6 5 Vol. 6 No. 5 2015 5 Journal of Food Safety and Quality May., 2015 Mini-VIDAS 马 * 晨 (,, 571101) 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 571101, 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 : (20153163) (1630052014010) Fund: Supported by the Natural Science Foundation of Hainan Province (20153163) and the Fundamental Scientific Research Funds for Chinese Academy of Tropical Agricultural Sciences (1630052014010) * 通讯作者 :,, E-mail: mc19860112@163.com *Corresponding author: MA Chen, Assistant Research Fellow, Analysis and Testing Center Chinese Academy of Tropical Agriculture Science, Haikou 571101, China. E-mail: mc19860112@163.com
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 2.1.1 菌株及试剂 (Listeria monocytogenes CICC 21633/ATCC 19111) (CICC 21671) (CICC 21663/ATCC19119) (CICC 10417/ATCC 33090) mini-vidas II(LMO2) ( ) ( ) 2.1.2 样品基质 ; ( ; ),,,
1852 6 2.2 试验方法 2.2.1 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 ( 2.0~2.8 10 8 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] 2.2.2 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] 2.2.3 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, 10 4-10 5 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 + - - - - : -, ; +, ; +/-
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 10 3 10 4 10 5 10 6 - +/- + + 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 1/10 6 + + + + - - 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, 1 0.5 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 ( 10 0-10 1 cfu/ml) LB 2 FB 2,,, mini-vidas LB 2 FB 2,, 1.1 10 7 cfu/ml 4.0 10 6 cfu/ml 8.1 10 6 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 + + + + - - + + + + : -, ; +,
1854 6 1 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 4~6 10 5 5~7 10 4 3~5 10 4 4~6 10 3 <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 2 + - - + - - - ++ FB 2 + - - - - - - ++ Fraser + - - + + + - ++ mini-vidas LB 2 + - - + - - - ++ FB 2 + - - - - - - ++ Fraser + - - + + + - ++
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