Entry of human immunodeficiency virus-1 (HIV-1) into host cells involves an interaction between host cell receptors and the virus envelope glycoproteins, specifically, glycoprotein 120 (gp120). The variable 3 (V3) loop region of HIV-1 gp120 envelope protein plays an essential role in HIV-1 coreceptor binding and is thus a determinant for host cell tropism. Viral tropism is therefore an important consideration during coreceptor-dependent therapy development, necessitating the current study. Venous blood samples were collected and plasma obtained from 110 patients attending HIV Clinics in 3 hospitals in Kiambu County. RNA extraction was done, followed by reverse transcriptase polymerase chain reaction (RT-PCR) and Sanger sequencing. Six sequences (N=6) were generated for downstream analysis, and all were determined to be subtype A1 using phy-logenetic analysis. To validate the study findings due to the few sequences, an additional 62 HIV-1 subtype A1 sequences from the HIV Los Alamos National Library (LANL) database were extracted and analysed with the study sequences for tropism and genetic diversity validation of the V3 loop region. Five of the six study sequences were identified as CXCR4-tropic strains. The V3 region comprising 35 amino acids harboured a conserved N-linked glycosylation site at amino acid position 301 in all the sequences. The V3 crown region was conserved in all the sequences except for the CCR5 tropic strain that harboured mutations. Although HIV-1 subtype A1 showed high CCR5 usage on validation, CXCR4 usage was common among the study sequences. CCR5-using strains were likely to harbour V3 crown mutations in this subtype. Further research to understand the genetic diversity of subtype A1 V3 region is advocated.