Advanced Medical and Dental Institute (AMDI), Universiti Sains Malaysia, Bertam, 13200, Kepala Batas, Penang, Malaysia; Department of Paraclinical Sciences, Faculty of Medicine and Health Sciences, Universiti Malaysia Sarawak, 94300, Kota Samarahan, Sarawak, Malaysia.
Institute of Health and Community Medicine, Universiti Malaysia Sarawak, 94300, Kota Samarahan, Sarawak, Malaysia.
Talanta. 2025 Jan 1;281:126842. doi: 10.1016/j.talanta.2024.126842. Epub 2024 Sep 13.
Chikungunya fever, caused by Chikungunya virus (CHIKV) exhibits clinical features that mimic that of other arbovirus infections such as dengue. CHIKV Envelope 2 (E2) protein, an antigenic epitope of CHIKV, has been identified as an ideal marker for diagnostics. The current CHIKV antigen detection tests are largely based on antibodies but are beleaguered by issues such as sensitivity to high temperature, expensive and prone to batch-to-batch variations. Aptamers are suitable alternatives to antibodies as they are cheaper and have no batch-to-batch variations compared to antibodies. In this study, DNA aptamer selection against CHIKV E2 proteins was performed using two different randomized ssDNA libraries. Chik-2 (96-mer) and Chik-3 (76-mer) were isolated from these two libraries and were identified as the potential aptamers against CHIKV E2 protein. The binding affinity of Chik-2 and Chik-3 against CHIKV E2 protein was estimated at 177.5 ± 32.69 nM and 30.01 ± 3.60 nM, respectively. A sandwich ELASA was developed, and this assay showed a detection limit of 2.17 x 10 PFU/mL. The sensitivity and specificity of the assay were 80 % and 100 %, respectively. The assay showed no cross-reactivity with dengue-positive samples, demonstrating the enormous diagnostic potential of these aptamers for the detection of CHIKV.
基孔肯雅热是由基孔肯雅病毒（CHIKV）引起的，其临床特征类似于登革热等其他虫媒病毒感染。CHIKV 包膜 2（E2）蛋白是 CHIKV 的一个抗原表位，已被确定为诊断的理想标志物。目前的 CHIKV 抗原检测试验主要基于抗体，但存在对高温敏感、昂贵且容易批次间变化等问题。与抗体相比，适体是抗体的合适替代品，因为它们更便宜，并且没有批次间变化。在这项研究中，使用两种不同的随机化 ssDNA 文库对 CHIKV E2 蛋白进行了 DNA 适体选择。从这两个文库中分离出 Chik-2（96 -mer）和 Chik-3（76-mer），并被鉴定为针对 CHIKV E2 蛋白的潜在适体。Chik-2 和 Chik-3 与 CHIKV E2 蛋白的结合亲和力分别估计为 177.5 ± 32.69 nM 和 30.01 ± 3.60 nM。建立了夹心 ELISA，该测定法的检测限为 2.17 x 10 PFU/mL。该测定法的灵敏度和特异性分别为 80%和 100%。该测定法与登革热阳性样本无交叉反应性，表明这些适体在检测 CHIKV 方面具有巨大的诊断潜力。
