巢式聚合酶链反应快速检测贝氏柯克斯体的研究

目的 建立特异敏感的快速检测贝氏柯克斯体的分子生物学方法。方法 采用巢式聚合酶链反应（ＰＣＲ）方法，进行实验室模拟检测。结果 以贝氏柯克斯体ｃｏｍｌ基因（编码２７０００蛋白）为靶标设计了２对引物；该引物特异性实验表明仅贝氏柯克斯体特异性 段可以扩增出来，其余相关立克次体均扩不出；敏感性测试结果可检测出１．５ｆｇdisplay structure

Nested PCR for rapid detection of Coxiella burnetii

Objective To develop a specific and sensitive molecular biological way for rapid detection of Coxiella burnetii.Method Nested PCR was established and evaluated in the laboratory.Results Two sets of primers targeted to 27 kDa protein gene of C. burnetii were designed. Specific experiment was carried out and the result showed that the primers were highly specific to C. burnetii. Sensitive experiments revealed that 1.5 fg DNA and 0.001 CELD 50 C. burnetii could be detected by the primers. 83.3% C. burnetii could be detected 24 hours after the mice were challenged by 10 000 CELD 50 C. burnetii. The simulated samples of milk, mice spleen and blood, ticks were detected by nested PCR, but the sensitivity decreased when detecting the simulated samples.Conclusion Nested PCR is a specific and sensitive molecular biological way for the rapid detection of C. burnetii.