During a foot-and-mouth disease (FMD) outbreak, transport and testing of potentially infectious samples, including epithelium from suspect lesions, presents a biosafety risk, particularly in FMD-free countries. Therefore, treatment to inactivate virus prior to transport is important. Tongue epithelium from cattle infected with FMD virus (FMDV) serotype O (O ALG/3/2014 - Lineage O/ME-SA/Ind-2001d) or A (A IRN/22/2015 - Lineage A/ASIA/G-VII) was incubated in RNAlater, RNA Shield or phosphate-buffered saline (pH 7.4) at room temperature for 2, 6, 24 or 48 h. After incubation, tissues were homogenised and tested by virus titration. Viral RNA in the homogenate was quantified by RT-qPCR, used for sequencing, and transfected into LFBKαVβ6 cells to recover infectious virus. RNAlater reduced A IRN/22/2015 titres by 4 log10 after 24 h, and completely after 48 h incubation. While O ALG/3/2014 was detected by VI after 2, 6 and 24 h, titration yielded no infectious virus, likely as a result of freeze-thawing. RNA Shield was cytotoxic at high concentrations but was effective at inactivating both strains after 24 h. Regardless of reagent or inactivation period, RT-qPCR, VP1 sequencing, and transfection of RNA to recover infectious virus were possible. RNA Shield appears a better choice for FMDV inactivation in tissues, however 24 h incubation is recommended.

中文翻译：

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灭活上皮样品中的口蹄疫病毒，以便在低浓度实验室中安全运输和加工。

在口蹄疫（FMD）爆发期间，潜在传染性样品（包括可疑病变的上皮）的运输和测试存在生物安全风险，尤其是在无FMD的国家/地区。因此，在运输之前使病毒灭活的治疗很重要。来自感染了口蹄疫病毒（FMDV）血清型O（O ALG / 3/2014-血统O / ME-SA / Ind-2001d）或A（A IRN / 22/2015-血统A / ASIA / G-VII）的牛的舌上皮）在室温下于RNAlater，RNA Shield或磷酸盐缓冲盐水（pH 7.4）中孵育2、6、24或48 h。孵育后，将组织匀浆并通过病毒滴定进行测试。通过RT-qPCR定量匀浆中的病毒RNA，用于测序，并转染至LFBKαVβ6细胞中以回收感染性病毒。24小时后，RNAlater将A IRN / 22/2015滴度降低了4 log10，并在孵育48小时后完全清除。虽然在2、6和24小时后通过VI检测到O ALG / 3/2014，但滴定没有产生传染性病毒，这可能是冻融的结果。RNA Shield在高浓度下具有细胞毒性，但在24小时后可有效灭活这两种菌株。无论试剂或灭活时间长短，都可以进行RT-qPCR，VP1测序和RNA转染以恢复感染性病毒。RNA Shield似乎是组织中FMDV失活的更好选择，但是建议进行24小时孵育。RNA的转染以恢复感染性病毒是可能的。RNA Shield似乎是组织中FMDV失活的更好选择，但是建议进行24小时孵育。RNA的转染以恢复感染性病毒是可能的。RNA Shield似乎是组织中FMDV失活的更好选择，但是建议进行24小时孵育。