Visualizing the Transport of Porcine Reproductive and Respiratory Syndrome Virus in Live Cells by Quantum Dots-Based Sin
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RESEARCH ARTICLE
Visualizing the Transport of Porcine Reproductive and Respiratory Syndrome Virus in Live Cells by Quantum Dots-Based Single Virus Tracking Zhenpu Liang1 • Pengjuan Li1 • Caiping Wang1 • Deepali Singh2 • Xiaoxia Zhang1 Received: 20 August 2019 / Accepted: 9 October 2019 Ó Wuhan Institute of Virology, CAS 2019
Abstract Quantum dots (QDs)-based single particle analysis technique enables real-time tracking of the viral infection in live cells with great sensitivity over a long period of time. The porcine reproductive and respiratory syndrome virus (PRRSV) is a small virus with the virion size of 40–60 nm which causes great economic losses to the swine industry worldwide. A clear understanding of the viral infection mechanism is essential for the development of effective antiviral strategies. In this study, we labeled the PRRSV with QDs using the streptavidin–biotin labeling system and monitored the viral infection process in live cells. Our results indicated that the labeling method had negligible effect on viral infectivity. We also observed that prior to the entry, PRRSV vibrated on the plasma membrane, and entered the cells via endosome mediated cell entry pathway. Viruses moved in a slow–fast–slow oscillatory movement pattern and finally accumulated in a perinuclear region of the cell. Our results also showed that once inside the cell, PRRSV moved along the microtubule, microfilament and vimentin cytoskeletal elements. During the transport process, virus particles also made contacts with non-muscle myosin heavy chain II-A (NMHC II-A), visualized as small spheres in cytoplasm. This study can facilitate the application of QDs in virus infection imaging, especially the smaller-sized viruses and provide some novel and important insights into PRRSV infection mechanism. Keywords Single virus tracking Quantum dots (QDs) Biotinylation Porcine reproductive and respiratory syndrome virus (PRRSV) Transport
Introduction Viruses are the one of the most ancient life forms, and causative agents of approximately 70% diseases of human and animals. Studies related to the invasion and reproduction mechanism of viruses provide significant insight into the virus life cycle, which can be later exploited for the Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12250-019-00187-0) contains supplementary material, which is available to authorized users. & Xiaoxia Zhang [email protected] 1
College of Life Sciences, Key Laboratory of Enzyme Engineering of Agricultural Microbiology (Ministry of Agriculture), Henan Agricultural University, Zhengzhou 450000, China
2
School of Biotechnology, Gautam Buddha University, Greater Noida 201312, India
development of drugs and therapeutics against viral diseases (Simmonds and Domingo 2011). Viral infections in cells are visualized by immunofluorescence which requires the infected cell to be fixed using paraformaldehyde, therefore immunoflorescence
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