摘要
用胸膜肺炎放线杆菌血清2型(1536)菌液与弗氏不完全佐剂等体积混合乳化后免疫大白兔,制备兔高免血清,提纯抗体,建立检测猪传染性胸膜肺炎血清2型抗体的双夹心ELISA方法。方阵滴定确定最佳反应条件:包被纯化的兔抗App血清2型抗体为9.375μg/mL,抗原稀释度为1∶1 600,HRP-兔抗猪IgG稀释度为1∶20 000。特异性与敏感性试验结果表明,双夹心ELISA方法特异性好、敏感性高,与猪O型口蹄疫、猪传染性胃肠炎、蓝耳病等阳性血清以及App其他血清型阳性血清无交叉反应。检测样品结果与阻断ELISA比较,其相符率为99.54%,表明双夹心ELISA可用于猪传染性胸膜肺炎血清2型抗体的检测。
The white rabbits were immuned with the mixtures of the bacteria of Actinobacillus pleuropneumoniae （App） serotype 2 and Freund＇ s incomplete adjuvant. And immunoglobulins（IgGs） purified from the immune sera was to develop double sandwich ELISA for detecting the antibodies of App serotype 2. Checkerboard titrations were performed to determine the optimal concentrations of IgGs coated, antigen and enzyme conjugate. The results of the specific and sensitive tests showed that double sandwich ELISA had the high specificity and sensitivity. It had no cross-reaction with sera from herds infected with FMDV, TGE, PRRS and other A. pleuropneurnoniae serotypes. The coincidence rate of the results of double sandwich ELISA and blocking ELISA reached 99. 54%. The results of this experiment showed that double sandwich ELISA could be used in field of diagnostic work of porcine contagious pleuropneumonia.
出处
《新疆农业大学学报》
CAS
北大核心
2009年第6期46-50,共5页
Journal of Xinjiang Agricultural University
基金
上海出入境检验检疫局项目(HK004-2008)
