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磁性纳米捕获颗粒可保持委内瑞拉马脑炎病毒在血液中的稳定性以用于实验室检测。

Magnetic Nanotrap Particles Preserve the Stability of Venezuelan Equine Encephalitis Virus in Blood for Laboratory Detection.

作者信息

Akhrymuk Ivan, Lin Shih-Chao, Sun Mei, Patnaik Anurag, Lehman Caitlin, Altamura Louis, Minogue Timothy, Lepene Ben, van Hoek Monique L, Kehn-Hall Kylene

机构信息

National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA, United States.

United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD, United States.

出版信息

Front Vet Sci. 2020 Jan 28;6:509. doi: 10.3389/fvets.2019.00509. eCollection 2019.

Abstract

Most of the modern techniques used for identification of viral-induced disease are based on identification of viral antigens and/or nucleic acids in patient's blood. Diagnosis in the field or in remote locations can be challenging and alternatively samples are shipped to diagnostic labs for testing. Shipments must occur under controlled temperature conditions to prevent loss of sample integrity. We have tested the ability of magnetic Nanotrap® (NT) particles to improve stability and detection of Venezuelan equine encephalitis virus (VEEV), viral capsid protein, and viral genomic RNA in whole human blood at elevated temperature and prolonged storage conditions. NT particles have previously been shown to capture and enrich multiple pathogens including respiratory syncytial virus, influenza virus, coronavirus, and Rift Valley fever virus. Our study indicates that samples incubated with NT particles had detectable levels of infectious VEEV in blood equal to or greater than samples without NT treatment across all temperatures. Viral RNA detection was increased in the presence of NT particles at later time points (72 h) and higher temperature (40°C) conditions. Likewise, detection of VEEV capsid protein was enhanced in the presence of NT particles up to 72 h at 40°C. Finally, we intranasally infected C3H mice with TC-83, the live attenuated vaccine strain of VEEV, and demonstrated that NT particles could substantially increase the detection of VEEV capsid in infected blood incubated up to 72 h at 40°C. Samples without NT particles had undetectable capsid protein levels. Taken together, our data demonstrate the ability of NT particles to preserve and enable detection of VEEV in human and mouse blood samples over time and at elevated temperatures.

摘要

大多数用于识别病毒引起疾病的现代技术都是基于对患者血液中病毒抗原和/或核酸的识别。在现场或偏远地区进行诊断可能具有挑战性,因此样本通常会被送往诊断实验室进行检测。样本运输必须在可控的温度条件下进行,以防止样本完整性受损。我们测试了磁性纳米捕集器(NT)颗粒在高温和长时间储存条件下提高委内瑞拉马脑炎病毒(VEEV)、病毒衣壳蛋白和病毒基因组RNA在全血中的稳定性及检测能力。此前已证明NT颗粒能捕获并富集多种病原体,包括呼吸道合胞病毒、流感病毒、冠状病毒和裂谷热病毒。我们的研究表明,在所有温度下,与NT颗粒孵育的样本在血液中可检测到的传染性VEEV水平等于或高于未经NT处理的样本。在后期时间点(72小时)和较高温度(40°C)条件下,NT颗粒存在时病毒RNA检测增加。同样,在40°C下72小时内,NT颗粒存在时VEEV衣壳蛋白的检测增强。最后,我们用VEEV的减毒活疫苗株TC - 83经鼻感染C3H小鼠,并证明NT颗粒可显著增加在40°C下孵育长达72小时的感染血液中VEEV衣壳的检测。没有NT颗粒的样本衣壳蛋白水平无法检测到。综上所述,我们的数据证明了NT颗粒能够在一段时间内和高温下保存并实现对人及小鼠血液样本中VEEV的检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32d1/6999085/63fcd526cebd/fvets-06-00509-g0001.jpg

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