Methadone is a potent synthetic opioid used in several countries both for substitution therapy in people with opioid dependence and to prevent HIV infection spreading. In Russia methadone trafficking is prohibited. Despite the current ban, cases of hospitalization with methadone poisoning are regularly recorded in an intensive care units and toxicological departments of emergency hospitals (TD EH). Frequently polydrug consumption detected in a laboratory study. People who use psychoactive substances (PS) are vulnerable to HIV-infection as well as hard-to-reach group for HIV testing. Often HIV-infection and drug abuse pathology are revealed only when drug users hospitalized to an emergency hospital after overdose with one or several PS. During the period 2015–2018 to the toxicological department of Sklifosovsky Research Institute for Emergency Medicine hospitalized 732 patients with acute methadone poisoning. In 76,8–83,9% of cases, methadone was detected in combination with other PS, predominantly with opiates/opioids, psychostimulants, psychodysleptics, as well as pharmaceutical medicaments. In addition to methadone, we recorded the presence of ethanol in biological samples (blood and urine) in a concentration from 0,3 to 3,6‰ in 19,2–33,3% of cases. The prevalence of HIV infection in patients with acute methadone poisoning ranged from 15,7 to 31,8%, increasing by 1,5 times during the analyzed period. The reasons for the increase of prevalence HIV infection in hospitalized patients could be linked to a change in the structure of the taken PS and virus transmission pathways. Patients of TD EH are a high-risk group for the spread of HIV and can be considered as a focus group reflecting general trends in the consumption of drugs in Moscow.