Rapid Detection of HIV-1 Subtypes in Ghana by Heteroduplex Mobility Assay

Abstract

Background: In Ghana, the HIV-1 profile has been quite dynamic. Previous reports identified HIV-1 subtypes A, D and G present and recently the CRF02_AG has been described as the predominant molecular form of HIV-1 in Kumasi, Ghana. This underscores the need for constant molecular characterization of HIV-1 species in the country. Objective: To provide current updates on the nature of HIV subtypes in Ghana, there is need for a user-friendly tool for routine monitoring of subtypes in the absence of cost-intensive and skill-demanding sequencing techniques. This work demonstrates the use of heteroduplex mobility assay (HMA) for rapid subtype detection of HIV-1 isolated from Ghanaian patients. Method: Viruses from 15 antibody-positive HIV-1 patients were isolated directly by co-culturing peripheral blood mononuclear cells (PBMC) with phytohemagglutinin (PHA)-stimulated donor PBMCs from an HIV seronegative individual and through HeLa cells positive for CD4/CXCR4/CCR5 (MAGIC-5A cells). That was followed by proviral DNA extraction. Heteroduplex Mobility Assay (HMA) technique was then performed on the HIV gag gene. Results: Using the HMA technique, newly isolated HIV-1 strains were subtyped as follows: There were seven subtype A (47%), two subtype G (13%) and six (40%) A/G recombinants. Conclusion: The HIV-1 CRF02_AG in Ghana has spread much more rapidly than the previously predominant subtype A over the years. Constant molecular characterization of HIV strains is necessary to enable clear elucidation of the prevailing HIV species in Ghana. This study presents the HMA as a useful tool for monitoring subtype emergence and distribution in the country.