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[抗伏马菌素B1单克隆抗体的研制及伏马菌素B1定量分析酶联免疫试剂盒]

[Development of monoclonal antibody against fumonisin B1 and ELISA-kit of quantitative analysis for fumonisin B1].

作者信息

Jiang Tao, Gong Hui-zhi, Li Feng-qin, Ji Rong

机构信息

Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100021, China.

出版信息

Wei Sheng Yan Jiu. 2006 Mar;35(2):209-12.

Abstract

OBJECTIVE

To detect the contamination level of fumonisin B1 in grain, and to develop rapid detection kit that possess patent of China.

METHODS

Hybridoma cell line excreting monoclonal antibody against fumonisin B, was produced using B cell hybridoma technique and develop a rapid, sensitive, quantitative ELISA-kit for detection fumonisin B1.

RESULTS

The monoclonal antibody used in the ELISA-kit were tested for subtype as IgG1 and its affinity constant was 8.3 x 10(-8) mol/L. The monoclonal antibody obtained in the present study was highly specific to fumonisin B1 , because no cross reactions between the monoclonal antibodies against fumonisin B1 with the analogues of fumonisin B, were found. The limited concentration of the ELISA-kit was 5 ng/ml, linear range was 50-500 ng/ml, the linear equation was Y = -0.582 x +1.793( = 0.99, P < 0.05). The recovery rate of maize on the level of 50n g/ml, 200 ng/ml and 500 ng/ml was among 71.89%-112.95%. The kit can be stored at normal tempertature in ten months at least. The coefficient of variant winthin-laboratory and between-laboratory was less than 20%.

摘要

目的

检测粮食中伏马菌素B1的污染水平,研制具有我国专利的快速检测试剂盒。

方法

采用B淋巴细胞杂交瘤技术制备分泌抗伏马菌素B单克隆抗体的杂交瘤细胞系,研制快速、灵敏、定量检测伏马菌素B1的ELISA试剂盒。

结果

该ELISA试剂盒所用单克隆抗体亚型鉴定为IgG1,亲和常数为8.3×10(-8)mol/L。本研究获得的单克隆抗体对伏马菌素B1具有高度特异性,因为未发现抗伏马菌素B1单克隆抗体与伏马菌素B类似物之间存在交叉反应。该ELISA试剂盒的检测限为5 ng/ml,线性范围为50 - 500 ng/ml,线性方程为Y = -0.582x +1.793(r = 0.99,P < 0.05)。玉米在50 ng/ml、200 ng/ml和500 ng/ml水平的回收率在71.89% - 112.95%之间。该试剂盒至少可在常温下保存十个月。实验室内和实验室间变异系数均小于20%。

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