不同载体酶联免疫技术检测牛乳中磺胺类药物的残留

Development of ELISA Formats Using Different Carriers for Sulfonamides Detection in Milk

本试验以抗磺胺类药物单克隆抗体为基础,分别建立了以酶标板和金磁微粒为载体的间接竞争酶联免疫吸附测定法,用以检测全脂牛乳中磺胺类药物残留。结果表明,两种方法的检测限均低于0.5ng/mL,对磺胺噻唑、磺胺甲恶唑和磺胺氯哒嗪的交叉率高,与其他抗生素未发现交叉反应;两种方法受全脂牛乳基质的干扰均较小,磺胺噻唑加标回收率为87.0%～95.8%,变异系数为3.9%～9.0%。与传统酶标板相比,金磁微粒提高了牛乳中磺胺类药物检测的灵敏度和精密度,缩短了检测时间。

For detection of Sulfonamide residues in full-cream milk samples, two indirect competitive enzyme- linked immunosorbent assays（CiELISA） have been developed on the base of prepared anti-sulfonamide group- specific monoclonal antibody, which using microtiter plates（MP） and gold magnetic nanoparticles（GMNP） as the carrier respectively. The results show that the detection limits of the two CiELISAs were both lower than 0.5 ng/mL. For two formats, sulfathiazole, sulfamethoxazole and sulfachlorpyridazine had high cross-reactivity rate and other common non-sulfonamide antibiotics had no cross-reactivity. Both formats were subject to less interference from full-cream milk matrix. The recovery values for sulfathiazole in full-cream milk samples spiked ranged from 87.0% to 95.8% and coefficient of variation ranged from 3.9% to 9.0%. The use of GMNP have improved the detection sensitivity, accuracy, precision and reduced the detection time compared with traditional microtiter plates.