检测能力验证, 小鼠细小病毒, 荧光 PCR 方法, 结果评价

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