Selection of DNA aptamers against Chikungunya virus Envelope 2 Protein and their application in sandwich ELASA.

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2025-01-01 Epub Date: 2024-09-13 DOI:10.1016/j.talanta.2024.126842

Anna Andrew, Magdline S H Sum, Ewe Seng Ch'ng, Thean-Hock Tang, Marimuthu Citartan

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Abstract

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 10³ 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.

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针对基孔肯雅病毒包膜 2 蛋白的 DNA 嵌合体的筛选及其在夹心 ELASA 中的应用。

由基孔肯雅病毒（CHIKV）引起的基孔肯雅热的临床特征与登革热等其他虫媒病毒感染相似。CHIKV 包膜 2（E2）蛋白是 CHIKV 的抗原表位，已被确定为理想的诊断标记物。目前的 CHIKV 抗原检测测试主要以抗体为基础，但存在对高温敏感、价格昂贵、易出现批次间差异等问题。与抗体相比，适配体成本更低，而且不会出现批次间的差异，因此是抗体的合适替代品。本研究使用两种不同的随机 ssDNA 文库对针对 CHIKV E2 蛋白的 DNA 类似物进行了筛选。从这两个文库中分离出了 Chik-2（96-mer）和 Chik-3（76-mer），并确定它们是针对 CHIKV E2 蛋白的潜在适配体。据估计，Chik-2 和 Chik-3 与 CHIKV E2 蛋白的结合亲和力分别为 177.5 ± 32.69 nM 和 30.01 ± 3.60 nM。开发了一种夹心 ELASA，其检测限为 2.17 x 103 PFU/mL。检测灵敏度和特异性分别为 80 % 和 100 %。该测定与登革热阳性样本没有交叉反应，这表明这些灵敏剂在检测 CHIKV 方面具有巨大的诊断潜力。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.