Kadyrbayeva Rabiga, Kaidarova Dilyara, Shatkovskaya Oxana, Goncharova Tatyana, Orazgalieva Madina, Ossikbayeva Saniya
Chemotherapy Center, Kazakh Institute of Oncology and Radiology, Almaty, Republic of Kazakhstan -
Department of Oncology, Asfendiyarov Kazakh National Medical University, Almaty, Republic of Kazakhstan -
Panminerva Med. 2024 Dec;66(4):372-379. doi: 10.23736/S0031-0808.24.05172-3. Epub 2024 Sep 27.
The finding of mutations that activate epidermal growth factor receptor (EGFR) in people with lung adenocarcinoma resulted in the creation of a new class of biological treatments called tyrosine kinase inhibitors (TKI). These medications have changed how patients with EGFR mutations are clinically managed, nearly doubling their survival rate compared to standard chemotherapy. Though 1st and 2nd generation EGFR TKIs are initially highly effective, typically within 9-14 months all tumors with the mutation progress due to secondary resistance mutations involving alternative molecular pathways. In most cases (up to 60%), this is due to the T790M mutation emerging in the EGFR gene.
The study included 85 patients with NSCLC with progression of the disease after treatment with TKI 1st and 2nd generation. The T790M mutation was determined by digital polymerase chain reaction (PCR) on the QIAcuity One 5plex digital PCR system and traditional real-time PCR. Real-time PCR analysis of the presence of the T790M mutation was performed using the Therascreen EGFR Plasma RGQ PCR Kit (Qiagen). Using a digital PCR system in QIAcuity One (Qiagen) nanoplanets, the T790M mutation was analysed by digital PCR. The age of the patients ranged from 37 to 85 years.
Of 85 patients with NSCLC with disease progression after TKI treatment, T790M mutations were detected during digital PCR in 30 of 85 patients, which is 35.2% of the sample, and with traditional real-time PCR, positive mutations came out only in 3 out of 85 patients.
Thus, completed study can assert that digital PCR is able to replace traditional real-time PCR as a more preferable method of high-performance quantitative determination of target nucleic acids and has a relatively high sensitivity without compromising high specificity. Results of this research also show that a liquid biopsy using digital PCR provides an opportunity to avoid repeated tissue biopsy in patients who cannot provide a tumor tissue sample suitable for molecular analysis.
在肺腺癌患者中发现激活表皮生长因子受体（EGFR）的突变后，催生了一类名为酪氨酸激酶抑制剂（TKI）的新型生物治疗药物。这些药物改变了EGFR突变患者的临床管理方式，与标准化疗相比，其生存率几乎提高了一倍。尽管第一代和第二代EGFR TKI最初非常有效，但通常在9至14个月内，所有携带该突变的肿瘤都会因涉及替代分子途径的继发性耐药突变而进展。在大多数情况下（高达60%），这是由于EGFR基因中出现T790M突变。
该研究纳入了85例在接受第一代和第二代TKI治疗后疾病进展的非小细胞肺癌（NSCLC）患者。通过在QIAcuity One 5重数字PCR系统上进行数字聚合酶链反应（PCR）以及传统实时PCR来确定T790M突变。使用Therascreen EGFR Plasma RGQ PCR试剂盒（Qiagen）对T790M突变的存在进行实时PCR分析。在QIAcuity One（Qiagen）纳米孔板中使用数字PCR系统，通过数字PCR分析T790M突变。患者年龄在37至85岁之间。
在85例接受TKI治疗后疾病进展的NSCLC患者中，通过数字PCR在85例患者中的30例检测到T790M突变，占样本的35.2%，而使用传统实时PCR时，85例患者中仅3例检测到阳性突变。
因此，完成的研究可以断言，数字PCR能够替代传统实时PCR，作为一种更优的高性能定量测定靶核酸的方法，并且具有相对较高的灵敏度，同时不影响高特异性。本研究结果还表明，使用数字PCR的液体活检为无法提供适合分子分析的肿瘤组织样本的患者提供了避免重复组织活检的机会。
