[用于测定牛奶中氯霉素的固相免疫酶分析方法的优化]

[Optimization of a method of solid-phase immunoenzyme analysis for determination of chloramphenicol in milk].

作者信息

Kolosova A Iu, Samsonova Zh V, Egorov A M, Shevaleva S A, Orlova N G, Kiseleva T V, Khotimchenko S A, Tutel'ian V A

出版信息

Vopr Pitan. 1999;68(1):23-7.

PMID:10198960

Abstract

The method of solid-phase enzyme linked immunosorbent assay (ELISA) for quantitative detection of chloramphenicol (CAP) in milk was developed. Peculiarities of the adsorption on the microtitre plates surface of CAP-ovalbumin conjugate were investigated. Different conditions of competition stage of the analysis were studied. Conditions providing CAP monitoring in human blood serum in the clinical range were optimized. Matrix effect on the assay results was studied. The specificity of the analytical system was investigated and the reagents stability was examined. The method developed permits CAP concentration to be determined in human blood serum, diluted 1/100, in the linear range from 0.1 to 100 ng/ml. The assay is characterized by high sensitivity (0.08 ng/ml) and good reproducibility (CV < 10.8%), assay time is about 3 hours. The correlation coefficient with HPLC is 0.977.

摘要

建立了用于定量检测牛奶中氯霉素（CAP）的固相酶联免疫吸附测定（ELISA）方法。研究了CAP-卵清蛋白偶联物在微量滴定板表面的吸附特性。研究了分析竞争阶段的不同条件。优化了在临床范围内监测人血清中CAP的条件。研究了基质对测定结果的影响。研究了分析系统的特异性并检测了试剂稳定性。所建立的方法能够在稀释1/100的人血清中测定CAP浓度，线性范围为0.1至100 ng/ml。该测定法具有高灵敏度（0.08 ng/ml）和良好的重现性（CV < 10.8%），测定时间约为3小时。与高效液相色谱法的相关系数为0.977。