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使用纳米抗体功能化伏安装置检测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)

SARS-CoV-2 detection using a nanobody-functionalized voltammetric device.

作者信息

Pagneux Quentin, Roussel Alain, Saada Hiba, Cambillau Christian, Amigues Béatrice, Delauzun Vincent, Engelmann Ilka, Alidjinou Enagnon Kazali, Ogiez Judith, Rolland Anne Sophie, Faure Emmanuel, Poissy Julien, Duhamel Alain, Boukherroub Rabah, Devos David, Szunerits Sabine

机构信息

Univ. Lille, CNRS, Centrale Lille, Univ. Polytechnique Hauts-de-France, UMR 8520 - IEMN, Lille, France.

Laboratoire d'Ingénierie des Systèmes Macromoléculaires (LISM), Institut de Microbiologie, Bioénergies et Biotechnologie (IM2B), Aix-Marseille Université - CNRS, UMR, Marseille, France.

出版信息

Commun Med (Lond). 2022 May 23;2:56. doi: 10.1038/s43856-022-00113-8. eCollection 2022.

Abstract

BACKGROUND

An ongoing need during the COVID-19 pandemic has been the requirement for accurate and efficient point-of-care testing platforms to distinguish infected from non-infected people, and to differentiate SARS-CoV-2 infections from other viruses. Electrochemical platforms can detect the virus via its envelope spike protein by recording changes in voltammetric signals between samples. However, this remains challenging due to the limited sensitivity of these sensing platforms.

METHODS

Here, we report on a nanobody-functionalized electrochemical platform for the rapid detection of whole SARS-CoV-2 viral particles in complex media such as saliva and nasopharyngeal swab samples. The sensor relies on the functionalization of gold electrode surface with highly-oriented Llama nanobodies specific to the spike protein receptor binding domain (RBD). The device provides results in 10 min of exposure to 200 µL of unprocessed samples with high specificity to SARS-CoV-2 viral particles in human saliva and nasopharyngeal swab samples.

RESULTS

The developed sensor could discriminate between different human coronavirus strains and other respiratory viruses, with 90% positive and 90% negative percentage agreement on 80 clinical samples, as compared to RT-qPCR.

CONCLUSIONS

We believe this diagnostic concept, also validated for RBD mutants and successfully tested on Delta variant samples, to be a powerful tool to detect patients' infection status, easily extendable to other viruses and capable of overcoming sensing-related mutation effects.

摘要

背景

在新冠疫情期间,一直需要准确高效的即时检测平台,以区分感染者和未感染者,并将严重急性呼吸综合征冠状病毒2(SARS-CoV-2)感染与其他病毒感染区分开来。电化学平台可以通过记录样品之间伏安信号的变化,通过病毒包膜刺突蛋白来检测病毒。然而,由于这些传感平台的灵敏度有限,这仍然具有挑战性。

方法

在此,我们报告了一种纳米抗体功能化的电化学平台,用于在唾液和鼻咽拭子样本等复杂介质中快速检测完整的SARS-CoV-2病毒颗粒。该传感器依赖于用对刺突蛋白受体结合域(RBD)具有高度定向性的羊驼纳米抗体对金电极表面进行功能化。该设备在接触200μL未处理样本10分钟后即可得出结果,对人类唾液和鼻咽拭子样本中的SARS-CoV-2病毒颗粒具有高度特异性。

结果

与逆转录定量聚合酶链反应(RT-qPCR)相比,所开发的传感器能够区分不同的人类冠状病毒株和其他呼吸道病毒,在80份临床样本上的阳性和阴性百分比一致性均为90%。

结论

我们认为这一诊断概念,也已在RBD突变体上得到验证,并在德尔塔变种样本上成功测试,是检测患者感染状态的有力工具,易于扩展到其他病毒,并且能够克服与传感相关的突变影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fd7/9126950/e990c307811a/43856_2022_113_Fig1_HTML.jpg

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