Caffrey Michael, Jayakumar Nitin, Caffrey Veronique, Anirudan Varada, Rong Lijun, Paprotny Igor
Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, IL 60607.
Department of Mechanical and Industrial Engineering, University of Illinois at Chicago, Chicago, IL 60607.
bioRxiv. 2024 Jan 3:2024.01.03.574055. doi: 10.1101/2024.01.03.574055.
The recent COVID-19 pandemic has underscored the danger of airborne viral pathogens. The lack of model systems to study airborne pathogens limits the understanding of airborne pathogen distribution, as well as potential surveillance and mitigation strategies. In this work, we develop a novel model system to study airborne pathogens using virus like particles (VLP). Specifically, we demonstrate the ability to aerosolize VLP and detect and quantify aerosolized VLP RNA by Reverse Transcription-Loop-Mediated Isothermal Amplification (RT-LAMP) in real-time fluorescent and colorimetric assays. Importantly, the VLP model presents many advantages for the study of airborne viral pathogens: (i) similarity in size and surface components; (ii) ease of generation and noninfectious nature enabling study of BSL3 and BSL4 viruses; (iii) facile characterization of aerosolization parameters; (iv) ability to adapt the system to other viral envelope proteins including those of newly discovered pathogens and mutant variants; (v) the ability to introduce viral sequences to develop nucleic acid amplification assays.
近期的新冠疫情凸显了空气传播病毒病原体的危险性。缺乏用于研究空气传播病原体的模型系统限制了我们对空气传播病原体分布的理解,以及潜在的监测和缓解策略。在这项工作中,我们开发了一种使用病毒样颗粒(VLP)来研究空气传播病原体的新型模型系统。具体而言,我们展示了将VLP雾化的能力,并通过实时荧光和比色测定中的逆转录环介导等温扩增(RT-LAMP)检测和定量雾化的VLP RNA。重要的是,VLP模型在研究空气传播病毒病原体方面具有许多优势:(i)大小和表面成分相似;(ii)易于产生且无感染性,能够研究BSL3和BSL4病毒;(iii)雾化参数易于表征;(iv)能够使系统适应其他病毒包膜蛋白,包括新发现病原体和突变变体的包膜蛋白;(v)能够引入病毒序列以开发核酸扩增检测方法。