Camel pox is a viral contagious notifiable zoonotic disease causing variable rates of mortality, morbidity and cutaneous lesions in camels and humans. It caused by camelpox virus (CMLV), an orthopoxvirus and can be fatal in young camels. Confirmative, rapid and sensitive diagnostic techniques are required for accurate diagnosis of the disease, so in the present study, a duplex real-time PCR assay was developed for rapid detection of camelpox virus (CMLV) based on targeting both C18L and DNA pol genes. The detection limit was 0.4 pg and 4 pg of viral DNA for C18L and DNA pol genes, respectively. The duplex assay detected CMLV DNA in different types of spiked samplesand providesa rapid diagnostic tool for camel pox especiallyin borders where testing of large number of animals during importation and exportation is applied. Further evaluation of the assay on other clinical samples or samples from experimentally infected animal may be required.