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[使用标记抗原的酶联免疫吸附测定法检测食品中的葡萄球菌肠毒素A、B和C]

[Determination of staphylococcal enterotoxins A, B and C in foods using ELISA with labeled antigen].

作者信息

Windemann H, Baumgartner E

出版信息

Zentralbl Bakteriol Mikrobiol Hyg B. 1985 Dec;181(3-5):320-44.

PMID:4096149
Abstract

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.

摘要

采用根据施蒂夫勒 - 罗森伯格和费伊(1978年)方法制备的标记抗原竞争性酶联免疫吸附测定法(聚苯乙烯球),对食品中的金黄色葡萄球菌肠毒素A、B和C进行分析,以检测是否能够测定出与肠毒素A和B最大限量(分别为1纳克/克食品和10纳克/克食品)相对应的肠毒素浓度。研究了各种食品成分对竞争性酶联免疫吸附测定法单步和两步变体中肠毒素定量测定的影响。一般来说,食品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。所有肠毒素受到的影响相同。然而,在同一类食品中,发现标记和未标记抗原的结合存在显著差异。食品成分的影响通常取决于测定变体;奶酪提取物在两步法中效果更好,面食提取物在单步酶联免疫吸附测定中效果更好。在一些弱阳性提取物中,无法排除假阳性结果。在仅涉及食物中毒的奶酪样本中,仅发现了A型肠毒素(高达30纳克/克)。添加的肠毒素(1 - 10纳克/克食品)在奶酪中的回收率为50 - 70%,在面食食品中约为70%。缓冲液中的测定灵敏度对于肠毒素A和B为0.2纳克/毫升,对于肠毒素C为0.3纳克/毫升(20微升样本),对于A和B为0.5纳克/毫升且对于C为0.6纳克/毫升(5微升样本),对于A和B为1纳克/毫升且对于C为2纳克/毫升(1微升样本)。在食品中,尤其是在肉类和面食中,检测限通常更高。除了一些例外情况,仅在奶酪中能够达到肠毒素A所需的灵敏度(1纳克/克)。

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