Recovery of live virus after storage at ambient temperature using ViveST™
Section snippets
Background
The most accurate approaches to viral characterization require the collection of viable virus which is amplified in culture. Often, field collection and preservation of circulating virus requires ultra-low freezers or dry ice. Because such cold-chain infrastructure is not available in many developing countries, viral characterization data in these countries can be sparse. Even if virus can be successfully preserved and shipped from the field site to a reference laboratory, there are often long
Objective
Using in vitro viral titer studies, we sought to evaluate the ability of the ViveST device to preserve live virus at ambient temperature in comparison with other viral transport media.
Viruses
A panel of six viruses was used in this study in order to determine the range of viruses that could be preserved with ViveST. Viruses of various sizes were evaluated and included those with DNA or RNA genomes, with or without envelopes. This panel included enteric cytopathic human orphan virus 3 (ECHOV), Morrisey; human rhinovirus 15 (HRV), 1734; human Coxsackievirus B5 (CBV), Faulkner and herpes simplex virus 1 (HSV), HF. These viruses are prototype strains of the Center for Disease Control
ViveST preserves virus at ambient temperature for 7 days
Viable HAdV, CBV, ECHOV, HSV, and HRV were recovered from the ViveST with FBS treatment after 7 days storage at ambient temperature (Fig. 2). Viruses showed an average reduction in infectious units (i.u.) by 2 orders of magnitude when compared to the starting titer of the respective virus (Fig. 2). ECHOV exhibited the highest rate of preservation of i.u. when compared with HAdV, HSV, and HRV (p = 0.00006). While there was a significant loss of i.u. for all viruses (p = 0.00035), at least 15,000
Discussion
Currently there are no standard methods for the storage and recovery of live virus at ambient temperature and investigations in this subject are sparse. Studies investigating the use of cell culture media and viral transport media for preserving virus at room temperature have documented a low recovery rate for HSV, HAdV, and other viruses tested.4, 5 Recovery of live HIV from ViveST has been attempted, but was unsuccessful using the standard drying protocol.12 This may have been a result of the
Funding
This work was supported by grants from the US Department of Defense Armed Forces Health Surveillance Center's Global Emerging Infections Surveillance and Response Program (I0179_10_UN Dr. Gray) and from the National Institute of Allergy and Infectious Diseases (R01 AI068803 – Dr. Gray).
Competing interest
None declared.
Ethical approval
Not required.
Acknowledgments
We thank Mimi Healy of Vivebio, for her support, advice, and encouragement concerning this work. We are also grateful to Thomas Waltzek for his insights and recommendations on viruses.
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