Simple and Ultrasensitive Detection of Glioma-Related ctDNAs in Mice Serum by SERS-Based Catalytic Hairpin Assembly Signal Amplification Coupled with Magnetic Aggregation

Abstract

Youwei Wang,1,* Yang Yang,2,* Xiaowei Cao,3 Zhensheng Liu,4 Bing Chen,5 Qiu Du,1,* Xiaoxia Lu6,* 1Department of Neurosurgery, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China; 2Department of Clinical Laboratory, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China; 3Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China; 4Department of Interventional Radiology, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China; 5Department of neurosurgery, The Affiliated hospital of Qingdao University, Qingdao, Shandong Province, People’s Republic of China; 6Department of Oncology, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xiaoxia Lu, Department of oncology, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China, Tel [phone], Email [email protected] Qiu Du, Department of neurosurgery, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu Province, People’s Republic of China, Tel [phone], Email [email protected]: 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.Patients and Methods: 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.Results: 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).Conclusion: 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.Keywords: circulating tumor DNA, Au-Ag nanoshuttles, magnetic beads, catalytic hairpin assembly, surface-enhanced Raman spectroscopy

Keywords

• circulating tumor dna
• au-ag nanoshuttles
• magnetic beads
• catalytic hairpin assembly
• surface-enhanced raman spectroscopy