Berger Sanne Schou, Lauritsen Klara Tølbøll, Boas Ulrik, Lind Peter, Andresen Lars Ole
Section for Diagnostic and Scientific Advice, National Veterinary Institute, Technical University of Denmark, Kgs. Lyngby, Denmark.
J Vet Diagn Invest. 2017 Nov;29(6):797-804. doi: 10.1177/1040638717719481. Epub 2017 Jul 7.
We developed and made a preliminary validation of a bead-based multiplexed immunoassay for simultaneous detection of porcine serum antibodies to Actinobacillus pleuropneumoniae serovars 1, 2, 6, 7, and 12. Magnetic fluorescent beads were coupled with A. pleuropneumoniae antigens and tested with a panel of serum samples from experimentally infected pigs and with serum samples from uninfected and naturally infected pigs. The multiplex assay was compared to in-house ELISAs and complement fixation (CF) tests, which have been used for decades as tools for herd classification in the Danish Specific Pathogen Free system. Assay specificities and sensitivities as well as the corresponding cutoff values were determined using receiver operating characteristic (ROC) curve analysis, and the A. pleuropneumoniae multiplex assay showed good correlation with the in-house ELISAs and CF tests with areas under ROC curves ≥ 0.988. Benefits of multiplexed assays compared to ELISAs and CF tests include reduced serum sample volumes needed for analysis, less labor, and shorter assay time.
我们开发并初步验证了一种基于磁珠的多重免疫分析方法，用于同时检测猪血清中针对胸膜肺炎放线杆菌1、2、6、7和12型血清型的抗体。将磁荧光珠与胸膜肺炎放线杆菌抗原偶联，并使用一组来自实验感染猪的血清样本以及未感染和自然感染猪的血清样本进行检测。将该多重分析方法与内部酶联免疫吸附测定（ELISA）和补体结合（CF）试验进行比较，在丹麦无特定病原体系统中，几十年来一直将ELISA和CF试验用作畜群分类工具。使用受试者工作特征（ROC）曲线分析确定分析方法的特异性和敏感性以及相应临界值，胸膜肺炎放线杆菌多重分析方法与内部ELISA和CF试验显示出良好相关性，ROC曲线下面积≥0.988。与ELISA和CF试验相比，多重分析方法的优势包括分析所需血清样本量减少、劳动力减少以及分析时间缩短。
