Molecular detection of human immunodeficiency virus RNA in Maryland wastewater

Human Immunodeficiency Virus (HIV) is still one of the biggest global public health issues, and the disease burden is disproportionately high in areas with lower socioeconomic status. Research has demonstrated that even in patients on antiretroviral medication, HIV RNA can be excreted in urine and feces. However, the use of wastewater-based epidemiology (WBE) for HIV surveillance has not yet become widespread. The goals of this study were to detect HIV RNA in raw wastewater and analyze its spatial patterns in relation to socioeconomic levels in Baltimore City and the surrounding areas. 104 raw wastewater samples were collected from six wastewater treatment facilities across the state of Maryland from February 2024 through April 2025. Samples were concentrated using polyethylene glycol (PEG 8000), and qPCR was used to measure the levels of HIV in wastewater. Pepper mild mottle virus (PMMoV) was used as an internal process control. HIV RNA concentrations, spatial distribution, temporal volatility, and correlations with socioeconomic variables were examined and contrasted with results from earlier research. All samples had PMMoV, indicating little PCR inhibition and excellent extraction quality. HIV RNA was found in 12.5 % of samples (13 out of 104), and the majority of positive results came from 2 treatment facilities that served urban areas that were highly inhabited and socioeconomically disadvantaged. This is the first study to report the detection of HIV RNA in raw wastewater in Maryland. Cost-effective, scalable, and non-invasive, the suggested method provides a potent tool for targeted surveillance and early identification in regions most in need of public health interventions, especially considering the increased prevalence of this virus in underprivileged communities.