The Girlhood of Mary Virgin is an 1849 oil on canvas painting by the Pre-Raphaelite Brotherhood painter Dante Gabriel Rossetti, measuring 83.2 by 65.4 cm and now in the collection of Tate Britain, to which it was bequeathed in 1937 by Agnes Jekyll. It was his first completed oil painting and is signed "Dante Gabriele Rossetti P.R.B. 1849". He first exhibited it at the 'Free Exhibition' at the Hyde Park Corner Gallery.

The Pre-Raphaelite Brotherhood was a group of English painters, poets, and art critics, founded in 1848 by William Holman Hunt, John Everett Millais, Dante Gabriel Rossetti, William Michael Rossetti, James Collinson, Frederic George Stephens and Thomas Woolner who formed a seven-member "Brotherhood" partly modelled on the Nazarene movement. The Brotherhood was only ever a loose association and their principles were shared by other artists of the time, including Ford Madox Brown, Arthur Hughes and Marie Spartali Stillman. Later followers of the principles of the Brotherhood included Edward Burne-Jones, William Morris and John William Waterhouse.