| Summary: | Assessment of the long-term population-level effects of HIV interventions is an ongoing public health challenge. Following the implementation of a Transmission Reduction Intervention Project (TRIP) in Odessa, Ukraine, in 2013–2016, we obtained HIV <i>pol </i>gene sequences and used phylogenetics to identify HIV transmission clusters. We further applied the birth-death skyline model to the sequences from Odessa (<i>n </i>=<i> </i>275) and Kyiv (<i>n </i>=<i> </i>92) in order to estimate changes in the epidemic’s effective reproductive number (<i>R</i><sub>e</sub>) and rate of becoming uninfectious (<i>δ</i><i>)</i>. We identified 12 transmission clusters in Odessa; phylogenetic clustering was correlated with younger age and higher average viral load at the time of sampling. Estimated <i>R</i><sub>e</sub> were similar in Odessa and Kyiv before the initiation of TRIP; <i>R</i><sub>e</sub> started to decline in 2013 and is now below <i>R</i><sub>e </sub>=<i> </i>1 in Odessa (<i>R</i><sub>e</sub> = 0.4, 95%HPD 0.06–0.75), but not in Kyiv (<i>R</i><sub>e</sub> = 2.3, 95%HPD 0.2–5.4). Similarly, estimates of <i>δ</i> increased in Odessa after the initiation of TRIP. Given that both cities shared the same HIV prevention programs in 2013–2019, apart from TRIP, the observed changes in transmission parameters are likely attributable to the TRIP intervention. We propose that molecular epidemiology analysis can be used as a post-intervention effectiveness assessment tool.
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