Colloidal Gold Immunoassay Test Strip for Detecting Microcystin Antibody
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摘要: 用自制的微囊藻毒素(Microcystin-LR,MC-LR)单克隆抗体与钥孔血蓝蛋白(KLH)偶联物为胶体金标记底物研究MC-LR胶体金免疫检测技术。制备各种粒径的胶体金,最终确定粒径30nm的胶体金作为标记抗MC-LR单克隆抗体效果最佳;制备半抗原MC-LR与KLH的偶联物;在硝酸纤维素膜上包被MC-LR与KLH的偶联物作为检测带、包被羊抗鼠IgG作为质控带,建立快速检测MC-LR的免疫层析试纸条方法,该方法在3~5min即可目测判断结果,灵敏度为2ng·mL-1;依据工艺流程,发明制备了微囊藻毒素MC-LR的免疫层析快速检测卡,该卡检测样本中的微囊藻毒素MC-LR含量阈值为2ng·mL-1。Abstract: A colloidal gold immunoassay methodology was developed using the conjugate of the lab-made monoclonal antibody of diarrhetic shellfish poison,microcystin-LR(MC-LR),and KLH as the labeled substrate.The conjugate was produced following the immune competition law.Colloidal gold with various particle sizes were prepared.The particle size of 30 nm was selected for labeling the antibody.The rapid detection was carried out on an immunochromatographic strip made of a nitrocellulose membrane coated with the conjugate as the inspection band and the goat anti-mouse IgG as the quality control band. Without a need for additional equipment,this newly developed test strip could give a rapid,visual result in 3-5minutes with a sensitivity and a low threshold MC-LR content of 2ng·mL-1.
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Key words:
- microcystin /
- monoclonal antibody /
- colloidal gold /
- immunoassay
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