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目的 探讨用防污染的 PCR- EL ISA(U DPE)技术检测人为污染的土壤和感染的动物标本的可行性。方法 根据鼠疫耶尔森氏菌 Cafl和 Pla基因分别设计了 3条引物 ,组合成 3对引物 ,建立 U DPE检测技术。结果 利用该实验体系可以检出每克土壤中 10 0 0个鼠疫耶尔森氏菌的污染。比检测纯细菌的敏感性低 2 0 0倍 ,说明土壤中有抑制 PCR反应的因子。对 2 0只实验组动物 40份标本 (肝脏和脾脏 )的培养、U DPE和 PCR-电泳检测表明 ,3种方法的检出吻合率为 10 0 % ,均能从 40份标本中检出靶细菌 ,而用 3种方法检测对照组动物的 2 0份肝脏和脾脏均为阴性。结论 利用 UDPE技术可以被用于动物脏器中鼠疫耶尔森氏菌的检测
Objective To explore the feasibility of detecting contaminated soils and infected animal specimens with PCR-EL ISA (U DPE). Methods According to the genes of Cafl and Pla of Yersinia pestis, three primers were designed respectively and combined into three pairs of primers to establish U DPE detection technique. Results The system was used to detect 10 0 Yersinia pestis contamination per gram of soil. Lower than the detection of pure bacteria sensitivity 200 times, indicating that inhibition of soil PCR reaction factor. The culture of 40 specimens (liver and spleen) of 20 experimental animals was detected by U DPE and PCR-electrophoresis. The coincidence rate of the three methods was 100%, and the detection target was 40 specimens Bacteria, while 20 animals in the control group were negative for both liver and spleen in 3 ways. Conclusion The UDPE technique can be used to detect Yersinia pestis in animal organs