ELISA法测定食品中葡萄球菌肠毒素A、B和C

H Windemann, E Baumgartner
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引用次数: 0

摘要

采用stiff - rosenberg和Fey(1978)的标记抗原竞争性ELISA(聚苯乙烯球)对食品中的葡萄球菌肠毒素(SE) A、B和C进行分析,观察是否能测出SEA和SEB最高限量(分别为1 ng/g食品和10 ng/g食品)所对应的肠毒素浓度。研究了不同食品成分对竞争性酶联免疫吸附测定单步法和两步法中硒含量的影响。一般来说,食物的影响在奶酪提取物中最低,在肉类和面食产品提取物中最高。所有肠道毒素受到的影响相同。然而,在同一种食物中,标记抗原和未标记抗原的结合存在显著差异。食物成分的影响往往取决于测定变量;奶酪提取物在两步ELISA中效果更好,意大利面提取物在单步ELISA中效果更好。在一些弱阳性提取物中,不能排除假阳性结果。在涉及食物中毒的奶酪样品中,发现了A型肠毒素(高达30纳克/克)。添加SE (1-10 ng/g食物)的回收率在奶酪中为50-70%,在面食中约为70%。在缓冲液中的测定灵敏度范围从肠毒素A和B的0.2 ng/ml和肠毒素C的0.3 ng/ml (20 ml样品)到A和B的0.5 ng/ml和C的0.6 ng/ml (5 ml样品)到A和B的1 ng/ml和C的2 ng/ml (1 ml样品)。在食品中,特别是在肉类和面食中,检测限通常较高。除了一些例外,肠毒素A的灵敏度(1纳克/克)只能在奶酪中达到要求。
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[Determination of staphylococcal enterotoxins A, B and C in foods using ELISA with labeled antigen].

The competitive ELISA (polystyrene balls) with labeled antigen according to Stiffler-Rosenberg and Fey (1978) was applied to the analysis of Staphylococcal enterotoxins (SE) A, B and C in food to see if the enterotoxin concentration corresponding to maximum limit for SEA and SEB (1 ng/g food and 10 ng/g food respectively) could be measured. The effect of various food ingredients on the quantitative determination of SE in single-step and two-step variants of competitive ELISA was investigated. Generally, the effect of food was the lowest in extracts of cheeses and the highest in extracts of meats and pasta products. All enterotoxins were equally affected. However, within the same type of food significant differences in binding of both labeled and unlabeled antigen were found. The effect of food components often depended on the assay variant; the extracts of cheese gave better results in the two-step and the extracts of pasta better results in the single-step ELISA. In some weak-positive extracts, false positive results could not be excluded. In the samples of cheese which were involved in food poisoning exclusively the enterotoxin type A was found (up to 30 ng/g). The recovery of added SE (1-10 ng/g food) ranged from 50-70% in cheese and was about 70% in pasta foods. The assay sensitivity in buffer ranged from 0.2 ng/ml for enterotoxins A and B and 0.3 ng/ml for enterotoxin C (20 ml sample) to 0.5 ng/ml for A and B and 0.6 ng/ml for C (5 ml sample) to 1 ng/ml for A and B and 2 ng/ml for C (1 ml sample). In food, especially in meat and pasta the detection limit was often higher. With some exceptions the required sensitivity for enterotoxin A (1 ng/g) could only be reached in cheese.

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