Microsoft Word 马晨-二校出1850、1851、1853、1854请不要动版

Size: px

Start display at page:

Download "Microsoft Word 马晨-二校出1850、1851、1853、1854请不要动版"

椅笞云
7 years ago
Views:

1 6 5 Vol. 6 No Journal of Food Safety and Quality May., 2015 Mini-VIDAS 马 * 晨 (,, ) 摘要 : 目的, 方法 (CICC 21633/ATCC 19111) (CICC 21671) (CICC 21663/ATCC19119) (CICC 10417/ATCC 33090), mini-vidas 结果, mini-vidas 10 4 ~10 5 cfu/ml 10 5 cfu/ml;,, 1/10 4 < < < <, Fraser >LB 2 >FB 2 结论 mini-vidas, mini-vidas 关键词 : ; mini-vidas; ; Performance comparison of mini-vidas and Chinese national standard method for detection of Listeria monocytogenes MA Chen * (Hainan Provincial Key Laboratory of Quality and Safety for Tropical Fruits and Vegetables, Analysis and Testing Center Chinese Academy of Tropical Agriculture Science, Haikou , China) ABSTRACT: Objective The examination effect of Listeria monocytogenes with two methods were compared in the aspects of specificity, sensitivity and anti-interference. Effects of additives and sample matrix on the detection results were also studied. Methods Listeria monocytogenes (CICC 21633/ATCC 19111), L.seeligeri (CICC 21671), L.ivanovii (CICC 21663/ATCC19119) and L. innocua (CICC 10417/ATCC 33090) were used. Chinese national standard (GB) and mini-vidas method were utilized to test the Listeria monocytogenes in samples. Results The methods of mini-vidas and GB both had high specificity. The sensitivity was 10 4 ~10 5 cfu/ml for GB method and 10 5 cfu/ml for mini-vidas. GB method had a higher anti-interference than mini-vidas method. Among the tested additives, only sodium hypochlorite had an 基金项目 : ( ) ( ) Fund: Supported by the Natural Science Foundation of Hainan Province ( ) and the Fundamental Scientific Research Funds for Chinese Academy of Tropical Agricultural Sciences ( ) * 通讯作者 :,, mc @163.com *Corresponding author: MA Chen, Assistant Research Fellow, Analysis and Testing Center Chinese Academy of Tropical Agriculture Science, Haikou , China. mc @163.com

2 5 : Mini-VIDAS 1851 impact on the detection results. Inhibition percent of addictives on L. monocytogenes growth followed the order: ethanol<sodium benzoate<traditional spicy condiment (Shisanxiang)<vegetable salad condiment<sodium hypochlorite. When there were high bacterial background and small infective dose of L. monocytogenes in sample matrix, the detection results were interfered intensively. Effect of 3 enrichment medium for L. monocytogenes followed the order: Fraser broth>lb 2 >FB 2. Conclusion The detective characteristics of mini-vidas and GB methods are relatively similar. When using the GB method to detect L. monocytogenes, the characteristics of sample matrix should be paid more attention on, such as bacterial number and food additives. Additionally, mini-vidas is a good tool for pre-detection of the Listeria monocytogenes. KEY WORDS: Listeria monocytogenes; mini-vidas; addictives; sample matrix 1 引言 (Listeria monocytogenes), ( ) [1,2], 20%~30% [3],, 4, [4,5], [6,7], 2000 WHO [8],, [4], mini-vidas, L. monocytogenes,, [7],,,,,,,,,,, [9,10] VIDAS L. monocytogenes,, mini-vidas,,, mini-vidas L. monocytogenes,,, L. monocytogenes, / L. monocytogenes [4], mini-vidas,, ( ) ( ), 2 材料与方法 2.1 材料菌株及试剂 (Listeria monocytogenes CICC 21633/ATCC 19111) (CICC 21671) (CICC 21663/ATCC19119) (CICC 10417/ATCC 33090) mini-vidas II(LMO2) ( ) ( ) 样品基质 ; ( ; ),,,

