猪表皮生长因子间接竞争ELISA检测方法的建立  

Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay of pEGF

作　　者：张万江[1] 王秀梅[1] 杨桂香[1] 

机构地区：[1]华南农业大学兽医学院广东省兽药研制与安全评价实验室,广东广州510642

出　　处：《动物医学进展》2008年第3期25-28,共4页Progress In Veterinary Medicine

基　　金：广州市科技攻关项目(2006Z3-E0051)

摘　　要：为建立酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测猪血清或尿液中猪表皮生长因子(porcinee pidermal growth factor,pEGF)的含量,以pEGF为包被原,人表皮生长因子(hu-mane pidermal growth factor,hEGF)为竞争抗原,两者与一定量的抗pEGF多抗反应。结果表明,理想的包被抗原浓度为0.625μg/mL,抗pEGF工作浓度为1∶64000,酶标二抗工作浓度为1∶20000,可检测的最适范围为0.4ng/mL^32ng/mL,最低检测限为1ng/mL,批内和批间变异系数分别为3.50%和5.22%。得到回归方程y=-34.53x+76.06(R2=0.993)和标准曲线,从而建立了快速定量测定pEGF含量的酶联免疫吸附试验方法。The aim of this paper was to establish the ELISA for detecting the concentration of porcine epidermal growth factor（pEGF） in porcine serum or urine, pEGF was used as coating antigen,and hEGF as competitive antigen. They were reacted to certain amount of polyclonal antibodies against pEGF. The result showed that the optimal concentration of the coating antigen, polyclonal antibodies against pEGF and sheep anti-rabbit IgG were 0. 625 μg/mL, 1 ： 64 000 and 1 ： 20 000, respectively. Quantization of the pEGF was linear from 0.4 ng/mL-32 ng/mL,with the detection limit being 0.4 ng/mL. The coefficients of variation of intra-assay and inter-assay were 3. 50% and 5. 22%, respectively. The regression equation was y= -34.53x＋76. 06（R^2 =0. 993） and there was a standard curve. Thus the indirect competitive ELISA for quantitative detecting pEGF was estabilished.

关 键 词：猪表皮生长因子 多克隆抗体 间接竞争ELISA 

分 类 号：S854.43[农业科学—临床兽医学]