Despite the implementation of health education and free condom distribution for decades, the HIV/AIDS epidemic among older adults in China shows no sign of declining. This study aims to identify HIV transmission patterns and pathways in a rural county area and provide insight for developing effective HIV prevention strategies among older adults. Epidemiological field surveys combined with phylogenetic analysis were used to identify potential HIV transmission linkage in one rural county with a rapidly increasing HIV epidemic among older adults. A total of 160 HIV-positive individuals and their HIV-positive sexual partners diagnosed between 2015 and 2018 were recruited. Among them, 69.4% ( n = 111) were diagnosed at age 50 or older, 68.1% ( n = 109) were men, and 55.0% ( n = 88) were married. Ninety-six participants self-reported as having been infected with HIV through commercial heterosexual behavior and 24 were infected from their HIV-positive spouse. CRF08_BC (67.0%, n = 73) and CRF07_AE (19.3%, n = 21) were the prevalent HIV stains and formed 15 clusters at distance cutoff of 0.0025. Participants diagnosed with HIV at age ≥50, those who were divorced or bereft their spouse, or infected with HIV locally were more prevalent within the 15 molecular clusters. We identified specific venues as the primary hotspot and found that commercial heterosexual contact between older men and commercial sex workers contributed to the local HIV epidemic. In rural areas with an increasing HIV epidemic, detecting HIV transmission clusters through epidemiological and phylogenetic analysis as well as designing localized HIV prevention strategies should be prioritized.

HIV drug resistance compromises the ability of anti-retroviral therapy (ART) to suppress viral replication, resulting in treatment failure. This study investigates the prevalence of pre-treatment drug resistance (PDR) in newly diagnosed individuals in a prosperous city (Wenzhou) in Southeastern China. A cross-sectional investigation was carried out among 473 newly diagnosed ART-naive HIV-1-infected individuals between January and December 2022. The protease–reverse transcriptase (PR-RT) region and integrase (IN) region of HIV-1 were amplified by two separately nested PCRs, followed by sequencing. Drug resistance mutations (DRMs) and drug resistance to nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors (PIs) and integrase strand transfer inhibitors (INSTIs) were analyzed. The PDR prevalence was 6.5% [95% CI: 4.4–9.1] for any anti-retroviral drug, 0.9% [95% CI: 0.3–2.3] for NRTIs, 4.1% [95% CI: 2.5–6.5] for NNRTIs, 1.8% [95% CI: 0.8–3.6] for PIs and 0.5% [95% CI: 0.1–1.8] for INSTIs. According to the subtyping results of the PR-RT region, 11 different subtypes and 31 unique recombinant forms (URFs) were found. CRF07_BC was the dominant subtype (53.7%, 233/434), followed by CRF01_AE (25.3%, 110/434). V179D (1.6%) and K103N (1.4%) were the most predominant types of NNRTI DRMs. Q58E (1.2%) and M184V (0.7%) were the most frequent PI DRMs and NRTI DRMs, respectively. The INSTI-related DRMs Y143S (causes high-level resistance to RAL) and G163K (causes low-level resistance to EVG and RAL) were found in one patient each. Given the relatively high PDR prevalence of NNRTI (4.1%), non-NNRTI-based ART may be preferred in the future. It is recommended to include genotypic resistance testing before starting ART in regions where feasible.