Population movement dynamics are a critical part of understanding communicable disease transmission patterns and determining where, when, and with whom to deliver appropriate prevention interventions. This study aimed to identify the origin of the Afghan population and their patterns of movement within Karachi, to assess the polio vaccination status of children under the age of five, and to investigate the travel history and guest arrival patterns of individuals from Afghanistan and other regions known to be affected by wild poliovirus type 1 (WPV1) within the past six months. A cross-sectional survey was conducted in selected 12 union councils of Karachi, Pakistan. The data were collected through interviews with Afghan household members and from the frontline workers (FLWs) responsible for the polio vaccination of the children of the same households. Cohen's kappa was used to check the agreement between information provided by the household participant and FLWs. A total of 409 Afghan household members were interviewed. Travel of any household member outside the city within the last six months was reported by 105 (25.7%) participants, 140 (34.2%) hosted guests within the last six months, and 92 (22.5%) participants reported that guest children were vaccinated in their households. A total of 230 (56.2%) participants observed polio teams at relatives' households within Karachi, and 127 (31.1%) observed polio teams at relatives' households outside Karachi in different districts of Pakistan and Afghanistan. Fair to moderate agreement was observed between information provided by the household members and FLWs on the variable's duration of living at current residence (Kappa = 0.370), travel history (Kappa = 0.429), guest arrival (Kappa = 0.395), and household children vaccinated for OPV (Kappa = 0.419). Substantial population mobility was observed between Afghanistan and Pakistan as well as significant movement of the Afghan population within Karachi in the last six months. These findings warrant attention and targeted implementation of interventions to enhance and sustain both routine and supplementary immunization activities within this demographic group.