动物医学进展2024，Vol.45Issue(7)：6-10,5.
羊贝氏柯克斯体TaqMan荧光定量PCR检测方法的建立及应用
Establishment and Application of Taq Man Real-time PCR Method for Detecting Coxiella burnetii
摘要
Abstract
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.关键词
贝氏柯克斯/TaqMan荧光定量PCR/检测方法Key words
Coxiella burnetii/Taq Man real-time PCR/detection method分类
农业科技引用本文复制引用
张锐铮,于皓同,王凯茸,张琪,张淑霞,许信刚..羊贝氏柯克斯体TaqMan荧光定量PCR检测方法的建立及应用[J].动物医学进展,2024,45(7):6-10,5.基金项目
国家重点研发计划项目(2021YFD1600704) （2021YFD1600704）
陕西省农业专项资金项目项目畜牧专项(XNDY2210) （XNDY2210）
