Objective To investigate the status of Helicobacter pylori and "Helicobacter macacae" infection in rhesus and cynomolgus monkeys in China. Methods With the use of 16S rRNA specific primers for Helicobacter spp and Helicobacter pylori (HP) from published literatures, and new 16S rRNA specific primers designed for "Helicobacter macacae" (HM), we investigated the infection status of these two Helicobacter spps in both of 45 rhesus and 90 cynomolgus monkeys by qPCR or conventional PCR on stool samples. Results All three primer sets for 16S rRNA exhibited excellently sensitivity and specificity. Both the infection rates of HP and HM were 100% among 45 young adult rhesus monkeys. The infection rate of HP and HM in 90 young adult cynomolgus monkeys were 100% and 97.8%, respectively. Conclusions Helicobacter pylori and "Helicobacter macacae" are present in almost every artificially bred adult rhesus and cynomolgus individuals which may adversely affect the health of laboratory monkeys and the accuracy of related animal experiments.