Shen Kang, Hua Weiwei, Ge Shengjie, Mao Yu, Gu Yuexing, Chen Gaoyang, Wang Youwei
Department of Neurosurgery, The Affiliated Hospital of Yangzhou University, Yangzhou, China.
Institute of Translational Medicine, Medical College, Yangzhou University, Yangzhou, China.
Front Bioeng Biotechnol. 2023 Jan 13;10:1124441. doi: 10.3389/fbioe.2022.1124441. eCollection 2022.
Telomerase has been considered as a biomarker for early diagnosis and prognosis assessment of hepatocellular carcinoma (HCC), while the highly sensitive and specific methods remain challenging. To detect telomerase, a novel surface-enhanced Raman scattering (SERS) biosensor was constructed using the dual DNA-catalyzed amplification strategy composed of strand displacement amplification (SDA) and catalytic hairpin assembly (CHA). This strategy relies on the extension reaction of telomerase primer induced by telomerase, forming long-stranded DNAs with repetitive sequence to catalyze the follow-up SDA event. Subsequently, the SDA products can trigger the CHA reaction between the SERS probes (Au-Ag nanocages (Au-AgNCs) modified with hairpin DNA1 and Raman reporters) and capture substrate (Au@SiO array labeled with hairpin DNA2), resulting in the formation of numerous "hot spots" to significantly enhance the SERS signal. Results are promising that the established biosensor presented excellent reproducibility, specificity and sensitivity. Moreover, ELISA was applied as the golden standard to verify the application of the proposed biosensor in real samples and the results confirmed the satisfactory accuracy of our method. Therefore, the proposed SERS biosensor has the potential to be an ideal tool for the early screening of HCC.
端粒酶已被视为肝细胞癌（HCC）早期诊断和预后评估的生物标志物，然而，高灵敏度和高特异性的检测方法仍具有挑战性。为了检测端粒酶，采用由链置换扩增（SDA）和催化发夹组装（CHA）组成的双DNA催化扩增策略构建了一种新型表面增强拉曼散射（SERS）生物传感器。该策略依赖于端粒酶诱导的端粒酶引物延伸反应，形成具有重复序列的长链DNA，以催化后续的SDA事件。随后，SDA产物可触发SERS探针（用发夹DNA1修饰的金-银纳米笼（Au-AgNCs）和拉曼报告分子）与捕获底物（用发夹DNA2标记的Au@SiO阵列）之间的CHA反应，导致形成大量“热点”，从而显著增强SERS信号。结果表明，所建立的生物传感器具有优异的重现性、特异性和灵敏度。此外，将酶联免疫吸附测定（ELISA）作为金标准来验证所提出的生物传感器在实际样品中的应用，结果证实了我们方法的令人满意的准确性。因此，所提出的SERS生物传感器有潜力成为HCC早期筛查的理想工具。
