奶牛乳房炎乳中结合珠蛋白含量的夹心ELISA检测方法研究  

Determination of haptoglobin concentration in milk of cows with mastitis by sandwich enzyme-linked immunosorbent assay

作　　者：杨宇[1] 吕英[1] 石晶[1] 李庆章[1] 

机构地区：[1]东北农业大学乳品科学教育部重点实验室,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2011年第6期452-456,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十一五科技支撑计划重点项目;国家重点领域认证认可推进工程资助(2008BAK42B05)

摘　　要：为准确检测奶牛乳房炎乳中结合珠蛋白(Hp)含量,本研究采用牛血红蛋白(Hb)包被酶标板捕获乳清中Hp,利用特异性抗血清和酶标抗体建立夹心ELISA方法,并对建立的方法进行优化,用于检测牛奶中Hp含量。结果表明,ELISA方法检测下限为0.08μg/mL,与其他几种常见乳房炎乳中急性期蛋白质无交叉反应,批内试验和批间试验变异系数分别为3.27%～4.98%和5.46%～8.31%。应用本实验建立的夹心ELISA方法和商品化试剂盒分别检测50份已知患乳房炎牛乳清中Hp含量,同时对检测结果做相关性分析。结果表明,夹心ELSIA方法与商品化试剂盒具有很好的相关性(R2=0.996 2),同时验证了乳清中体细胞数与Hp含量呈正相关。夹心ELISA方法可准确测定牛奶中Hp含量,并可为奶牛乳房炎的诊断提供一种简易有效的辅助手段。To determinate the concentration of haptoglobin（Hp） in milk of cows with mastitis,a sandwich ELISA was established by coating with bovine-hemoglobin to capture Hp in whey and detecting with the anti-Hp-antibody and ap-conjugated-antibody.The Hp detection limit of ELISA was 0.08 μg/mL.The ELISA test had no cross-reaction with other acute phase proteins of cows with mastitis.The variant coefficients of ELISA intra-and inter-batches were from 3.27% to 4.98% and from 5.46% to 8.31%,respectively.Correlation between analytical results of the ELISA test and the commercial kits test was similar（R2=0.996 2） based on detection of 50 whey samples.This method could provide an simple and effective approach for the diagnosis of mastitis.

关 键 词：结合珠蛋白 牛血红蛋白 夹心ELISA 乳房炎 

分 类 号：S858.23[农业科学—临床兽医学]