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副猪嗜血杆菌抗体间接ELISA检测方法的建立与应用

郑念广 谭实勇 何玲 夏芳 高娟 罗满林

中国畜牧兽医2011,Vol.38Issue(8):185-188,4.
中国畜牧兽医2011,Vol.38Issue(8):185-188,4.

副猪嗜血杆菌抗体间接ELISA检测方法的建立与应用

Establishment and Application of an Indirect ELISA for Detection of Antibody Against Haemophilus parasuis

郑念广 1谭实勇 2何玲 2夏芳 3高娟 1罗满林1

作者信息

  • 1. 华南农业大学,广东广州 510642
  • 2. 华农温氏畜牧股份有限公司,广东云浮527400
  • 3. 广东大华农动物保健品股份有限公司,广东云浮527400
  • 折叠

摘要

Abstract

An indirect ELISA coated with 121 ℃ high pressure Haemophilus parasuis serovar 4 and 5 hot-torlerance protein was established. The results of chessboard titration tests showed that coating antigen at a concentration of 10 μg/mL incubated optimally at 37 t for 2 h and then coated for 30 min with 20 g/mL skim milk powder. The optimal dilution of serum was 1 ∶ 80. The reaction time of coating antigen with serum 30 min. The optimal dilution of ELISA secondary antibody was 1∶ 12000. The reaction time was 30 min. The time of color development reaction was 15 min. Reproducibility,specificity,sensitivity tests and detection for 200 sera by the established ELISA confirmed that the developed ELISA had good stability and specificity. Compared with indirect haemagglutination test, the indirect ELISA was more sensitive and the clinical test result was consistent to overseas ELISA reagent kits. So this assay can be used as a tool for diagnosis and quarantine of H. parasuis.

关键词

副猪嗜血杆菌/抗体/间接酶联免疫吸附试验

Key words

Haemophilus parasuis/antibody/indirect ELISA

分类

农业科技

引用本文复制引用

郑念广,谭实勇,何玲,夏芳,高娟,罗满林..副猪嗜血杆菌抗体间接ELISA检测方法的建立与应用[J].中国畜牧兽医,2011,38(8):185-188,4.

基金项目

广东大华农动物保健品股份有限公司科研基金(033A201005-2). (033A201005-2)

中国畜牧兽医

OA北大核心CSTPCD

1671-7236

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