Dot-PPA-ELISA联合检测猪PRV.Brucella和Chlamydia抗体方法的建立和应用研究

首次将Dot-PPA -ELISA用于联合检测猪衣原体、伪狂犬病毒和布氏杆菌抗体 ,并进行了较大规模的田间猪血清检测。实验对制作诊断膜片的工艺流程和关键材料做了细致研究 ,并进行同步监测、分析。确定了本方法的最佳反应条件和阳性标准。联合诊断膜片 (简称膜片 )具有良好特异性、灵敏性和稳定性。诊断膜片在 4℃保存 8个月、室温 (15～ 2 5℃ )保存 2个月、37℃保存 1个月灵敏性 ,特异性不变。 1∶6 4、1∶12 8、1∶6 4为联合检测猪伪狂犬病、衣原体病和布氏杆菌病抗体的阳性标准。结果表明 ,Dot -PPA -ELISA联合检测法操作方便、快速 ,结果客观 ,易于判读 ,诊断膜片便于寄送、保存 ,性能稳定、可靠 ,并能同时检测多种疫病等优点 ,适宜应用于猪衣原体病、伪狂犬病和布氏杆菌病抗体的联合检测

Studies on A Dot-PPA-ELISA for the Co-detection of PRV,Brucella and Chlamydia Antibodies and it's Appliance

A Dot-PPA-ELISA was developed for the co detection of swine Brucella, PRV and Chlamydia antibodies for the first time and the capability of it's relatively large scale appliance was determined. In this study was careful study carried through in technecs flow of diaphragm making and the key matiarial. The best reacting condition and other important reagent were determined. The co detection diaphragm does not react with the antibodies of swine fever, swine foot mouth disease and swine bow-shaped body disease. The minium amount of Prv, Brucella and Chlamydia serum IgG giving a positive reaction is 5 326×10 -9 g, 7 221×10 -9 g and 1 21×10 -9 g. 12 serum samples are detected for 3 times, making T test, the result is P>0 05. The diaphragm stored at room tenperature for 2 monmths, at 4℃ at least for 8 months, at 37℃ for 1 months, the sensitivity and specificity did not change. The positive standards of the diaphragm of Prv, Brucella and Chlamydia antibodies were 1∶128, 1∶64 and 1∶64. All the results show the co detection diaphragm is specific, stable, rapid detection. It can be used in co detecting the Prv, Brucella and Chlamydia antibodies.