大肠埃希氏菌O157∶H7/NM标准物质制备  

The Reference Materials of Preparation About Escherichia Coli O157∶H7/NM

作　　者：柯璐[1,2] 戴晓丽 徐珊[1,2] 黄嫦娇 林杰[1,2] KE Lu;DAI Xiaoli;XU Shan;HUANG Changjiao;LIN Jie(Fujian Entry-exit Inspection and Quarantine Bureau,Fuzhou,Fujian,350003,China;Fujian Key Laboratory for Technology Research of Inspection and Quarantine)

机构地区：[1]福建出入境检验检疫局技术中心,福建福州350003 [2]福建省检验检疫技术研究重点实验室

出　　处：《检验检疫学刊》2019年第6期10-17,共8页Journal of Inspection and Quarantine

基　　金：原福建出入境检验检疫局科技计划项目(FK2016-JS001)

摘　　要：本文用冷冻干燥法制备含鸡肉基质的大肠埃希氏菌O157∶H7/NM标准物质,以细菌存活率为指标,选择存活率最高的保护剂配方。以经钴-60辐照灭菌的鸡肉粉(无抗生素)作为基质,按适当的比例与保护剂配方液混合,添加菌液,于-70℃速冻20 h后冷冻干燥,对样品进行均匀性检测、稳定性追踪以及建立不确定度评估。结果显示,保护剂最优配方为10%海藻糖+25%脱脂牛奶+4%聚乙烯吡咯烷酮+0.8%甘氨酸;基质液比例为基质∶水=1∶7。按基质液:保护剂配方液=1∶3制备样品,均匀性试验:F统计量=1.197<2.201,均匀性良好。当测量检验结果用2次测量值平均值表示结果时,其值分布区间为±80 CFU/mL,且在-20℃的温度下保存18个月未观测到不稳定性。This experiment studies the method of the Escherichia coli O157:H7/NM reference materials containing chicken substrate by the freeze-drying preparation. For indicators of bacterial survival rate to choose freeze-dried protection scheme of highest protectant formula. The chicken(antibiotic free) powder sterilizated by Cobalt-60 irradiation as substrate, mixed with compound protective agent, adding bacteria liquid. Pre freezing at-70℃ for 20 h and freeze-drying. Test the uniformity stability, tracking, and assessment of uncertainty of reference material.The results show that the optimal formulation of the protectant was 10% trehalose+25% skimmed milk+4% PVP+0.8% glycine;the ratio of matrix solution to water is 1 ∶7;the samples are prepared according to the ratio of matrix solution to protectant solution.Uniformity test: F statistic=1.197<2.201, good uniformity. When the results of measurement and inspection are represented by the average of two measurements, the value distribution range is between ±80 CFU/mL. Stable is observed at-20℃ for 18 months.

关 键 词：大肠埃希氏菌O157:H7/NM 标准物质 冷冻干燥法 保护剂 

分 类 号：TB99[一般工业技术—计量学]