Circulating recombinant form (CRF) 16_A2D is a rare HIV strain among the most recently identified subsubtype A2. Samples taken from an HIV-seropositive married couple who attended a voluntary counseling and testing unit in Dhaka, Bangladesh showed two rare CRF16_A2D isolates. Further genetic analyses targeting three HIV genes (gag, pol, and env) showed that the two isolates from the couple were closely related making it likely that one of them acquired the virus and transferred it to the other. The husband admitted that he had contact with sex workers and both wife and husband had never traveled outside the country. This indicates that the husband might have acquired the virus from sex workers and that this rare strain is already circulating in the country. Detecting unusual isolates is important to determine the true diversity of HIV strains in the country. Monitoring of such strains will help in identifying transmission patterns and possible interventions to prevent the spread of HIV infection.