To determine the effective temperature and time for inactivation of minute virus of mice (MVM) to ensure the biosafety of the laboratory’ s samples. Through the implementation of MVM nucleic acid detection capability verification plan, we can understand the inspection capability of relevant testing institutions of laboratory animals in China, so as to improve the quality detection level and detection capability of laboratory animals in our country. Methods MVM with known virus titer was inactivated by water bath heating at different temperatures and different times. Some of the inactivated viruses were used for virus titer detection and some were used for nucleic acid detection by fluorescent PCR. The fecal suspension of MVM nucleic acid negative mice was taken as negative samples. The  inactivated virus was added to MVM nucleic acid-negative mouse fecal suspension to prepare positive samples. After passing the uniformity and stability test, the samples will be randomly numbered and distributed to the participating units. NIFDC will summarize and analyze the result submitted by the participating units. Results When the inactivation temperature reaches 100 ℃ and the inactivation time does not exceed one hour, the virus can be effectively inactivated. Meanwhile,the variation coefficient of nucleic acid test result in batches is less than 5%,and the difference is not significant. The satisfaction rate of the participants was 96. 4% ( 27 / 28) . Conclusion When the inactivation temperature reaches 100 ℃ and the inactivation time does not exceed one hour, the MVM nucleic acid sequence remains intact, and the nucleic acid detection result do not affect the experiment. There were 96. 4% of the participating laboratories had the capability of MVM nucleic acid detection, and 3. 6% needed to be improved. There are still a small number of laboratories that need to be improved in terms of operational norms and the writing of original records.

detectioncapability verification, minute virus of mice, fluorescent PCR, result evaluation

目的 确定小鼠细小病毒(MVM)有效灭活时间,保证实验室能力验证样本的生物安全性;通过 MVM 核酸检测能力验证实施计划,了解我国实验动物相关检测机构的检验能力,促进实验动物质量检测水平和能力的提高。方法 通过采取不同温度和不同时间对已知病毒滴度的 MVM 进行水浴加热灭活。 经过灭活的病毒一部分用于病毒滴度的检测,一部分通过荧光 PCR 方法用于核酸的检测。 取 MVM 核酸阴性的小鼠粪便混悬液作为阴性样品,将灭活的病毒加入到 MVM 核酸阴性的小鼠粪便混悬液中制备能力验证阳性样品。 样品经过均一性、稳定性检验合格后,采用随机编号,发放给参加单位,并对参加单位提交的结果进行汇总分析。 结果当灭活温度达 100 ℃ ,灭活时间不超过 1 h,能有效灭活病毒,时核酸检测结果批内变异系数均< 5%,差异不显著。 参加单位结果满意率为 96. 4%(27 / 28) 。 结论 当灭活温度达 100 ℃ ,灭活时间不超过 1 h,MVM 核酸序列保留完整,核酸检测结果不影响实验。 参加实验室有 96. 4%具备了 MVM 核酸检测能力,有 3. 6%的实验室能力有待提高;还有少部分实验室在操作规范和原始记录的书写规范方面有待提升。

检测能力验证, 小鼠细小病毒, 荧光 PCR 方法, 结果评价