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用于快速筛查猪肌肉组织中氯霉素残留的灵敏链霉亲和素-生物素酶联免疫吸附测定法。

Sensitive streptavidin-biotin enzyme-linked immunosorbent assay for rapid screening of chloramphenicol residues in swine muscle tissue.

作者信息

van de Water C, Haagsma N

机构信息

University of Utrecht, Faculty of Veterinary Medicine, Department of the Science of Food of Animal Origin, The Netherlands.

出版信息

J Assoc Off Anal Chem. 1990 Jul-Aug;73(4):534-40.

PMID:2211475
Abstract

A sensitive, competitive enzyme-linked immunosorbent assay (ELISA) for chloramphenicol (CAP) in swine muscle tissue has been developed. The ELISA is based on an earlier procedure. To improve sensitivity, different optimization procedures were investigated. The introduction of a streptavidin-biotin system and the use of a coating antigen with a lower CAP incorporation resulted in the most sensitive ELISA: the CAP concentration giving 50% inhibition decreased from 125 ng/mL to 3.0 ng/mL. This ELISA procedure was applied for a rapid screening of CAP residues in swine muscle tissue. The tissues were extracted with demineralized water. A concentrated phosphate-buffered saline solution was added to the filtered aqueous extract and this sample solution was directly submitted to the ELISA procedure. The results were compared to values obtained by analysis of a corresponding blank. This blank was prepared by treating a part of the aqueous sample solution with an immobilized monoclonal antibody preparation. This treatment was necessary because aqueous extracts of different swine muscle tissues showed a high variation in dose-response curves, probably caused by the complexity and variability of the matrix. In spiked tissues, the presence of CAP at concentrations of 10 micrograms/kg and higher can be easily demonstrated.

摘要

已开发出一种用于检测猪肌肉组织中氯霉素(CAP)的灵敏、竞争性酶联免疫吸附测定(ELISA)方法。该ELISA基于早期方法。为提高灵敏度,研究了不同的优化程序。引入链霉亲和素-生物素系统以及使用氯霉素掺入量较低的包被抗原,得到了最灵敏的ELISA:产生50%抑制率的氯霉素浓度从125 ng/mL降至3.0 ng/mL。该ELISA程序用于快速筛查猪肌肉组织中的氯霉素残留。组织用去离子水提取。向过滤后的水提取物中加入浓缩磷酸盐缓冲盐溶液,然后将该样品溶液直接进行ELISA程序。将结果与通过分析相应空白获得的值进行比较。该空白通过用固定化单克隆抗体制剂处理一部分水样来制备。这种处理是必要的,因为不同猪肌肉组织的水提取物在剂量-反应曲线上表现出很大差异,这可能是由基质的复杂性和变异性导致的。在加标组织中,浓度为10微克/千克及以上的氯霉素的存在很容易被检测到。

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