Ayliffe is an English surname. Notable people with this surname include:
Graham Ayliffe (1926–2017), British microbiologist John Ayliffe (1676–1732), English jurist expelled from Oxford University Royce Ayliffe (born 1956), Australian professional rugby league footballer Thomas Hamilton Ayliffe (1774–1852) and family, early settlers of South Australia See also
Ayliffe technique, a hand washing technique