杂色曲霉素抗体的制备与应用  

Production and Application of Antibody against Sterigmatocystin

作　　者：楼建龙[1] 邹康南[1] 

机构地区：[1]南京农业大学兽医系,210014

出　　处：《中国公共卫生学报》1991年第6期367-369,共3页

摘　　要：本文提出了一种抗杂色曲霉素抗体的制备方法,建立了检测食物及饲料中杂色曲霉素(ST)污染情况的酶联免疫吸附试验(ELISA)和斑点酶联免疫吸附试验(Dot-ELISA)。两种方法对ST的最低检出限为2ppb(μg/kg)。液相萃取配合多次层析技术,从杂色曲霉纯培养物中提取到ST,经薄层层析和高效液相色谱鉴定后,以甲醛为偶联剂,将一定量ST连接到牛血清白蛋白(BSA)上,制得复合抗原BSA-ST,多次免疫大白鼠和青紫蓝兔,10～16周后获得不同滴度的抗体。用双向琼扩试验、对流免疫电泳和间接ELISA测定抗体的特异性和效价后,经饱和硫酸铵盐析和DEAE纤维素离子交换法纯化抗体。以ELISA和Dot-ELISA两种方法检测了24份饲料样品,发现其中14份含ST达8～6500ppb。Sterigmatocystin(ST) was extracted from a semi-synthetic corn-soybean culture of Aspergillus versicolor,incubated for 1 month and then conjugated to Bovine Serum Albumin (BSA) or Human Serum Albumin (HSA) by utilizing formaldehyde as linkage (Conjugon).ST Antibody was developed in rabbits and rats after reeated Finjection with BSA-ST conjugates.The specifity and titers of the antiserum were determined by an indirect Enzyme Linked Immuno Assay (ELISA)using HSA-ST conjugates as antigen.10～16 weeks after the first injection, the titers of the antibody range from 1:1600 to 1:32000. The 24 animal feedstuff samples were detected for ST contamination by a newly established Dot-ELISA method and an indirect competitive ELISA method,14 were found positive.The content was 8～6500 ppb (50ug/25ul).The detection limit for ST in animal feedstuff was 2 ppb (50/ug25ul)when the indirect competitive ELISA was applied.

关 键 词：杂色曲霉素 抗体 制备 ELISA 

分 类 号：R392－33[医药卫生—免疫学]