Department of Neurosurgery, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People's Republic of China.
Department of Clinical Laboratory, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People's Republic of China.
Int J Nanomedicine. 2023 Jun 14;18:3211-3230. doi: 10.2147/IJN.S410080. eCollection 2023.
Circulating tumor DNA (ctDNA) is more representative and accurate than biopsy and is also conducive to dynamic monitoring, facilitating accurate diagnosis and prognosis of glioma. Therefore, the present study aimed to establish and validate a novel amplified method for the detection of IDH1 R132H and BRAF V600E, which were associated with the genetic diagnosis of glioma.
A dual-signal amplification method based on magnetic aggregation and catalytic hairpin assembly (CHA) was constructed for the simultaneous detection of ctDNAs. When target ctDNAs are present, the CHA reaction is initiated and leads to the assembly of Au-Ag nanoshuttles (Au-Ag NSs) onto magnetic beads (MBs). Further enrichment of MBs under an external magnetic field facilitated the dual-signal amplification of SERS.
The limit of detection (LOD) for IDH1 R132H and BRAF V600E in serum was as low as 6.01 aM and 5.48 aM. The reproducibility and selectivity of the proposed SERS analysis platform was satisfactory. Finally, the platform was applied to quantify IDH1 R132H and BRAF V600E in the serum of subcutaneous-tumor‑bearing nude mice, and the results obtained by SERS were consistent with those from quantitative real-time polymerase chain reaction (qRT-PCR).
The present study showed that the dual-signal amplification method is a simple and ultrasensitive strategy for gliomas-associated ctDNAs detection, which is crucial for early diagnosis and dynamic monitoring.
循环肿瘤 DNA（ctDNA）比活检更具代表性和准确性，也有利于动态监测，有助于准确诊断和预测胶质瘤。因此，本研究旨在建立和验证一种新的扩增方法，用于检测与胶质瘤遗传诊断相关的 IDH1 R132H 和 BRAF V600E。
构建了一种基于磁聚集和催化发夹组装（CHA）的双重信号扩增方法，用于同时检测 ctDNA。当存在靶 ctDNA 时，启动 CHA 反应，导致 Au-Ag 纳米梭（Au-Ag NSs）组装到磁性珠（MBs）上。在外磁场下进一步富集 MBs 促进了 SERS 的双重信号放大。
IDH1 R132H 和 BRAF V600E 在血清中的检测限（LOD）低至 6.01 aM 和 5.48 aM。所提出的 SERS 分析平台的重现性和选择性令人满意。最后，该平台被应用于定量皮下荷瘤裸鼠血清中的 IDH1 R132H 和 BRAF V600E，SERS 获得的结果与定量实时聚合酶链反应（qRT-PCR）一致。
本研究表明，双重信号扩增方法是一种用于检测胶质瘤相关 ctDNA 的简单和超灵敏策略，对于早期诊断和动态监测至关重要。
