磺胺二甲嘧啶的直接竞争酶联免疫吸附测定法。

Direct competitive enzyme-linked immunosorbent assay for sulfamethazine.

作者信息

Ko E, Song H, Park J H

机构信息

Departrment of Veterinary Medicine, Cheju National University, Korea.

出版信息

J Vet Med Sci. 2000 Oct;62(10):1121-3. doi: 10.1292/jvms.62.1121.

DOI:10.1292/jvms.62.1121

PMID:11073088

Abstract

A direct competitive enzyme-linked immunosorbent assay (ELISA) for screening sulfamethazine (SMZ) in pork tissues was developed. The assay was made with the affinity-purified polyclonal antibody-coated microtiter plate. A cross reactivity of IgG was observed at 3.5 microg/g of sulfamerazine among nine kinds of sulfonamide tested. Pork tissues fortified with SMZ was mixed with octadecyl silica (C18), and extracted with dichloromethane. The extracted SMZ was measured by homemade ELISA, commercial ELISA, and HPLC. The results were correlated (r=0.993, p<0.01). The homemade ELISA was sensitive to determine SMZ at the maximum residue level (MRL) as commercial one. During stability test of the IgG coated microtiter plate performed at 40 degrees C for 14 days, no difference in sensitivity was observed. We developed homemade ELISA with a detection limit of 10 ng of SMZ per g of pork tissues, and it could be used to screen SMZ in pork tissues.

摘要

建立了一种用于检测猪肉组织中磺胺二甲嘧啶（SMZ）的直接竞争酶联免疫吸附测定（ELISA）方法。该测定采用亲和纯化的多克隆抗体包被的微量滴定板。在九种测试的磺胺类药物中，观察到IgG在3.5微克/克的磺胺间二甲氧嘧啶处有交叉反应性。用十八烷基硅胶（C18）将添加了SMZ的猪肉组织混合，并用二氯甲烷萃取。萃取的SMZ通过自制ELISA、商用ELISA和高效液相色谱法（HPLC）进行测定。结果具有相关性（r = 0.993，p < 0.01）。自制ELISA在最大残留限量（MRL）下检测SMZ的灵敏度与商用ELISA相同。在40℃下对IgG包被的微量滴定板进行14天的稳定性测试期间，未观察到灵敏度差异。我们开发的自制ELISA检测限为每克猪肉组织10纳克SMZ，可用于筛查猪肉组织中的SMZ。