从动物和土壤中直接检测鼠疫耶尔森氏菌

Direct detection of Yersinia pestis from animal and soil

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

Objective An anti contamination PCR ELISA technique (UDPE) was employed for direct detection of Yersinia pestis from artificially contaminated soil and infected mice.Methods Primers used in this system were designed according to Cafl and Pla genes of Y.pestis.Three combinations of these primers were used to establish UDPE technique for detecting Y.pestis from the mock samples.Results One thousand of bacteria in one gram soil could be directly detected by this system,which is 200 times lower than those in bacterial suspension.This indicated that there were PCR inhibitors in soil.Forty specimens(liver and spleen) from laboratory infected mice were examined by conventional culture method,UDPE and PCR gel electrophoresis simultaneously,and the result showed that Yersinia pestis in the sample could be detected by all three methods.Conclusions Our results demonstrated the feasibility of UDPE for direct detecting Y.pestis in animal but the soil processing method needed to be improved. [