Triastuti Ayu, Zakiyyah Salma Nur, Gaffar Shabarni, Anshori Isa, Surawijaya Akhmadi, Hidayat Darmawan, Wiraswati Hesti Lina, Yusuf Muhammad, Hartati Yeni Wahyuni
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran Indonesia
Moleculer Biotechnology and Bioinformatics Research Center, Universitas Padjadjaran Indonesia
RSC Adv. 2023 Feb 16;13(9):5874-5884. doi: 10.1039/d2ra07560a. eCollection 2023 Feb 14.
A detection method based on an electrochemical aptasensor has been developed as an alternative fast, portable, simple, inexpensive, and high-accuracy detection method for detecting the SARS-CoV-2 Spike Receptor Binding Domain (spike RBD). The CeO@NH functionalized Screen Printed Carbon Electrode (SPCE) was used to immobilize an aminated aptamer of spike RBD protein glutaraldehyde as a linker. The aptamer's interaction with the SARS-CoV-2 Spike RBD was measured the [Fe(CN)] redox system signal. Experimental conditions were optimized using a Box-Behnken experimental design and showed that the optimal conditions of the SARS-CoV-2 aptasensor were 1.5 ng mL of aptamer, immobilization of aptamer for 60 minutes, and Spike RBD incubation for 10 minutes. The developed aptasensor was able to detect the standard SARS-CoV-2 Spike RBD with a detection limit of 0.017 ng mL in the range of 0.001-100 ng mL. This aptasensor was used to detect salivary and oropharyngeal swab samples of normal individuals with the addition of Spike RBD, and the recoveries were 92.96% and 96.52%, respectively. The testing on nasopharyngeal swab samples of COVID-19 patients showed that the aptasensor results were comparable with the qRT-PCR results. Thus, the developed aptasensor has the potential to be applied as a SARS-CoV-2 rapid test method for clinical samples.
一种基于电化学适体传感器的检测方法已被开发出来，作为一种替代方法，用于快速、便携、简单、廉价且高精度地检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突受体结合域（刺突RBD）。采用CeO@NH功能化丝网印刷碳电极（SPCE）固定刺突RBD蛋白的胺化适体，戊二醛作为连接剂。通过[Fe(CN)]氧化还原系统信号测量适体与SARS-CoV-2刺突RBD的相互作用。使用Box-Behnken实验设计优化实验条件，结果表明SARS-CoV-2适体传感器的最佳条件为适体浓度1.5 ng/mL、适体固定60分钟、刺突RBD孵育10分钟。所开发的适体传感器能够检测标准SARS-CoV-2刺突RBD，在0.001-100 ng/mL范围内检测限为0.017 ng/mL。该适体传感器用于检测添加了刺突RBD的正常个体的唾液和口咽拭子样本，回收率分别为92.96%和96.52%。对新冠肺炎患者鼻咽拭子样本的检测表明，适体传感器的结果与定量逆转录聚合酶链反应（qRT-PCR）结果相当。因此，所开发的适体传感器有潜力作为临床样本的SARS-CoV-2快速检测方法应用。
