诺如病毒检验用大肠杆菌噬菌体MS2标准物质的研制

Preparation of coliphage MS2 reference material for norovirus test

中文关键词:  诺如病毒  微生物标准物质  大肠杆菌噬菌体MS2  冷冻干燥

英文关键词:norovirus  microbial reference material  coliphage MS2  freeze-drying

中文摘要:

      目的 制备均匀性和稳定性符合要求的大肠杆菌噬菌体MS2标准物质, 用于诺如病毒检验过程质量控制。方法 对大肠杆菌噬菌体MS2 (CMCCPh001)进行分子生物学确认。采用冷冻干燥技术制成微球状标准物质, 并进行均匀性、长期稳定性、短期稳定性检验。以牡蛎等5种样品作为基质, 对使用效果进行确认。 结果 制备的样品呈球形、白色、大小均一, 其均匀性符合要求, 经?20 ℃ 12个月长期储存, 样品稳定, 经4 ℃ 28 d以及37 ℃ 5 d的短期储存, 样品稳定。阈值循环数(threshold cycle, Cq)值与均匀性检平均值差异不大。牡蛎等5种样品使用效果确认实验表明, 该标准物质可以作为食品诺如病毒检验过程质量控制使用。结论 大肠杆菌噬菌体MS2 (CMCCPh001)分子水平鉴定结果符合大肠杆菌噬菌体MS2特征。均匀性、长期稳定性、短期稳定性及适用性实验结果符合作为诺如病毒检验过程质量控制标准物质的要求。

英文摘要:

      Objective To prepare coliphage MS2 reference material with required uniformity and stability, which can be used for the quality control of norovirus detection process. Methods The coliphage MS2 (CMCCPh001) was confirmed by molecular biology method. The microsphere-shaped reference material was prepared by freeze-drying technology, and the homogeneity, long-term stability and short-term stability were tested. Five kinds of samples including oyster were used as the matrixes to confirm the using effects. Results The prepared samples were spherical, white and uniform in size, and their uniformity met the requirements. After long-term storage at ?20 ℃ for 12 months, the samples were stable. After short-term storage at 4 ℃ for 28 d and 37 ℃ for 5 d, the samples were stable. There was no significant difference between the threshold cycle (Cq) value and the average value of uniformity test. The validation experiment of 5 kinds of samples such as oyster showed that the reference material could be used as the quality control of food norovirus detection process. Conclusion The molecular level identification result of coliphage MS2 (CMCCPh001) is consistent with the characteristics of coliphage MS2. The uniformity, long-term stability, short-term stability and applicability test results meet the requirements for quality control reference materials in the norovirus inspection process.