Obtaining RNA control sample and prospects of its application for diagnostics of yellow fever by the LAMP method
https://doi.org/10.31677/2072-6724-2025-76-3-261-268
Abstract
The aim of the study was to create a control RNA sample for yellow fever virus (YFV) diagnostics using the LAMP method. Two control samples were obtained: recombinant plasmid DNA (pCR2.1-YFV-K+) and mRNA (RNA-YFV-K+). During the assessment of the analytical sensitivity by the LAMP method, it was found that both samples demonstrated a comparable detection limit of 104 copies/ml, which allows for effective detection of the viral genetic material in the first days of disease symptoms. A study of the sample stability showed that at a temperature of -15 to -25 °C, both samples retain their functionality even after tenfold defrosting/freezing. At a temperature of 2 to 8 °C, the samples are determined as positive for 7 days of storage. However, stability decreases at room temperature: for pCR2.1-YFV-K+ it remains up to 72 hours, and for RNA-YFV-K+ — up to 48 hours. To ensure stability, samples should be stored at low temperatures (-15 to -25°C or 2 to 8°C). Based on the results, RNA-YFV-K+ can be used as a control for analytical sensitivity and correctness of the reverse transcription reaction followed by amplification by the LAMP method, which is important for the detection of RNA-containing yellow fever virus.
About the Authors
E. I. Krivosheina
State Scientific Center of Virology and Biotechnology «Vector»
Russian Federation
Russian Federation
Research Associate.
Kol’tsovo
K. R. Belichenko
State Scientific Center of Virology and Biotechnology «Vector»
Russian Federation
Russian Federation
Laboratory Research Assistant.
Kol’tsovo
E. V. Chub
State Scientific Center of Virology and Biotechnology «Vector»
Russian Federation
Russian Federation
PhD in Biology, Leading Researcher.
Kol’tsovo
M. Yu. Kartashov
State Scientific Center of Virology and Biotechnology «Vector»
Russian Federation
Russian Federation
PhD in Biology, Senior Researcher.
Kol’tsovo
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Review
For citations:
Krivosheina E.I., Belichenko K.R., Chub E.V., Kartashov M.Yu. Obtaining RNA control sample and prospects of its application for diagnostics of yellow fever by the LAMP method. Bulletin of NSAU (Novosibirsk State Agrarian University). 2025;(3):261-268. (In Russ.) https://doi.org/10.31677/2072-6724-2025-76-3-261-268
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