3 试验方法特异性灵敏度和抗干扰性检测 5, 30, 10 6 cfu/ml,, [9,11], (BHI)30, 18~24 h, 2.5~3.0 ( 2.0~ cfu/ml), (10 0 ~10 7 cfu/ml), [9,11],, 1/10 1/0 2 1/10 3 1/10 4 1/10 5 1/10 6 /,, [12,13] 添加成分对检测效果的影响, 10 5 cfu/ml 10 6 cfu/ml, 0.5 g/l 1.0 g/l 75%,, mini-vidas, : A(%)=[(T 0 T 1 )/T 0 ] 100% T 0 ; T 1 ; [9,11] 样品基质对检测效果的影响 10 g ( 10 ml ), 0.6 ml 10 0 ~10 3 cfu/ml, LB 1 / LB [14] 2, Half-Fraser /FB 2 Half-Fraser/Fraser, mini-vidas 3 结果与讨论 3.1 方法特异性灵敏度和抗干扰性检测 ( ), 1 mini-vidas,,, ( ),,,, [15], VITEK, VIDAS-VITEK [16],, 2 PALCAM PALCAM, 10 4 cfu/ml,, 10 5 cfu/ml, cfu/ml, ;, [17] mini-vidas 10 5 cfu/ml, 3, mini-vidas 1/10 4,, mini-vidas Table 1 表 1 不同检测方法特异性 Comparison of specificity of two methods + +/- +/- - - mini-vidas : -, ; +, ; +/-

4 5 : Mini-VIDAS 1853 Table 2 表 2 不同检测方法灵敏度比较 Comparison of sensitivity of two methods /- + + mini-vidas : -, ; +, Table 3 表 3 不同检测方法抗干扰性比较 Comparison of anti-interference of two methods / 1/10 1/10 2 1/10 3 1/10 4 1/10 5 1/ mini-vi DAS : -, ; +, 3.2 添加成分对单增李斯特菌检出的影响 0.5 g/l 1.0 g/l 75%,, 4,, mini-vidas,, mini-vidas, g/l (0.1%)< (6.8%)< (26.5%)< (46.0%)< (98.3%) 1.0 g/l (0.5%)< (35.7%)< (37.0%)< (45.6%)< (100%), < < < < (P<0.01),,, 3.3 样品基质对单增李斯特菌检出的影响, 5, > > > > > > PALCAM, 6 ( cfu/ml) LB 2 FB 2,,, mini-vidas LB 2 FB 2,, cfu/ml cfu/ml cfu/ml, 10 4 cfu/ml, 6,,,,, Half-Fraser Fraser, mini-vidas ( 6),, Fraser LB 2 FB 2 Table 4 表 4 不同添加成分对检测结果的影响 Effects of different additions on the detection of Listeria monocytogenes 0.5 g/l 1.0 g/l 0.5 g/l 1.0 g/l 0.5 g/l 1.0 g/l 0.5 g/l 1.0 g/l 0.5 g/l 1.0 g/l mini-vidas : -, ; +,

5 Fig. 1 Inhibition of addictives on Listeria monocytogenes growth 表 5 不同食品基质菌落总数 Table 5 Total bacterial number in matrix (cfu/g) >10 8 4~ ~ ~ ~ <10 3 <10 2 (10 2 ~10 3 cfu/ml), ( ),, 10 2 cfu/ml 7 > > > > Fraser >LB 2 >FB 2, Fraser >LB 2 >FB 2 [11], (10 1 cfu/ml), mini-vidas ; (10 2 cfu/ml), 4 结论 :, mini-vidas, VITEK 10 4 ~10 5 cfu/ml mini-vidas 10 5 cfu/ml,, 1/10 4,, mini-vidas, mini-vidas,, 表 6 人工污染的基质中检出结果 ( 低添加浓度 10 0 ~10 1 cfu/ml) Table 6 Detection results of Listeria monocytogenes in artificial contaminated samples (added low concentration 10 0 ~10 1 cfu/ml) LB FB Fraser mini-vidas LB FB Fraser

