摘要
通过合成氯霉素完全抗原,制备氯霉素的多克隆人工抗体,建立氯霉素(CAP)的免疫学检测方法。采用重氮法合成氯霉素完全抗原,以氯霉素牛血清白蛋白(CAP-BSA)免疫大耳白家兔,用ELISA方法进行鉴定和特性分析。结果显示,该抗血清与链霉素、青霉素、四环素等常见抗生素的交叉反应率均小于2.1%,IC50为13.65 ng/ml,检测限达到1.01μg/L。获得了选择性好、特异性较高的氯霉素特异抗体,可用于氯霉素的快速定量检测。
Chloramphenicol（ CAP） is a inexpensive and effective broad-spectrum antibiotic that has been used in veterinary practice to treat septicaemia,pulmonary,urinary and digestive infections. However,two types of CAP-induced toxicity in humans have been identified,due to its side effects in humans,especially fatal bone marrow depression and aplastic anemia. The use of CAP for treatment of food-producing animals is prohibited in many countries including China.The detection limit of the analytical methods is crucial and decisive to control CAP residues at trace levels. Therefore,it is necessary to develop sensitive methods for determining CAP residues in animal tissues. But traditional detecting methods such as gas chromatography（ GC）,liquid chromatography（ LC）,GC-mass spectrometry（ MS） or LC-MS-MS been described in recent years are time consuming and require extensive clean-up steps,sophisticated equipment and well-trained personnel. Thus,there is a pressing need for the development of a rapid,sensitive and economic alternative screening method to detect the CAP residues. In this study,diazotization reaction were performed to synthesize the immunogen and coating antigen of chloramphenicol. White rabbits were immunized with BSA-CAP-HS,and the characteristics of polyclonal antibody（ p Ab） was identified by indirect enzyme-linked immunosorbent assay（ ELISA）. ELISA results showed that the cross-reactivities of the anti-serum with streptomycin,penicillin,tetracycline,were less than 2. 1%,and the hemi-inhibitory concentration（ IC50） was 13. 65 ng/ml. Recovery test revealed that the detection limit for CAP was 1. 01 ng/ml. The polyclonal antibody achieved in this study can be used for the sensitive and quantitative analysis of chloramphenicol.
出处
《江苏农业学报》
CSCD
北大核心
2015年第1期192-196,共5页
Jiangsu Journal of Agricultural Sciences
基金
浙江省科技计划项目(2013C37045)
关键词
氯霉素
多克隆抗体
酶联免疫吸附法
chloramphenicol
polyclonal antibody
enzyme-linked immunosorbent assay（ELISA）
