The continued endemic presence of measles virus (MV), and the large number of isolates which are made in South Africa each year, demanded the use of a rapid and reliable pre-screening technique to select isolates for molecular epidemiological studies by sequence analysis. The heteroduplex mobility assay (HMA) was used to genetically characterize 47 MV isolates collected from three different provinces in South Africa, made between 1986 and 1995. The carboxyl-terminal 590 nt of the nucleocapsid (N) gene--the most variable region of the genome--was amplified by polymerase chain reaction (PCR) and subsequently subjected to HMA analysis for initial genotyping. The results showed three different patterns of heteroduplex formation by gel electrophoresis, representing two distinct wild-type lineages and one group of vaccine-like viruses. Comparison of HMA results with phylogenetic analysis of sequence data for several of the South African MV strains showed a complete correlation of results. The HMA proved to be a useful tool for screening MV isolates for use in molecular epidemiological studies.