The Tibet Autonomous Region (referred to as'Tibet') is located in southwestern China, bordering the Yunnan Province, Sichuan Province, Xinjiang Uygur Autonomous Region, and other provinces with high HIV/AIDS prevalence, as well as India, Nepal, and other high-risk border countries. In 1994, the first patient with HIV-1 infection was detected at Zhangmu port, Namu County, Shigatse, Tibet. In 2005, the first AIDS cases were reported among local Tibetans. A total of 253 cases of HIV/AIDS were reported between 2004 and 2013. The number of HIV/AIDS cases in Tibet is increasing each year, and this epidemic encompasses seven cities throughout the region. Even though the overall epidemic has a low countrywide prevalence, the prevention and control of HIV-1 in Tibet cannot be ignored. Previous studies on patients with HIV-1 in Tibet have mainly focused on epidemiological studies, and to the best of our knowledge, no relevant study has reported the genotype of HIV-1. Therefore, this study aimed to analyze the genotype and source of HIV-1 infection strains in Tibet.