8 1 Vol. 8 No. 1 2017 1 Journal of Food Safety and Quality Jan., 2017 2 * 王利刚, 张磊, 张婧, 费云滟, 付莎莉, 吴丹, 屠大伟 (, 401121) 摘要 : 目的 mini-vidas 2, 2 方法, 3, 3 结果 mini-vidas, mini-vidas, 3 结论 mini-vidas,,, mini-vidas 关键词 : ; mini-vidas ; ; Comparison of traditional culture method and 2 rapid screening methods for detection of Staphylococcus aureus WANG Li-Gang, ZHANG Lei, ZHANG Jing, FEI Yun-Yan, FU Sha-Li, WU Dan, TU Da-Wei * (Chongqing Institute for Food and Drug Control, Chongqing 401121, China) ABSTRACT: Objective To compare 2 rapid screening methods of mini-vidas method and membrane chip method with the culture method in national standard, evaluating the effectiveness and feasibility of the 2 methods. Methods Staphylococcus aureus was selected as target bacteria, the experiments were designed to test the sensitivity, specificity and interference resistance of 3 methods, and the artificially contaminated samples were tested simultaneously. The application effects of 3 screening methods were compared. Results The culture method in national standard and mini-vidas method had a high sensitivity, the specificity and interference resistance of mini-vidas and membrane chip method were better, and the results of 3 methods in artificially contaminate test were consistent with the design expectation. Conclusion The culture method in national standard and mini-vidas method can be used in daily inspections, and membrane chip method can be used in emergency testing in order to shorten the screening time. Additionally, the mini-vidas method and membrane chip method are reliable tools for rapid screening of Staphylococcus aureus. KEY WORDS: Staphylococcus aureus; mini-vidas method; membrane chip method; rapid screening 基金项目 : (cstc2015shmszx00002) Fund: Supported by the Social Science and Technology Innovation Project of Chongqing (cstc2015shmszx00002) * 通讯作者 :,, E-mail: weidatufu@163.com *Corresponding author: TU Da-Wei, Senior Engineer, Chongqing Institute for Food and Drug Control, Chongqing 401121, China. E-mail: weidatufu@163.com
1, : 2 239 1 引言 (Staphylococcus aureus),,,,,, [1-5],, [6], [7],, 36~72 h,,,,, [8-12] [7] [13] 3,, 2 材料与方法 2.1 材料 2.1.1 菌株与样品 (Staphylococcus aureus, CICC 23656) (Staphylococcus epidermidis, CICC 10294) ; (Salmonella typhimurium, ATCC 14028 ) (Listeria monocytogenes, ATCC 19115 ) (Enterobater sakazakii, ATCC 25944 ) (Escherichia coli, ATCC 25922 ), 50, 10 10 10 10 10 2.1.2 仪器与试剂 ZDP-2160 ( ); mini-vidas ( biomérieux, Inc.); MFS-8 ( ); IMG-1 ( ) 7.5%Baird-Parker BHI ( ); VIDAS SET2 ( biomérieux, SA ); DNA (()); () 2.2 实验方法 2.2.1 实验步骤 : GB 4789.10 [7], Baird-Parker, 36 24 h, mini-vidas :,, :,, DNA DNA, PCR Mix DNA, 99 10 min,, 2.2.2 灵敏性试验 3,, 36 24 h, 1:10, 10 1 ~10 4 CFU/mL,, 2.2.1 2.2.3 特异性试验 3,, 1:10, 10 3 CFU/mL, 2.2.1 2.2.4 杂菌干扰试验 3,, 1:10,, 3 1, 10 1 CFU/mL, 1:1:1:1:1 ; 2 5 10 2 CFU/mL, 1:1:1:1:1 ; 3 5 10 3 CFU/mL, 1:1:1:1:1 1:1(V:V) 10 2 CFU/mL, 10 3 CFU/mL 10 3 CFU/mL, 2.2.1 2.2.5 人工污染试验, 3, 50,
240 8 10 10 10 10 10 2,, 25 g 1 ml 10 2 CFU/mL ;,, 2.2.1 3 结果与分析 3.1 灵敏性实验 3 1,, Baird-Parker ( ) 10 1 CFU/mL; mini-vidas 10 1 CFU/mL,, (test value, TV) 0.13, ; 10 2 CFU/mL,, DNA PCR [14],,, mini-vidas 3.2 特异性实验 3 2, Baird-Parker, Baird-Parker : 2~3 mm,,,, [7],, Baird-Parker,,, Baird-Parker, [15] mini-vidas, spa [16],, mini-vidas 表 1 3 种检测方法的灵敏性比较 Table 1 Comparison of sensitivity of 3 methods (CFU/mL) 10 1 10 2 10 3 10 4 Mini-VIDAS + + + + TV 0.20 0.73 1.24 2.89 + + + + * - - + + : * 1 ; 5 ; 11 ; 12 表 2 3 种检测方法的特异性比较 Table 2 Comparison of specificity of 3 methods + - - - - - mini-vidas + - - - - - + - - - - -
1, : 2 241 3.3 杂菌干扰实验 3, 2.4, 10 1 CFU/mL 10 2 CFU/mL 10 3 CFU/mL,, 3 3, 3,, 3,,,,, [17] Mini-VIDAS,,,, DNA DNA, DNA, Table 3 表 3 3 种检测方法的抗干扰性比较 Comparison of anti-interference of 3 methods (CFU/mL) 10 1 10 2 10 3 +/- +/- +/- + +/- mini-vidas + + + + - + + - + - 3.4 人工污染实验 4, 90%, 10%,, mini-vidas 100%, 80%, 20%,,, [18], mini-vidas Table 4 表 4 人工污染样品的筛检结果比较 Comparison of the pre-detection for artificially contaminated samples (%) (%) + - +/- + - +/- 90 0 10 0 82 18 mini-vidas 100 0 / 20 80 / 100 0 / 0 100 /, 100%,,,,,, 4 结论 3,, ; mini-vidas,,, ;,,, 3, 36~72 h, mini-vidas 90 min 24 h, 14 h, mini-vidas,, mini-vidas, 参考文献 [1],,,. [J]., 2016, 35(2): 1 4. Zhou L, Wang Y, Wang FY, et al. Recent advance of general situation and novel detection technologies of Staphylococcus aureus in food [J]. China Brew, 2016, 35(2): 1 4. [2],,,. [J]., 2016, 7(2): 555 560. Zhang QQ, Fan YL, Song MH, et al. Research advances on foodborne Staphylococcal enterotoxins and its detection methods [J]. J Food Saf Qual, 2016, 7(2): 555 560. [3] Necidova L, Bogdanovicova K, Harustiakova D, et al. Short communication: Pasteurization as a means of inactivating staphylococcal enterotoxin A, B, and C in milk [J]. J Dairy Sci, 2016, 99(11): 8638 8643. [4] Cheng K, Wang S, Xu Y. Different roles of Staphylococcus aureus enterotoxin in different subtypes of nasal polyps [J]. Exp Therapeutic Med, 2017, (13): 321 326. [5]. [J]., 2004, 14(4): 392 395. Li Y. Research advances on foodborne staphylococcal enterotoxins and its detection methods [J]. Chin J Health Lab Technol, 2004, 14(4): 392 395. [6] GB 29921-2013 [S]. GB 29921-2013 National food safety standard Pathogen limits for food [S]. [7] GB 4789.10-2010 [S]. GB 4789.10-2010 National food safety standard Food microbiological examination: Staphylococcus aureus [S]. [8],,,. PCR [J]., 2016, 42(3): 198 201.
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