6 5 : Mini-VIDAS 1855 表 7 增菌后基质中单增李斯特菌数量 ( 高添加浓度 10 2 cfu/ml) Table 7 Number of Listeria monocytogenes in matrix after selective enrichment (added high concentration 10 2 cfu/ml) ( : 10 7 CFU/mL) LB FB Fraser : (10 5 cfu/ml 10 6 cfu/ml), mini-vidas, < < < <,, :, ;,, Fraser >LB 2 >FB 2,, mini-vidas, mini-vidas,,,,,,,, 参考文献 [1] Todd ECD, Notermans S. Surveillance of listeriosis and its causative pathogen, Listeria monocytogenes [J]. Food Control, 2011, 22(9): [2] Meyer C, Fredriksson-Ahomaa M, Kleta S, et al. Occurrence of L. monocytogenes in ready-to-eat poultry products available on the German market [J]. Food Res Int, 2012, 48(2): [3] Lukinmaa S, Miettinen M, Nakari UM, et al. Listeria monocytogenes isolates from invasive infections: variation of sero- and genotypes during an 11-year period in Finland [J]. J Clin Microbiol, 2003, 41(4): [4],,,. [J]., 2013, 32(11): Zhou Y, Zhu CQ, Guo YC, et al. Comparasive study of nucleic acid extraction method of L.monocytogenes in the different kinds of food matrix [J]. J Food Sci Biotechnol, 2013, 32(11): [5],,,. PMA-qPCR [J]., 2014, 35(1): Huang Y, Yu YG, Huang XL, et al. Induction and PMA-qPCR detection of Listeria monocytogenes in viable but non-culturable state [J]. Sci Technol Food Ind, 2014, 35(1): [6] Chen J, Zhang X, Mei L, et al. Prevalence of Listeria in Chinese food products from 13 provinces between 2000 and 2007 and virulence characterization of Listeria monocytogenes isolates [J]. Foodborne Pathog Dis, 2009, 6(1): [7],. PCR mini-vidas [J]., 2010, 20(7): Lv J, Zheng HY. Combination use of PCR and mini-vidas in rapid detection of food for Listeria monocytogenes isolates [J], Chin J Health Lab Technol, 2010, 20(7): [8] Ryser T, Elmer HM. Listeriosis and food safety[m]. Marcel Dekke Inc, 1991, 6(2): [9],,,. VIDAS VITEK [J]., 2008, 33(9): Lin T, Lan M, Huang W, et al. Detection and classification of Listeria species in foods by VIDAS-VITEK [J]. Food Sci

7 Technol, 2008, 33(9): [10],. VIDAS [J]., 2015, 6(2): Wang SJ, Gao C. Fast detection of Salmonella in health food by VIDAS method [J]. J Food Saf Qual, 2015, 6(2): [11]. [D]. :, Lin SJ. The study of enzyme-linked fluorescence immunoassay for three food-borne pathogens [D]. Hangzhou: Zhejiang University of Technology, [12],,,. Mini-VIDAS PCR [C]., 2005: Liu PH, Wang J, Zhang SH, et al. Application of mini-vidas and PCR to detect Salmonella in food of animal source[c]. the Sixth National Assembly and Symposium of Chinese Association Of Animal Science and Veterinary Medicine. 2005: GB National food safety standard food microbiological examination: Listeria monocytogenes [S] [15],,,. [J]., 2005, 6(5): Chen HY, Hong CJ, Li Y, et al. The detection of Listeria in food and results of drug sensitivity test [J]. Chin Trop Med, 2005, 6(5): [16],,,. - [J]., 2008, 34(8): Lin T, Lan M, Huang W, et al. Application on VIDAS-VITEK to detect Salmonella spp. in auatic products [J]. Food Ferment Ind, 2008, 34(8): [17],,,. 5 [J]., 2011, 36(6): Fan Y, Wang SY, Chen NN, et al. Performance comparison of common culture media for isolation of Salmonella [J]. Food Sci Technol, 2011, 36(6): [13],,,. [J]., 2014, 35(15): Huang BY, Yu ZY, Lin YW, et al. Comparison of detection of Salmonella in food by four methods [J]. Sci Technol Food Ind, 2014, 35(15): [14] GB [S] 作者简介 ( 责任编辑 : 白洪健 ) 马晨, 助理研究员, 主要研究方向为食品和环境微生物 mc @163.com

Microsoft Word 聂雪梅.doc

Microsoft Word 聂雪梅.doc 4 4 Vol. 4 No. 4 2013 8 Journal of Food Safety and Quality Aug., 2013 聂雪梅 *, 李立, 孙利, 高飞 (, 100123) 摘要 : 2011 12,, 2007~2011,,,, 关键词 : ; ; ; ; Trade and notification situation on food import and export