Chen Jiadong, Yu Qian, Lu Mengdan, Jeon Chang Su, Pyun Sung Hyun, Choo Jaebum
Department of Chemistry, Chung-Ang University, Seoul, 06974, South Korea.
R&D Center, Speclipse Inc., Seongnam, 13461, South Korea.
Anal Bioanal Chem. 2023 Oct;415(24):5939-5948. doi: 10.1007/s00216-023-04896-0. Epub 2023 Aug 17.
The development of rapid and accurate assays is crucial to prevent the rapid spread of highly contagious respiratory infections such as coronavirus (COVID-19). Here, we developed a surface-enhanced Raman scattering (SERS)-enzyme-linked immunosorbent assay (ELISA) method that allows for the screening of multiple patient samples with high sensitivity on a 1536-well plate. As the well number on the ELISA well plate increases from 96 to 1536, the throughput of the assay increases but the sensitivity decreases due to the low number of biomarkers and the increase in non-specific binding species. To address this problem, silica (SiO) beads were used to increase the surface-to-volume ratio and the loading density of biomarkers, thereby enhancing sensitivity. Using a three-dimensional gold nanoparticle (AuNP)@SiO SERS assay platform on a 1536-well plate, an immunoassay for the nucleocapsid protein biomarker of SARS-CoV-2 was performed and the limit of detection (LoD) decreased from 273 to 7.83 PFU/mL compared to using a two-dimensional assay platform with AuNPs. The proposed AuNPs@SiO SERS immunoassay (SERS-IA) platform is expected to dramatically decrease the false-negative diagnostic rate of the currently used lateral flow assay (LFA) or ELISA by enabling the positive diagnosis of patients with low virus concentrations.
开发快速准确的检测方法对于预防冠状病毒（COVID-19）等高传染性呼吸道感染的快速传播至关重要。在此，我们开发了一种表面增强拉曼散射（SERS）-酶联免疫吸附测定（ELISA）方法，该方法能够在1536孔板上对多个患者样本进行高灵敏度筛选。随着ELISA孔板的孔数从96增加到1536，检测通量增加，但由于生物标志物数量少和非特异性结合物种增加，灵敏度降低。为了解决这个问题，使用二氧化硅（SiO）珠来增加表面积与体积比以及生物标志物的负载密度，从而提高灵敏度。在1536孔板上使用三维金纳米颗粒（AuNP）@SiO SERS检测平台，对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的核衣壳蛋白生物标志物进行免疫测定，与使用二维AuNP检测平台相比，检测限（LoD）从273 PFU/mL降至7.83 PFU/mL。预计所提出的AuNPs@SiO SERS免疫测定（SERS-IA）平台能够通过对低病毒浓度患者进行阳性诊断，显著降低当前使用的侧向流动测定（LFA）或ELISA的假阴性诊断率。
