention.
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