Development of new multiplex nested PCR for rapid detection of Coxiella burnetii

Q fever is a significant worldwide zoonosis caused by Coxiella burnetii, an obligate intracellular, gram-negative and pleomorphic rod. This bacterium caused one type of foodborne diseases and the prevalence of food poisons has increased recently, therefore an accurate diagnosis of this pathogen is necessary. In this study, three sets of specific primers were designed with the goal of multiplex nested PCR detection of C.burnetii. Com1 and IS1111 repetitive elements were chosen for multiplex nested PCR assay. After that, PCR products cloned into the TA vector in order to design a PCR positive control construct. PCR reaction was performed and three specific bands at 139, 300 and 598 bp were observed in 2% agarose gel electrophoresis stained with KBC power load. The sensitivity of this method was determined about 2.1x10(-2) pg for com1 and 12 fg for IS1111. Development of this new PCR reaction is useful for a simple, rapid and correct diagnosis of C.burnetii in food and dairy product. To conclude, this new multiplex nested PCR assay could be used for identification of this important pathogen in research laboratories without culture-based equipment.