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<i>Rickettsia</i> species are an important cause of emerging infectious diseases in people and animals, and rickettsiosis is one of the oldest known vector-borne diseases. Laboratory diagnosis of <i>Rickettsia</i> is complex and time-consuming. This study was aimed at developing two quantitative real-time PCRs targeting <i>ompB</i> and <i>ompA</i> genes for the detection, respectively, of <i>Rickettsia</i> spp. and <i>R. conorii</i> DNA. Primers were designed following an analysis of <i>Rickettsia</i> gene sequences. The assays were optimized using SYBR Green and TaqMan methods and tested for sensitivity and specificity. This study allowed the development of powerful diagnostic methods, able to detect and quantify <i>Rickettsia</i> spp. DNA and differentiate <i>R. conorii</i> species.