Abstract: Objective To investigate the current status of HIV-related health service uptake among cross-border couples (Chinese husbands and Burmese wives) in Dehong Dai and Jingpo Autonomous Prefecture, and to provide evidence for optimizing HIV prevention and control strategies in border areas. Methods A cross-sectional survey was conducted from September 2022 to September 2023 among 14 662 cross-border couples (Chinese husbands and Burmese wives) in Dehong Prefecture. Data on sociodemographic characteristics, HIV/AIDS knowledge awareness, and utilization of HIV-related health services within the past year were collected. Multivariable logistic regression was used to analyze factors associated with HIV testing uptake among husbands and wives. Results A total of 29 324 individuals (14 662 couples) were included. The overall awareness rate of HIV/AIDS was 82.09%, with the rate among husbands significantly higher than that among wives (85.17% vs 79.01%, P<0.001). Overall, 94.98% of participants had received at least one HIV-related health service, and the HIV testing rate in the past year was 65.52%. Multivariable analysis showed that, compared with participants married for 0-3 years, those married for 4-10 years and ≥ 11 years were more likely to have received HIV testing. The corresponding ORs were 1.344 (95% CI: 1.187-1.521) and 1.381 (95% CI: 1.211-1.574) among husbands, and 1.308 (95% CI: 1.155-1.481) and 1.344 (95% CI: 1.178-1.533) among wives, respectively. Compared with those holding a marriage certificate, those possessing a border marriage registration certificate were positively associated with HIV testing uptake with ORs of 1.710 (95% CI: 1.575-1.857) among husbands and 1.700 (95% CI: 1.565-1.847) among wives. Compared with participants who were illiterate, those with primary school education showed lower testing rates, with ORs of 0.738 (95% CI: 0.650-0.837) among husbands and 0.827 (95% CI: 0.766-0.891) among wives. Additionally, a history of sexually transmitted diseases (OR=3.66, 95%CI: 1.24-15.59) and participation in the New Rural Cooperative Medical Scheme (OR=1.51, 95%CI: 1.18-1.94) were independently associated with higher testing rates among husbands. Conclusion The overall utilization of HIV-related health services among cross-border couples in Dehong Prefecture is generally satisfactory, but significant gender disparities exist, with foreign wives showing lower rates of knowledge awareness and health service utilization compared with their Chinese husbands. Couples with shorter marriage duration, with primary school education, and those not included in the border marriage registration system are vulnerable groups for HIV testing services. Future interventions should prioritize cross-border marriage families at the early stage of entry and in the newly married period. Based on the border marriage registration management system and primary healthcare networks, health education and HIV testing services targeting foreign spouses should be strengthened to improve service accessibility and equity.