羊贝氏柯克斯体TaqMan荧光定量PCR检测方法的建立及应用OA北大核心CSTPCD

To establish the TaqMan real-time PCR detection method for Coxiella burnetii,we referred to the C.burnetii included in GenBank.We used the conserved region of the RSA439 strain com1 gene se-quence(CP040059.1)to design specific primers and probes,construct recombinant plasmids,and optimize reaction conditions in order to establish a Taq Man real-time PCR method for Coxiella burnetii.The results showed that the standard curve established by this method had a good linear relationship,and the sensitivi-ty results showed that the minimum detection limit of fluorescence quantitative PCR was 2.45 × 102 copies/μL,which was 100 times more sensitive than 2.45×104 copies/μL.This method showed no cross-re-action to Brucella,Chlamydia,and Mycoplasma.The results of intra-and inter-batch tests were stable,with the coefficient of variation less than 3％,indicating that the method had good specificity and was re-producible.The method was used to test 126 collected samples and showed a positive rate of 9.52％,com-pared to 7.14％by conventional PCR,with a compliance rate of 90.47％.This method provides technical support for the rapid diagnosis and prevention of clinical diseases.