Bananas (Musaceae, Zingiberales) are considered to be one of the most important fruit crops and an important staple food in many developing tropical and subtropical regions (Kumar et al., 2011). In Southern Africa, banana production is under threat from a number of viral diseases, including Banana bunchy top virus (BBTV) (family Nanoviridae, genus Babuvirus). BBTV is the cause of Banana bunchy top disease, the most devastating virus disease of banana. In Sub-Saharan Africa the disease is endemic to 13 countries, including Malawi and Zambia in the Southern African sub-region. Previously, a detection survey for BBTV was done in South Africa in 1996 and this study confirmed the absence of BBTV in the Kiepersol region of Mpumalanga Province (Pietersen et al., 1996). No other surveys were conducted since then in South Africa. The disease is spread by the banana aphid Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), that is present in South Africa, and through infected propagation material. Pentalonia nigronervosa feeds on the phloem of host plants, and potentially picks up the BBTV particles from the phloem and spreads them in plantations as they feed. In June 2015, five 'Williams' plants, showing symptoms resembling BBTV, including streaks on the pseudostem and upright bunchy appearance, were submitted for identification to the Virology Diagnostic Lab at Agricultural Research Council-Plant Protection Research Institute (ARC-PPRI). The plants originated from a farm close to Hibberdene in the KwaZulu Natal South Coast production region (GPS coordinates: S 300 30, 636', E 300 30,653'). A BBTV-specific PCR was done using the protocol published by Thomson and Dietzgen (1995), using primer pairs BBT-1 and BBT-2, amplifying a fragment of the putative replicase gene. The five plants tested positive for BBTV, and the PCR products (~349 bp) of two of the accessions, 15/1408 and 15/1410, were directly sequenced using Sanger sequencing and 321 bp of both sequences were deposited in GenBank (GenBank Accession Nos: KU196167, KU196168). A non-infected banana plant was included in the test as a healthy control. Phylogenetic analyses of the partial replicase genomic region grouped accessions 15/1408 and 15/1410 with the South Pacific group that included accessions from India, Pakistan and other regions in Africa with a sequence identity of 99% (Kumar et al., 2011). A second confirmation test for the presence of BBTV in these plants was conducted using a BBTV-specific Enzyme-linked immunosorbent assay (ELISA) supplied by Agdia (Catalog number: SRA24700) using the manufacturer's instructions. The five plants tested positive for BBTV in the ELISA test, confirming the previous positive results. Forty-four additional samples were included in the ELISA test. These samples were collected randomly from 14 blocks on the same farm (GPS coordinates: S 300 30, 636', E 300 30,653') and comprised of symptomatic and non-symptomatic plants. BBTV was detected with ELISA in 34 of these plants, ten plants tested negative for BBTV. In one of these blocks, C17, all the sampled plants (4), tested negative. In three other blocks (A9, C8 and C15), positive and negative plants were recorded. In the remaining 10 blocks, all collected plants tested positive. Intense management strategies, including removal of infected plants and spraying for aphids, were implemented on the farm to prohibit the spread of the virus to neighbouring plantations. The ELISA test confirmed the earlier detection of BBTV with PCR and sequencing. This is the first report of BBTV on Musa spp. from South Africa. Strict implementation of phytosanitary measures will be necessary to prevent the spread of this virus to other regions in the country. The need to do extensive surveys for the presence of BBTV in other banana production regions in South Africa is highlighted by this finding.