HIV epidemic in Pakistan is characterized by high prevalence in IDUs and Male and Transgender sex workers but very low prevalence in Female sex workers. Most of females have acquired HIV from their husband.Out of 556 Afghan Refugees 5.9% were HIV, 36.8% HCV and 9.17% HBV positive. Of 396 MTSW 11.6% HIV, 20.7% HCV and 7.6% were HBV positive. Among 28 Afghan HIVs 17 were subtype A1, 05 aligned with CRF 01_AE (CRF 01_AU) and 06 with 02_AG. Out of 33 MTSW HIVs 21 were subtype A1, 02 were G and 10 were CRF 01_AE. All the subtype A1 strains and CRF 01_AE show very close homology (Founder effect). The present study has for the first time reported subtypes G, Circulating Recombinant Form (CRF) 01_AE and CRF 02_AG Other HIVs were subtype A 62%, CRF 01_AE 25%, CRF 02_AG 10% and subtype G 3%. Contrary to the earlier opinion that HIV in Pakistan was a monophyletic epidemic brought by returning expatriate workers from gulf states the detection of several subtypes have proved that the HIV epidemic in Pakistan is polyphyletic event and HIVs from different parts of the world have been involved in rapid spread of HIV, wide clinical spectrum and highly variable ART response.