用液相芯片方法检测猪传染性胸膜肺炎S1血清(亚)型抗体的研究.pdfVIP

上海市畜牧兽医学会 2011 年学术年会论文集 用液相芯片方法检测猪传染性胸膜肺炎S1 血清（亚）型抗体的研究 李树清，王 艳，张 强，王巧全，陈志飞 （上海出入境检验检疫局，上海 200135 ） 摘 要：目的 建立用液相芯片技术快速检测猪AppS1 血清型的新方法，并对该方法进行评价。 方法 制 备抗体交联微球及生物素标记抗体，用双抗体夹心原理，建立液相芯片技术快速检测猪AppS1 血清型的方 法，检测方法的特异性、敏感性，确定阴、阳性判定阈值。用建立的方法测定临床样品，并与商品化ELISA 试剂盒检测结果进行比较。 结果 该方法能够特异性地检测出S1 样品，检测信号具有较高的荧光强度，对 其他血清型（S2-7 ）和相关猪病阳性血清检测结果为阴性。该方法检测灵敏度高于现有商品化App S1 ELISA 检测试剂盒，同时该方法具有较好的重复性，其变异系数为 14.7%。 结论 利用液相芯片技术研制的猪 App 抗体检测方法能够用于AppS1 亚型的快速检测及鉴定，并为进一步联合检测S1-15 型奠定了基础。 关键词：液相芯片技术；App ；S1 亚型 Study of Detecting APP S1 Serotype By Using Liquichip Li Shu-qing, Wang Yan, Zhang Qiang, Wang Qiao-quan, Chen Zhi-fei (Shanghai Entry-Exit Inspection and Quarantine, Shanghai, 200135) Abstract: To establish a new assay for the rapid determination of App S1 basing on the multiplexed Luminex assay and evaluate its assay performance. Method: Antibody-conjugated microspheres and biotinylated detection antibodies were prepared. A sandwich immunoassay was established to measure the samples. Results: This assay could be used for the rapid and specific determination of App S1. High fluorescence intensity was detected in S1 sample but not in other samples of S2-7 and control samples. And the results of sensitive detection also showed that this assay was more sensitive than commercial ELISA Kit. This assay has a reasonable repeatability, the variable factor is 14.7%. Conclusion: The multiplexed Luminex based determination method could be used for high-specific, sensitive and rapid determination of App S1, and this assay also provided a foundation for further research on rapidly high throughout detection for S1-15 serotypes of App. Key words: Liquichip detection technique; App; S1 serotype 猪传染性胸膜肺炎(Infectious pleuropneumonia of swine, IPPS )是由胸膜肺炎放线杆菌 （Actinobacillus pleuropneumonia, Ap