7 7 Vol. 7 No. 7 2016 7 Journal of Food Safety and Quality Jul., 2016 * 钟玮, 张宁, 刘玉萍, 范一灵, 杨美成 (, 201203) 摘要 : 目的 方法, (RiboPrinter ) 结果 CODE 1~9, CODE 10 Z 2; 10 CFU/g, 10 结论 关键词 : ; ; ; ; Proficiency testing results and analysis of quantitative detection of Staphylococcus aureus in food ZHONG Wei, ZHANG Ning, LIU Yu-Ping, FAN Yi-Ling, YANG Mei-Cheng * (Shanghai Institute for Food and Drug Control, Shanghai 201203, China) ABSTRACT: Objective To improve the detection capacity of Staphylococcus aureus in food. Methods Detection was followed the instruction of the proficiency testing, and RiboPrinter was used for identification and ribotyping of the bacteria added into the blind sample. Results Staphylococcus aureus was detected in CODE 1~9 and there was no Staphylococcus aureus in CODE 10. The Z scores of the positive samples were all less than 2 and the result of the negative sample was less than 10 CFU/g. The test results of 10 samples were satisfied. Conclusion The laboratory detection capability has been improved by implementing proficiency testing. KEY WORDS: Staphylococcus aureus; quantitative; proficiency testing; identification; ribotyping 1 引言 (Staphylococcus aurues, S. aurues),,,,, 30%~80%, [1] [2],,, [3], 2015 * 通讯作者 :,,, E-mail: yangmeicheng@vip.sina.com *Corresponding author: ANG Mei-Cheng, Ph.D., Chief Pharmacist, Shanghai Institute for Food and Drug Control, Shanghai 201203, China. E-mail: yangmeicheng@ vip.sina.com
7, : 2759 [4], 2 材料与方法 2.1 材料 试剂和仪器, 10, CODE 1~10, ; 10, 25 g, CODE 1~10 1 (, ); Baird-Parker ( BD ); ( ); ( ); - (BHI)( ); ( ); ( ); RiboPrinter ( Qualicon ) RiboPrinter ( Qualicon ) 2.2 实验方法 2.2.1 金黄色葡萄球菌定量检验,, 225 ml,,, 10 GB4789.10-2010 Baird-Parker [5],, 10, 1 ml 0.3 0.3 0.4 ml Baird-Parker, L,, (36±1) 45~48 h,, GB 4789.10-2010 Baird-Parker, 2 GB 4789.10-2010, Baird-Parker,, 3 20~200 CFU, [5] 2.2.2 盲样中添加菌的鉴定及分型 (1) [6] EcoR Ⅰ, RiboPrinter, 40 μl, 30μL, 5 μl A B, RiboPrinter,, DuPontID, (2) RiboPrinter BioNumerice (Version 6.6), Pearson, (unweighted pair-group method with arithmetic means, UPGMA), 3 结果与讨论 3.1 金黄色葡萄球菌定量检测 10, 3 1, Baird-Parker,, [5], -20,, Baird-Parker, 10,,,, --, GB4789.10-2010, 3, Baird-Parker,,, 48 h,, [7], ;,,,, 18~24 h, ;, [8] Baird-Parker,, [9,10],,, [11]
2760 7 表 1 金黄色葡萄球菌定量检测平板计数结果 Table 1 The counting results of S. aureus 10-1 10-2 10-3 10-4 (CFU/g) Baird-Parker 300 34 0 0 3.4 10 3 CODE-1 300 28 0 0 2.8 10 3 300 34 5 1 3.4 10 3 Baird-Parker 272 22 0 0 2.2 10 3 CODE-2 185 18 0 0 1.9 10 3 300 36 4 0 3.6 10 3 Baird-Parker 300 47 0 0 4.7 10 3 CODE-3 300 45 0 0 4.5 10 3 300 54 5 0 5.4 10 3 Baird-Parker 300 41 4 0 4.1 10 3 CODE-4 300 38 0 0 3.8 10 3 300 49 5 0 4.9 10 3 Baird-Parker 289 22 8 0 2.2 10 3 CODE-5 207 23 0 0 2.3 10 3 289 29 0 0 2.9 10 3 Baird-Parker 300 50 2 0 5.0 10 3 CODE-6 300 53 0 0 5.3 10 3 300 52 1 0 5.2 10 3 Baird-Parker 300 43 7 0 4.3 10 3 CODE-7 300 47 0 0 4.7 10 3 300 53 7 0 5.3 10 3 Baird-Parker 170 16 0 0 1.7 10 3 CODE-8 202 20 0 0 2.0 10 3 180 16 1 0 1.8 10 3 Baird-Parker 300 41 9 0 4.1 10 3 CODE-9 300 43 0 0 4.3 10 3 300 42 5 0 4.2 10 3 Baird-Parker 0 0 0 0 10 CODE-10 0 0 0 0 10 0 0 0 0 10
7, : 2761, CODE 1~9, CODE 10, Baird-Parker Z 2; 10 CFU/g, 3.2 盲样中添加菌株的鉴定及分型, CODE 1~10, 4 ( 2 ),, CODE 1 10-2 4, RiboPrinter, 1, 8, 4, #2 #8 (S. warneri) #6 #7 (S. epidermidis) #4 #5 (S. aureus) #1 #3 (S. simulans) EcoR Ⅰ, rdna, EcoRⅠ 8, 1.1~48 kb rdna, 8 4, 3.0%~91.8%, 2 S. aureus(#4 #5) 91.8%,, 3,,, 2 (#1 #3) 74.6%; 2 S. epidermidis (#6 #7) 70.7%; 2 S. warneri (#2 #8) 62.5%,,,, 4 结论, Z 2, Baird-Parker,, Baird-Parker,, ;,,,,, Baird-Parker Baird-Parker,,,,,,, 1, 1, GB4789.10-2010 Baird-Parker 2 3, Baird-Parker 1 RiboPrinter Fig. 1 Dendrogram of RiboPrinter clusters and relationships of adding bacteria in the sample
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