ObjectiveTo establish a PCR detection method for simian adenovirus (SAdV) and study the SAdV infection status in monkeys. Methods To design a primer based on SAdV sequences and optimize the PCR experiment, test the method, to detect the virus in monkeys and vaccines, and sequenceing and constructing the phylogeny trees. ResultsA PCR detection method for SAdV was established, which could distinguish SAdV from MAD, ICH, CELO and was sensitive to 47.9 pg/mL viral DNA. The results of this study showed a positive rate of 49.2% and a large genetic diversity of simian adenoviruses in Chinese primate colony. Most of them belong to G subgroup and twig of SAdV-49. ConclusionsThis PCR detection method established in our laboratory has a good accuracy. We found a high prevalence of SAdV in Chinese primate colony. It is necessary to enhance detection and control for SAdV in monkeys and relevant biological products, and avoid potential risk of human infection.