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[使用标记抗体夹心酶联免疫吸附测定法测定食品中的葡萄球菌肠毒素A、B、C和D]

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

作者信息

Windemann H, Baumgartner E

出版信息

Zentralbl Bakteriol Mikrobiol Hyg B. 1985 Dec;181(3-5):345-63.

PMID:4096150
Abstract

The noncompetitive Sandwich-ELISA (polystyrene balls) with labelled antibody for staphylococcal enterotoxins (SE) A, B, C, and D according to Fey et al. (1984), which have recently been introduced commercially, was applied to analysis of foods. The effect of various food ingredients on the quantitative determination and recovery of SE was investigated. The unspecific effect of food components which, in some cases, caused false-positive results in the competitive ELISA was less frequent. Nevertheless, the differences in binding of unlabeled antigen between buffer and food were still observed. Using ELISA microtiter plates the effect of food components was less pronounced. Enterotoxin type A or a mixture of A and D were dominant in foods which were involved in food poisoning (about 150 samples). In meat the recovery of SE added (1-10 ng/g) ranged from 30-60%. The assay sensitivity in buffer ranged from 0.1 ng/ml for enterotoxins A, B, C and D (5 ml sample) to 0.2 ng/ml for enterotoxins A-D (1 ml sample) using polystyrene balls and was 1 ng/ml for enterotoxins A-D using ELISA plates. In foods the detection limit was occasionally higher. The required sensitivity for enterotoxin A (maximum limit 1 ng/g) could mostly be reached. Because of the differences in antigen binding in buffer and in various food extracts the determination of the enterotoxin content in this range is laborious.

摘要

根据Fey等人(1984年)的方法,采用带有标记抗体的非竞争性夹心酶联免疫吸附测定法(聚苯乙烯球)检测葡萄球菌肠毒素(SE)A、B、C和D,该方法最近已商业化推出,并应用于食品分析。研究了各种食品成分对SE定量测定和回收率的影响。食品成分的非特异性影响在某些情况下会在竞争性酶联免疫吸附测定中导致假阳性结果,这种情况较少见。然而,仍观察到缓冲液和食品中未标记抗原结合的差异。使用酶联免疫吸附测定微量滴定板时,食品成分的影响不太明显。在涉及食物中毒的食品(约150个样品)中,A型肠毒素或A和D型混合物占主导地位。在肉类中,添加的SE(1 - 10 ng/g)回收率在30% - 60%之间。使用聚苯乙烯球时,缓冲液中该测定方法的灵敏度范围为:A、B、C和D型肠毒素为0.1 ng/ml(5 ml样品),A - D型肠毒素为0.2 ng/ml(1 ml样品);使用酶联免疫吸附测定板时,A - D型肠毒素为1 ng/ml。在食品中,检测限偶尔会更高。对于A型肠毒素所需的灵敏度(最大限量1 ng/g)大多能够达到。由于缓冲液和各种食品提取物中抗原结合存在差异,在此范围内测定肠毒素含量较为费力。

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