Background: Iron deficiency anemia (IDA) and beta thalassaemia trait (BTT) are two of the most common causes of microcytic anemia.It is essential to differentiate between the two, so as to avoid unnecessary iron therapy which is contraindicated in beta thalassaemia .We retrospectively evaluated the reliability of various indices for differential diagnosis of microcytosis and 𝛽-TT in the same patient groups .Methods: A total of 200 patients were evaluated. We calculated 6 discrimination indices in all patients with anemia or suspected beta thalassemia. None of the subjects had combined IDA and 𝛽-TT. The patient groups were evaluated according to the Mentzer, Shine and Lal, England and Fraser,Srivastava, Green and King, Ricerca.Results: According to percentage correctly diagnosed criteria, Ricerca and Mentzer index are considered as the two best discrimination indices in differentiation between β thalassemia trait and iron deficiency anaemia .Youden’s index showed the following ranking with respect to the indices’ ability to distinguish between 𝛽-TT and IDA, Ricerca index > Mentzer index > Green and King index > Srivastava index > Shine and Lal index > England and Fraser index.Conclusions: Ricerca and Mentzer index are considered as the two best discrimination indices in differentiation between β thalassemia trait and iron deficiency anaemia.Keywords: Iron deficiency anemia (IDA) and beta thalassaemia trait (𝛽-TT)