应用GPV VP3基因重组原核表达产物建立检测抗体的ELISA方法研究  

Development of ELISA Methods to Detect Antibody by Prokaryotic Recombinant Expression Product of GPV VP3 Gene

作　　者：布日额[1] 李宝臣[2] 马波[1] 王君伟[1] Ulrich Neumann 

机构地区：[1]东北农业大学动物医学院 [2]农业部青岛动植物检疫局,青岛266000 [3]汉诺威兽医学院,汉诺威30559

出　　处：《畜牧兽医学报》2006年第2期199-203,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：黑龙江省教育厅科学技术研究项目(重大项目10541z004)

摘　　要：利用鹅细小病毒（GPV）VP3基因的原核表达蛋白作为包被抗原建立了检测GPV抗体的间接ELISA及Dot-ELISA方法。经确定两种方法的抗原包被浓度为125μg／mL，其中间接-ELISA 100μL／孔、Dot-ELISA 5μL／点。间接-ELISA中HRP标记的兔抗鹅IgG的工作浓度是1：200，检测血清的最适稀释度是1：400，阳性判定标准为OD492≥0．20，且P／N≥2．0。用此方法检测弱毒疫苗免疫血清，其抗体滴度在1：400～1：51200。Dot-ELISA的结果与间接-ELISA的结果一致。Indirect-enzyme-linked immunosorbent assay （Indirect-ELISA） and nitrocellulose blotting membrane （dot blot ELISA） were developed for the detection of antibodies against GPV in goose by recombinant GPV VP3 gene prokaryotic expression product. The amount of incubating antigen were 12.5 μg and 0. 625μg respectively. And working concentration of rabbit anti-goose IgG-HRP antibody was 1 ： 200, the suitable concentration of detected sera was 1 ＂ 400. The standards to estimate positive were OD492 ≥0.2, P/N ≥2.0 for the Indirect-ELISA method. ELISA antibody titers of detected sera immunized with attenuated vaccine were in 1 ： 400 1 ： 51 200. The results of Dot ELISA were consistent with the results of Indirect-ELISA.

关 键 词：鹅细小病毒 VP3基因重组原核表达产物 间接-ELISA Dot—ELISA 

分 类 号：S854.43[农业科学—临床兽医学]