产气荚膜梭菌质控样品的研制  

Preparation of the quality control samples for Clostridium perfringens

作　　者：任敏[1] 刘悦[1] 王秋水 邓婕 马凯[1] 左嘉[1] 袁立艳 高丽娟[1] REN Min;LIU Yue;WANG Qiu-Shui;DENG Jie;MA Kai;ZUO Jia;YUAN Li-Yan;GAO Li-Juan(Beijing Center for Physical&Chemical Analysis,Beijing Engineering Research Center of Food Safety Analysis,Beijing 100089,China)

机构地区：[1]北京市理化分析测试中心,北京市食品安全分析测试工程技术研究中心,北京100089

出　　处：《食品安全质量检测学报》2021年第10期4274-4279,共6页Journal of Food Safety and Quality

基　　金：北京市科学技术研究院改革发展培育项目(PY2020JK44);北京市科学技术研究院市级财政项目(PXM2020178305000007)。

摘　　要：目的制备产气荚膜梭菌质控菌株质控样品。方法将产气荚膜梭菌菌悬液于36℃厌氧培养至浓度为107 CFU/mL,以10%(m/m,下同)脱脂奶粉、5%海藻糖、5%蔗糖、1%谷氨酸钠、0.5%抗坏血酸钠为菌体保护剂,使用冷冻干燥法制备冻干菌剂。依据CNAS-CL03-A001:2019《能力验证提供者认可准则在微生物领域的应用说明》,通过均匀性、运输稳定性、储藏稳定性等指标对制备出的菌剂进行评价。结果在均匀性实验中样品F值为3.00,小于F临界值F0.05(9,10)=3.02;模拟运输稳定性实验中,于4、15和25℃温度下模拟运输5 d,检验t值均小于临界值t0.05(24)=2.0639;在储藏稳定性实验中,于4、-20和-80℃下储藏28 d内,检验t值均小于临界值t0.05(24)=2.0639,都符合标准要求。结论制备的产气荚膜梭菌质控菌株均匀性、储藏稳定性和运输稳定性良好,可以作为实验室质控样品使用。Objective To prepare quality control samples of Clostridium perfringens.Methods The suspension of Clostridium perfringens was anaerobicly cultured at 36℃to a concentration of 107 CFU/mL.Using 10%(m/m,the same below)skim milk powder,5%trehalose,5%sucrose,1%sodium glutamate and 0.5%sodium ascorbate as bacterial protective agents,the lyophilized bacterial agent was prepared by freeze-drying method.According to CNAS-CL03-A001:2019 Specification for application of accreditation criteria for proficiency testing providers in the field of microbiology,the prepared microbial agents were evaluated by the uniformity,transportation stability,storage stability and other indicators.Results In the uniformity experiment,the F value of the sample was 3.00,which was less than the critical F value F0.05(9,10)=3.02.In the simulation transportation stability experiment,the simulated transportation was carried out at 4,15 and 25℃for 5 days,and the test t value of the sample was less than the critical value t0.05(24)=2.0639.In the storage stability experiment,within 28 days of storage at 4,-20 and-80℃,the test t value was less than the critical value t0.05(24)=2.0639,which all met the standard requirements.Conclusion The prepared quality control strain of Clostridium perfringens has good uniformity,storage stability and transportation stability,and can be used as a laboratory quality control sample.

关 键 词：产气荚膜梭菌 质控样品 冷冻干燥 

分 类 号：TS201.3[轻工技术与工程—食品科学]