食品科学

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间接竞争ELISA法检测牛乳中β-乳球蛋白含量的准确性评价

袁水林1,2,熊 鼎1,2,陈红兵1,3,高金燕2,李 欣1,2,*   

  1. 1.南昌大学 食品科学与技术国家重点实验室,江西 南昌 330047;2.南昌大学生命科学与食品工程学院,江西 南昌 330047;
    3.南昌大学中德联合研究院,江西 南昌 330047
  • 出版日期:2014-09-25 发布日期:2014-09-17
  • 通讯作者: 李 欣
  • 基金资助:

    “十二五”国家科技支撑计划项目(2012BAK17B02);国家自然科学基金青年科学基金项目(31260204;31301522);
    江西省自然科学基金项目(2012BAB204002);江西省教育厅课题(GJJ12143)

Evaluation of the Accuracy of Indirect Competitive ELISA Used to Detect Milk β-Lactoglobulin

YUAN Shui-lin1,2, XIONG Ding1,2, CHEN Hong-bing1,3, GAO Jin-yan2, LI Xin1,2,*   

  1. 1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
    2. School of Life Sciences and Food Engineering, Nanchang University, Nanchang 330047, China;
    3. Sino-German Joint Research Institute, Nanchang University, Nanchang 330047, China
  • Online:2014-09-25 Published:2014-09-17
  • Contact: LI Xin

摘要:

研究建立了以β-乳球蛋白标准品为包被抗原、自制抗体为一抗、辣根过氧化物酶标记的羊抗兔IgG∶HRP为二抗、邻苯二胺为底物的间接竞争酶联免疫检测法(indirect competitive enzyme-linked immunosorbent,ELISA)。同时以反相高效液相色谱法定量检测相同样品中β-乳球蛋白含量,采用t检验方法来评价间接竞争ELISA的方法定量检测β-乳球蛋白的准确性。t检验方法结果显示:在显著水平α=0.05下,该间接竞争ELISA法检测β-乳球蛋白的测量值较为准确。

关键词: 牛乳过敏, &beta, -乳球蛋白, 间接竞争ELISA法, 反相高效液相色谱

Abstract:

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was established with the standard
β-lactoglobulin as the coated antigen, specific rabbit antibody against β-lactoglobulin as the primary antibody, horseradish
peroxidase labeled goat anti-rabbit IgG:HRP as the secondary antibody and o-phenylenediamine as the substrate solution.
Reversed-phase high-performance liquid chromatography (RP-HPLC) was used as the reference method. The t-test method
proved that the indirect competitive ELISA for detection of β-lactoglobulin was feasible and accurate. The results of t-test
at a significance level α = 0.5 showed that the indirect competitive ELISA was more accurate to detect the concentration of
β-lactoglobulin.

Key words: milk allergy, β-lactoglobulin, indirect competitive enzyme-linked immunosorbent assay, reversed-phase highperformanceliquid chromatography

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