Guangdong Provincial Key Laboratory of Modern Geometric and Mechanical Metrology Technology, Guangdong Provincial Institute of Metrology, Guangzhou,Guangdong 510405, China
Abstract：The bacterial colony itself is unstable and inconvenient to carry, so it is not suitable for use as a reference material for calibration of colony counters. Colony simulation images were used to represent colonies. According to the characteristics of colonies, colony images of different sizes and uneven distribution were processed into pictures, and the total number and size of simulated colonies were simulated to simulate standard colony materials. The total number and size of the simulated bacterial colony were equivalent to the simulated bacterial colony reference materials.A stratified random sampling method was used to study the homogeneity and stability of the reference materials.The results showed that the prepared reference materials had good homogeneity and stability.Through rigorous calculation and data processing, the total number and size of the simulated bacterial colony were determined and the uncertainty was calculated.The series of reference materials could be stored and carried easily, so they were suitable for bacterial colony counter calibration and analysis method evaluation, and have been certified as the secondary national certified reference materials with the number of GBW(E) 130586~GBW(E) 130590.
［1］隋志伟, 刘晓夏, 王晶, 等.粘质沙雷氏菌测量方法研究［J］.计量学报, 2018,39(4):588-592.
Sui Z W, Liu X X, Wang J. et al. Study on Measurement Method of Serratia Marcescens［J］.Acta Metrologica Sinica, 2018, 39(4): 588-592.
［2］刘明爽, 李婷婷, 马艳, 等. 真空包装鲈鱼片在冷藏与微冻贮藏过程中的新鲜度评价［J］. 食品科学, 2016, 37(2): 210-213.
Liu M S, Li T T, Ma Y, et al. Freshness Evaluation of Vacuum Packaged Perch Fillets during Refrigeration and Partial Freezing［J］.Food Science, 2016, 37(2): 210-213.
［3］卢行安, 顾其芳, 袁宝君, 等. AOAC Petrifilm~(TM)菌落总数测试片法与食品中菌落总数测定国标方法的比较研究［J］. 中国食品学报, 2011, 11(3): 164-167.
Lu X A, Gu Q F, Yuan B G, et al. The Comparing Study of AOAC Petrifilm~(TM) Aerobic Count Plate Methods with GB Aerobic Plate Count in Foods［J］. Journal of Chinese Institute of Food Science and Technology, 2011, 11(3): 164-167.
［4］Loss G, Apprich S, Kneifel W, et al. Short communication: Appropriate and alternative methods to determine viable bacterial counts in cow milk samples［J］. Journal of Dairy Science, 2012, 95(6): 2916-2918.
［5］刘学铭, 方少钦, 唐道帮, 等. 我国熟肉制品微生物安全现状与控制技术［J］. 现代食品科技, 2012, 28(1): 99-103.
Liu X M, Fang S Q, Tang D B, et al. Microbiological Safety and Control Technology of Cooked Meat Products in China［J］. Modern Food Science and Technology, 2012, 28(1): 99-103.
［6］杨宁, 张荣标, 赵雨琦, 等. 基于荧光显微拉曼光谱的细菌快速解耦检测方法［J］. 仪器仪表学报, 2012, 33(2): 435-441.
Yang N, Zhang R B, Zhao Y Q, et al. Bacteria fast decoupling detection based on Raman spectra and fluorescence microscopy［J］. Chinese Journal of Scientific Instrument, 2012, 33(2): 435-441.
［7］陶斐斐, 王伟, 李永玉, 等. 冷却猪肉表面菌落总数的快速无损检测方法研究［J］. 光谱学与光谱分析, 2010, 30(12): 3405-3409.
Tao F F, Wang W, Li Y Y, et al. A Rapid Nondestructive Measurement Method for Assessing the Total Plate Count on Chilled Pork Surface［J］. Spectroscopy and Spectral Analysis, 2010, 30(12): 3405-3409.
［8］夏宏丽. 使用标准物质评定食品菌落总数的不确定度［J］. 中国卫生检验杂志, 2012,(6): 1450-1451.
Xia H L. Evaluation of Uncertainty of Total Colony Number in Food by Reference Materials［J］. Chinese Journal of Health Laboratory Technology, 2012, (6): 1450-1451.
［9］王海华, 兰茜. 能力验证菌落总数测定结果不确定度的评定［J］. 食品安全质量检测学报, 2015,(6): 2352-2355.
Wang H H, Lan Q, Uncertainty Evaluation of Aerobic Plate Count by Proficiency Testing［J］. Journal of Food Safety and Quality, 2015, (6): 2352-2355.
［10］Veld P H I, Havelaar A H, Strijp-Lockefeer N G W M V. The certification of a reference material for the evaluation of methods for the enumeration of Bacillus cereus［J］. Journal of Applied Microbiology, 2010, 86(2): 266-274.
［11］白凤翎. 国内外食品卫生微生物标准菌落总数的比较研究［J］. 食品科技, 2008, 33(3): 197-199.
Bai F L. Study on ABC of microbiological criteria of food hygiene between national and abroad［J］. Food Science and Technology, 2008, 33(3): 197-199.
［12］陈敏璠, 傅博强, 林婧, 等. 阴沟肠杆菌定量标准物质的研制［J］. 计量学报, 2017, 38(5): 527-531.
Chen M F, Fu B Q, Lin J. et al. Development of Quantitative Reference Material of Enterobacter Cloacae［J］. Acta Metrologica Sinica, 2017, 38(5): 527-531.
［13］吴玉峰, 丰丽艳. 全自动菌落分析仪计数结果的初步探讨［J］. 环境卫生学杂志, 2013, (5): 420-421.
Wu Y F, Feng L Y. Preliminary Study on the Results of an Automatic Colony Counts Analyzer［J］. Journal of Environmental Hygiene, 2013, (5): 420-421.
［14］张春晨, 杨更发.分值计算法应用于培养基质量评价及菌落计数的质控［J］.预防医学情报杂志, 1999,(2): 109-110.
Zhang C C, Yang G F. Application of Score Calculating Method in Quality Evaluation of Culture Medium and Quality Control of Colony Count［J］. Journal of Preventive Medicine Information, 1999,(2): 109-110.
［15］蒋峰, 罗黎明, 陈建军, 等. 基于改进的Hough算法的细胞破碎智能监测计数系统［J］. 计量学报, 2018, 39(6): 902-907.
Jiang F, Luo L M, Chen J J, et al. Research of Cell Lysis Intelligent Monitoring and Counting System Based on the Improved Hough Algorithm［J］. Acta Metrologica Sinica, 2018, 39(6): 902-907.
［16］Viitanen H, Vinha J, Salminen K, et al.Moisture and Bio-deterioration Risk of Building Materials and Structures［J］. Journal of Building Physics, 2010, 33(3): 201-224.
［17］GB 4789.2—2016食品微生物菌落总数的测定［S］. 2016.
［18］JJF 1006—1994 一级标准物质技术规范［S］.1994.
［19］ISO Guide 34 General requirements for the competence of reference material producers［S］.2009.
［20］ISO Guide 35 Reference materials-General and statistical principles for certification［S］.2006.
2020, Vol. 41 
