Department of Materials, Department of Bioengineering and Institute for Biomedical Engineering, Imperial College London , London SW7 2AZ, U.K.
Department of Microbiology, Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev , Beer-Sheva 8410501, Israel.
ACS Nano. 2018 Jan 23;12(1):63-73. doi: 10.1021/acsnano.7b07021. Epub 2018 Jan 5.
Ebola virus disease causes widespread and highly fatal epidemics in human populations. Today, there is still great need for point-of-care tests for diagnosis, patient management and surveillance, both during and post outbreaks. We present a point-of-care test comprising an immunochromatographic strip and a smartphone reader, which detects and semiquantifies Ebola-specific antibodies in human survivors. We developed a Sudan virus glycoprotein monoplex platform and validated it using sera from 90 human survivors and 31 local noninfected controls. The performance of the glycoprotein monoplex was 100% sensitivity and 98% specificity compared to standard whole antigen enzyme-linked immunosorbent assay (ELISA), and it was validated with freshly collected patient samples in Uganda. Moreover, we constructed a multiplex test for simultaneous detection of antibodies against three recombinant Sudan virus proteins. A pilot study comprising 15 survivors and 5 noninfected controls demonstrated sensitivity and specificity of 100% compared to standard ELISA. Finally, we developed a second multiplex subtype assay for the identification of exposure to three related EVD species: Sudan virus, Bundibugyo virus and Ebola virus (formerly Zaire) using recombinant viral glycoprotein. This multiplex test could distinguish between the host's immunity to specific viral species and identify cross-reactive immunity. These developed serological platforms consisted of capture ligands with high specificity and sensitivity, in-house developed strips and a compatible smartphone application. These platforms enabled rapid and portable testing, data storage and sharing as well as geographical tagging of the tested individuals in Uganda. This platform holds great potential as a field tool for diagnosis, vaccine development, and therapeutic evaluation.
埃博拉病毒病在人群中引发广泛且致命的流行。如今,在爆发期间和之后,仍然非常需要用于诊断、患者管理和监测的即时检测。我们提出了一种即时检测方法,包括免疫层析条和智能手机读取器,用于检测和半定量人体幸存者中的埃博拉病毒特异性抗体。我们开发了苏丹病毒糖蛋白单克隆平台,并使用来自 90 名人类幸存者和 31 名当地未感染对照者的血清对其进行了验证。与标准全抗原酶联免疫吸附试验(ELISA)相比,糖蛋白单克隆的性能为 100%敏感性和 98%特异性,并在乌干达的新鲜采集的患者样本中进行了验证。此外,我们构建了一种用于同时检测三种重组苏丹病毒蛋白抗体的多重检测。一项包含 15 名幸存者和 5 名未感染对照者的试点研究表明,与标准 ELISA 相比,其敏感性和特异性为 100%。最后,我们开发了一种用于鉴定对三种相关埃博拉病毒物种(苏丹病毒、本迪布焦病毒和埃博拉病毒(前身为扎伊尔))暴露的第二种多重亚型检测方法,使用重组病毒糖蛋白。这种多重检测方法可以区分宿主对特定病毒物种的免疫和识别交叉反应性免疫。这些开发的血清学平台由具有高特异性和灵敏度的捕获配体、内部开发的条带和兼容的智能手机应用程序组成。这些平台能够实现快速和便携式检测、数据存储和共享以及对乌干达测试个体的地理标记。该平台作为一种用于诊断、疫苗开发和治疗评估的现场工具具有很大的潜力。