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Position: Home > Articles > Establishment of immunological detection methods of PMSG Heilongjiang Animal Science and Veterinary Medicine 2019 (17) 12-15

PMSG免疫学检测方法的建立

作  者：
于艳昌;严慧;李闰婷;张英杰;齐伟民;段春辉;刘月琴
单  位：
河北农业大学动物科技学院
关键词：
孕马血清促性腺激素(PMSG);马属动物;早孕检测;抗体;ELISA;吸光度值
摘  要：
为了建立一种马属动物孕马血清促性腺激素(pregnant mare serum gonadotropin,PMSG)浓度的检测方法,试验使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)检测商用PMSG抗原纯度,斑点印迹杂交(Dot-blot)判断PMSG和抗体是否结合,蛋白免疫印迹杂交(Western-blot)确定抗体和PMSG抗原的结合方式,应用酶联免疫吸附试验(ELISA)建立PMSG浓度-吸光度值标准曲线,并拟合方程。结果表明:PMSG抗原纯度符合要求,抗体与PMSG抗原采用空间表位方式结合;PMSG浓度与OD_(450)值的标准曲线拟合方程为y=0.004 5x+0.020 2,R~2=0.998,在0～4×10~2IU/m L范围内,PMSG浓度与OD_(450)值之间高度相关,R~2>0.99。说明PMSG抗原可与抗体通过空间表位方式特异性结合,所建立的PMSG免疫学检测方法具有较高的准确性。
译  名：
Establishment of immunological detection methods of PMSG
作  者：
YU Yanchang;YAN Hui;LI Runting;ZHANG Yingjie;QI Weimin;DUAN Chunhui;LIU Yueqin;College of Animal Science and Technology,Hebei Agricultural University;Fuping County Animal Husbandry Bureau;
关键词：
pregnant mare serum gonadotropin(PMSG);;equine animals;;early pregnant diagnosis;;antibody;;ELISA;;absorbancy
摘  要：
In order to establish a method for detecting the concentration of pregnant mare serum gonadotropin( PMSG) in equine animals,the sodium dodecyl sulfate polyacrylamide gel electrophoresis( SDS-PAGE) was used in this experiment to detect the purity of commercial PMSG antigen; Dot-blot hybridization( Dot-blot) was used to determine whether PMSG and antibody were combined,protein immuno-imprinting hybridization( Western-blot) was used to determine the binding mode of antibody and antigen,and enzyme linked immunosorbent assay( ELISA) was used to establish the standard curve of PMSG concentration-absorbanceand the equation was fitted. The results showed that the purity of PMSG antigen was up to the requirement,the antibody binded to PMSG antigen in a spatial epitope mode,and the standard curve fitting equation of PMSG concentration and OD450 value was y = 0. 004 5 x+0. 020 2,R2= 0. 998.The results suggested that PMSG antigen could specifically bind to antibody through spatial epitope mode.PMSG concentration was highly correlated with OD450 value in the range of 0-4× 102 IU/m L,R2>0. 99. PMSG detection method had high accuracy.

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