Cell-free debris fraction was diluted 1:10 in medium before combination with ViroMag reagent. Infection in the ViroMag containing wells were comparable to infection with 10-fold higher concentrated virus with better shape and viability.
This paper shows the efficiency of ViroMag from OZ Biosciences to enhance infectiveness of VZV virus.article reference: J Virol Methods. 2014 Jun 9. pii: S0166-0934(14)00219-5
Laboratory preparation of varicella zoster virus: Concentration of virus-containing supernatant, use of a debris fraction and magnetofection for consistent cell-free VZV infections.
Sloutskin A, Goldstein RS.AbstractThe research laboratory generation of free varicella-zoster virus (VZV) from cultured cells results generally in relatively low titers, with the result that most study of VZV infection utilizes cell-associated infection. VZV modified genetically to express fluorescent proteins have proven a very useful tool in the study of the cell biology of the virus. However, important aspects of VZV-cell interaction, such as the entry mechanism and superinfection exclusion have not yet been studied in detail, in part due to the difficulty in obtaining a high titer cell free virus. Here, a method to generate relatively high-titer cell-free VZV, based on a combination of previously published techniques and subsequent concentration is described. VZV-infected cells are disrupted, sonicated, clarified by centrifugation and PEG concentrated. The generated cell-free virus in the supernatant is then concentrated to yield up to 105PFU/ml. The cell debris pellet, which contains up to 106PFU/ml can also be used. Magnetic nanoparticles available commercially can be used to enhance infection by cell-free-VZV. The tools described here hold promise for better understanding of important aspects of VZV-cell interactions such as entry and latency.
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