HIV-1 subtype C is the major subtype in India as evidenced from the analysis of specific regions within envelope and gag gene. The matrix protein (p17) of HIV-1 which is involved in several functions like the viral RNA transport, nuclear localization, assembly of pre-integration complex into host nucleus has been used to study the strain diversity among female sexworkers in Calcutta. The gene encoding for the HIV-1 matrix protein, p17 was amplified by nested polymerase chain reaction (PCR) from blood samples of HIV-1 seropositive female sex workers (FSW) in Calcutta, India. Genes of twenty-two samples were sequenced and the phylogenetic analysis with different global strains showed that the majority (seventeen) was clustered with Indian type C. A few samples were found to be close to other C subtypes isolated from South Africa, China and Myanmar. The comparison of Calcutta samples with the samples from other regions of India along with other non-asian subtype C sequences clearly revealed a different cluster of Indian sequences. The two samples, cal 242 and cal 709 was found to be the most divergent type and showed close relatedness with African C subtypes.