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Research Project: Viral Ecology of Henipaviruses in Endemic Settings and Intervention Strategies to Prevent their Spread to Domestic Animals

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Title: Effect of temperature and time on RNA detection by RT-qPCR in rodent tissue and blood samples stored in MagMAX™ lysis/binding solution concentrate: Considerations for RNA detection in specimens stored under suboptimal

Author
item DAVIES, KATHERINE - Oak Ridge Institute For Science And Education (ORISE)
item WELCH, STEPHEN - Centers For Disease Control And Prevention (CDC) - United States
item SORVILLO, TERESA - Centers For Disease Control And Prevention (CDC) - United States
item COLEMAN-MCCRAY, JOANN - Centers For Disease Control And Prevention (CDC) - United States
item SPIROPOULOU, CHRISTINA - Centers For Disease Control And Prevention (CDC) - United States
item SPENGLER, JESSICA - Centers For Disease Control And Prevention (CDC) - United States

Submitted to: Journal of Virological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/6/2025
Publication Date: 5/8/2025
Citation: Davies, K.A., Welch, S.R., Sorvillo, T.E., Coleman-Mccray, J., Spiropoulou, C.F., Spengler, J.R. 2025. Effect of temperature and time on RNA detection by RT-qPCR in rodent tissue and blood samples stored in MagMAX™ lysis/binding solution concentrate: Considerations for RNA detection in specimens stored under suboptimal conditions. Journal of Virological Methods. 337. Article 115175. https://doi.org/10.1016/j.jviromet.2025.115175.
DOI: https://doi.org/10.1016/j.jviromet.2025.115175

Interpretive Summary: Tissue and blood samples, containing or potentially containing infectious material must be inactivated before downstream analysis at lower containment e.g., molecular analysis. Storage in traditional methods (such as liquid nitrogen flash freezing) may not be available in resource-limited settings, additionally buffers like RNAlater may provide high levels of RNA preservation but has been demonstrated to also preserve viral infectivity. Given the biohazard risks associated with sample collection, transportation, and analysis with high consequence pathogens, the option to store samples directly in a known viral inactivant can reduce overall exposure risk. Here we demonstrate temporary storage in MagMAX lysis buffer can provide logistical options for those working both in the field and in laboratory settings where immediate processing of samples may be challenging or when unexpected challenges like cold storage failures or delays in sample processing may occur. Our data demonstrates samples can be refrigerated for up to one year and stored at warmer temperatures for two weeks (up to 32°C) or twelve weeks (at room temperature) without compromising RNA detection.

Technical Abstract: Tissue samples are often collected into sample lysis buffers prior to downstream molecular analysis. Immediate sample processing and cold storage are not always possible during large-scale or field studies, or in facilities lacking stable electrical supply. Additionally, samples may need to be transported significant distances before processing. Here, we investigate the long-term stability and detection of RNA in tissues and blood stored in MagMAX lysis media for up to a year at -80°C, 4°C, 21°C, and 32°C.