Relova Damarys, Rios Liliam, Acevedo Ana M, Coronado Liani, Perera Carmen L, Pérez Lester J
Centro Nacional de Sanidad Agropecuaria (CENSA), OIE Collaborating Centre for Diagnosis and Risk Analysis of the Caribbean Region, La Habana 32700, Cuba.
Reiman Cancer Research Laboratory, Faculty of Medicine, University of New Brunswick, Saint John, NB E2L 4L5, Canada.
Vet Sci. 2018 Feb 6;5(1):19. doi: 10.3390/vetsci5010019.
The current global conditions, which include intensive globalization, climate changes, and viral evolution among other factors, have led to an increased emergence of viruses and new viral diseases; RNA viruses are key drivers of this evolution. Laboratory networks that are linked to central reference laboratories are required to conduct both active and passive environmental surveillance of this complicated global viral environment. These tasks require a continuous exchange of strains or field samples between different diagnostic laboratories. The shipment of these samples on dry ice represents both a biological hazard and a general health risk. Moreover, the requirement to ship on dry ice could be hampered by high costs, particularly in underdeveloped countries or regions located far from each other. To solve these issues, the shipment of RNA isolated from viral suspensions or directly from field samples could be a useful way to share viral genetic material. However, extracted RNA stored in aqueous solutions, even at -70 °C, is highly prone to degradation. The current study evaluated different RNA storage conditions for safety and feasibility for future use in molecular diagnostics. The in vitro RNA-transcripts obtained from an inactivated highly pathogenic avian influenza (HPAI) H5N1 virus was used as a model. The role of secondary structures in the protection of the RNA was also explored. Of the conditions evaluated, the dry pellet matrix was best able to protect viral RNA under extreme storage conditions. This method is safe, cost-effective and assures the integrity of RNA samples for reliable molecular diagnosis. This study aligns with the globally significant "Global One Health" paradigm, especially with respect to the diagnosis of emerging diseases that require confirmation by reference laboratories.
当前的全球状况，包括高度全球化、气候变化以及病毒进化等诸多因素，导致病毒和新型病毒性疾病的出现日益增多；RNA病毒是这一进化过程的关键驱动因素。需要与中央参考实验室相连的实验室网络，对这种复杂的全球病毒环境进行主动和被动的环境监测。这些任务要求不同诊断实验室之间持续交换毒株或现场样本。使用干冰运输这些样本既存在生物危害，也有一般健康风险。此外，使用干冰运输的要求可能会因成本高昂而受阻，特别是在不发达国家或彼此距离遥远的地区。为解决这些问题，运输从病毒悬液中分离或直接从现场样本中提取的RNA，可能是共享病毒遗传物质的一种有效方式。然而，即使保存在-70°C的水溶液中的提取RNA也极易降解。本研究评估了不同RNA储存条件的安全性和可行性，以便未来用于分子诊断。从灭活的高致病性禽流感（HPAI）H5N1病毒获得的体外RNA转录本被用作模型。还探讨了二级结构在保护RNA中的作用。在所评估的条件中，干颗粒基质在极端储存条件下最能保护病毒RNA。该方法安全、经济高效，并能确保RNA样本的完整性以进行可靠的分子诊断。本研究符合具有全球重要意义的“全球一体化健康”范式，特别是在诊断需要参考实验室确认的新发疾病方面。
