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用于呼吸道合胞病毒检测的适体。

Aptamers for respiratory syncytial virus detection.

机构信息

Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary.

MTA-BME "Lendület" Chemical Nanosensors Research Group, Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, 1111, Budapest, Hungary.

出版信息

Sci Rep. 2017 Feb 21;7:42794. doi: 10.1038/srep42794.

Abstract

The identification of the infectious agents is pivotal for appropriate care of patients with viral diseases. Current viral diagnostics rely on selective detection of viral nucleic acid or protein components. In general, detection of proteins rather than nucleic acids is technically more suitable for rapid tests. However, protein-based virus identification methods depend on antibodies limiting the practical applicability of these approaches. Aptamers rival antibodies in target selectivity and binding affinity, and excel in terms of robustness and cost of synthesis. Although aptamers have been generated for virus identification in laboratory settings, their introduction into routine virus diagnostics has not been realized, yet. Here, we demonstrate that the rationally designed SELEX protocol can be applied on whole virus to select aptamers, which can potentially be applied for viral diagnostics. This approach does not require purified virus protein or complicated virus purification. The presented data also illustrate that corroborating the functionality of aptamers with various approaches is essential to pinpoint the most appropriate aptamer amongst the panel of candidates obtained by the selection. Our protocol yielded aptamers capable of detecting respiratory syncytial virus (RSV), an important pathogen causing severe disease especially in young infants, at clinically relevant concentrations in complex matrices.

摘要

鉴定病原体对于病毒性疾病患者的适当治疗至关重要。目前的病毒诊断依赖于对病毒核酸或蛋白质成分的选择性检测。一般来说,检测蛋白质而不是核酸在技术上更适合快速检测。然而,基于蛋白质的病毒识别方法依赖于抗体,这限制了这些方法的实际应用。适体在靶标选择性和结合亲和力方面可与抗体相媲美,并且在稳健性和合成成本方面表现出色。尽管已经在实验室环境中为病毒鉴定生成了适体,但它们尚未被引入常规病毒诊断中。在这里,我们证明了合理设计的 SELEX 方案可以应用于整个病毒以选择适体,这些适体有可能应用于病毒诊断。该方法不需要纯化的病毒蛋白或复杂的病毒纯化。所呈现的数据还表明,用各种方法验证适体的功能对于确定从选择中获得的候选适体面板中最合适的适体至关重要。我们的方案产生了能够检测呼吸道合胞病毒(RSV)的适体,RSV 是一种重要的病原体,尤其是在年幼的婴儿中,会导致严重的疾病,在复杂的基质中,能够以临床相关的浓度检测到 RSV。

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