Objective The aim of the present study was to further confirm the viral concentration of SHIV-KB9 virus for infection of Chinese rhesus monkeys，to test the animals' adaptability to the virus，to clarify the repeatability of the animal model，and try to establish a Chinese-origin rhesus macaque model of SHIV virus infection. Methods Four Chinese-origin rhesus macaques were used in this study. All the monkeys were screened by serological test to ensure that they were free from SIV，SRV and STLV infection. Then they were infected by i. v. injection of 10 × diluted viral series separately. Finally to determine if the Chinese-origin rhesus macaques were infected by flow cytometry，blood routine examination，virus isolation and PCR. Results The results of all tests proved that only the virus concentration higher than of 4. 8 × 105 copies/mL can infect the Chinese-origin rhesus macaques. Conclusion This results of this study further clarified the effective concentration of SHIV-KB9 virus for infection of Chinese rhesus monkeys，identified virological and immunological indicator of SHIV-KB9 infection of Chinese rhesus monkeys，and we have successfully established a Chinese rhesus monkeys model of SHIV-KB9 virus infection.