液相蛋白芯片同分型检测猪传染性胸膜肺炎诊断方法的研究.pdfVIP

液相蛋白芯片同步分型检测猪传染性胸膜肺炎诊断方法的研究 摘 要 猪传染性胸膜肺炎（Porcine contagious pleuropneumoniae ）是由胸膜肺炎放线杆菌 （APP ）引起的一种猪的呼吸道传染病。对于 APP 的检测，国内外已报道了许多种方法， 像病原分离鉴定、血清学方法和分子生物学技术等，这些方法可以很好的检测出 APP ， 但存在着一定的缺点，有的只能确定病原不能具体区分血清型，如病原分离培养；有的 实验操作很繁琐，需要专业的技术人员，如补体结合试验，等等。本研究建立的液相蛋 白芯片方法，可以同时检测猪血清中六种血清型的 APP 抗体。检测猪病毒、细菌性传 染病如猪 O 型口蹄疫、猪传染性胃肠炎、蓝耳病等阳性血清，结果均为阴性。对 218 份临床猪血清检测，比较其与 CF 试验在检测 APP 不同血清型抗体的结果，液相蛋白芯 片检测结果与 CF 试验检测猪血清样本结果相符率为 98.16%，在统计学上对猪血清样本 判断无明显的差异(P0.05) 。 本研究将液相芯片技术应用于猪胸膜肺炎放线杆菌的分型诊断中，在后续研究中， 该技术可一次同时检测 15 个血清型的 APP ，大大减少了工作量，同时具有快速、准确、 敏感、特异等优点，对我国猪传染性胸膜肺炎的进出口检疫有很大的实际应用价值。 关键词：猪传染性胸膜肺炎；双夹心 ELISA ；液相蛋白芯片；同步检测 I Research on Simultaneous Detection of Antibodies to Different Serotypes of Actinobacillus Pleuropneumoniae by Liquid Protein Chips Abstract Porcine contagious pleuropneumoniae is an porcine respiratory passage infectious diseases caused by Actinobacillus pleuropneumoniae (APP).Many methods detecting APP had been reported in domestic and abroad ，such as pathogeny dysjunction；serological method； molecular biology technique.those methods can be used detecting APP widely ，but it existed unvarying disadvantage. The ltaneous detect antibodies to six kinds of serotypes of A. pleuropneumoniae can be detected by the liquid protein chips developed in this text. While the results of other antibodies such as FMD ，TGE，PRRS were observed negatively. It had no cross-reaction with sera from herds infected with other A. pleuropneumoniae serotypes. 218 share pig clinical sera were dected to compare the antibody of different serological type by CF test. the analyzed results from liquid protein chips were identical with those from complement fixation test(CF) basic