Marthe Colotte , Aurélie Luis , Delphine Coudy , Sophie Tuffet , Isabelle Robene , Babbitha Fenelon , Emmanuel Jouen , Nicolas Leveque , Luc Deroche , Sophie Alain , Dorian Plumelle , Camille Tumiotto , Laurent Busson , Marie-Edith Lafon , Jacques Bonnet
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引用次数: 0
Abstract
The Coronavirus pandemic unveiled the unprecedented need for diagnostic tests to rapidly detect the presence of pathogens in the population. Real-time RT-PCR and other nucleic acid amplification techniques are accurate and sensitive molecular techniques that necessitate quality control strategies and stable quality control materials. To meet this need, Twist Bioscience has developed and released synthetic RNA controls. However, RNA is an inherently unstable molecule needing cold storage, costly shipping, and resource-intensive logistics. Imagene provides a solution to this problem by encapsulating dehydrated RNA inside metallic capsules filled with anhydrous argon, allowing room temperature and eco-friendly storage and shipping. This technology initially developed for DNA storage has been successfully applied to RNA and other biospecimen and extensively validated through real time and accelerated aging. Here, RNA controls produced by Twist Bioscience were encapsulated in RNAshells and distributed to several laboratories that used them for COVID-19 detection tests by amplification. One RT-LAMP procedure, four different RT-PCR devices and 6 different PCR kits were used. The amplification targets were genes E, N; RdRp, Sarbeco-E and Orf1a/b. RNA retrieval was satisfactory, and the detection was reproducible. RNA stability was checked by real-time (3 years at room temperature) and accelerated aging (16 h at 90 °C, corresponding to approximately 10 years of storage at 25 °C, according to our previously published Arrhenius study for encapsulated RNA). The results were not significantly different from those for unaged capsules. This room temperature RNA stability allows the preparation and distribution of large strategic batches which can be stored for more than 10 years a long time and used for standardization processes between detection sites. Moreover, this provides the advantage of single-use and field usability across varying temperatures. Consequently, this type of encapsulated synthetic RNA, processed at room temperature, can be used as internal quality control materials for the SARS-Cov-2 virus as well as for detection of other RNA viruses.
期刊介绍:
The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery.
The methods may include, but not limited to, the study of:
Viral components and morphology-
Virus isolation, propagation and development of viral vectors-
Viral pathogenesis, oncogenesis, vaccines and antivirals-
Virus replication, host-pathogen interactions and responses-
Virus transmission, prevention, control and treatment-
Viral metagenomics and virome-
Virus ecology, adaption and evolution-
Applied virology such as nanotechnology-
Viral diagnosis with novelty and comprehensive evaluation.
We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.